List of Figures
Figure 1 — Jablonski Diagram Illustrating the Creation of an Excited Electronic Singlet State
Figure 2 — Excitation of a Fluorophore at Three Different Wavelengths
Figure 3 — Fluorescence Detection of Mixed Species
Figure 4 — Absorption and Fluorescence Spectral Ranges for a Variety of Important Fluorophores
Figure 1.1 — Reaction of a primary amine with an isothiocyanate
Figure 1.2 — Reaction of a primary amine with a succinimidyl ester or a tetrafluorophenyl (TFP) ester
Figure 1.3 — Reaction of a primary amine with an STP ester
Figure 1.4 — Reaction of a primary amine with a sulfonyl chloride
Figure 1.5 — Illustration of the three simple steps in the protocol for Molecular Probes' Protein Labeling Kits
Figure 1.6 — Illustration of the three simple steps in the protocol for Molecular Probes' Monoclonal Antibody Labeling Kits
Figure 1.7 — B6352; 6-((6-((biotinoyl)amino)hexanoyl)amino)hexanoic acid, sulfosuccinimidyl ester, sodium salt (biotin-XX, SSE)
Figure 1.8 — Absorption and fluorescence emission spectra of fluorescein and Alexa Fluor 488 antibody conjugates
Figure 1.9 — Photobleaching resistance of three green fluorophores, as determined by laser-scanning cytometry
Figure 1.10 — Photobleaching comparison of fluorescein phalloidin and Alexa Fluor(R) 488 phalloidin
Figure 1.11 — Bovine pulmonary artery endothelial cells (BPAEC). Alexa Fluor(R) 488 phalloidin, anti–α-tubulin mouse monoclonal antibody, Alexa Fluor(R) 546 goat anti–mouse IgG antibody.
Figure 1.12 — Comparison of pH-dependent fluorescence of green-fluorescent fluorophores
Figure 1.13 — Comparison of the relative fluorescence of Alexa Fluor 488 and FITC conjugates
Figure 1.14 — Brightness comparison of Molecular Probes' Alexa Fluor 488 dye and Cy2 dye antibody conjugates
Figure 1.15 — A30005; Alexa Fluor 488 carboxylic acid, 2,3,5,6-tetrafluorophenyl ester (Alexa Fluor 488 5-TFP)
Figure 1.16 — Stability of the tetrafluorophenyl (TFP) and succinimidyl (NHS) esters at basic pH
Figure 1.17 — Absorption spectra of our intermediate-wavelength light–absorbing Alexa Fluor dyes
Figure 1.18 — Comparison of the absorption and fluorescence emission spectra of the Alexa Fluor 555 and Cy3 dyes
Figure 1.19 — Neuronal cells in a 22-hour zebrafish embryo identified with anti–HuC/HuD mouse monoclonal antibody.
Figure 1.20 — Photobleaching profiles of the Alexa Fluor 555 and Cy3 dyes
Figure 1.21 — Comparison of the relative fluorescence of Alexa Fluor 594 and Texas Red-X goat anti–mouse IgG antibody F(ab'){2} fragment conjugates
Figure 1.22 — Flow cytometry comparison of the brightness of the Alexa Fluor 555 goat anti–mouse IgG antibody with commercially available Cy3 goat anti–mouse IgG antibody conjugates
Figure 1.23 — Comparison of the fluorescence emission of Alexa Fluor 546 and Cy3 antibody conjugates
Figure 1.24 — Absorption spectra of our long-wavelength light–absorbing Alexa Fluor dyes
Figure 1.25 — Comparison of the fluorescence spectra of the Alexa Fluor 647 and Cy5 dyes
Figure 1.26 — Comparison of the fluorescence spectra of the unconjugated Alexa Fluor 680 and Cy5.5 dyes
Figure 1.27 — Comparison of the fluorescence emission spectra of the Alexa Fluor 750 and Cy7 dyes
Figure 1.28 — Photobleaching resistance of five red fluorophores, as determined by laser-scanning cytometry
Figure 1.29 — Comparison of the relative fluorescence of goat anti–rabbit IgG antibody conjugates of the Alexa Fluor 555 and Cy3 dyes
Figure 1.30 — Comparison of the brightness of Alexa Fluor 647 and Cy5 dye conjugates
Figure 1.31 — Comparison of the brightness of Alexa Fluor 647 conjugates and Cy5 conjugates
Figure 1.32 — Flow cytometry comparison of the brightness of the Alexa Fluor 647 goat anti–mouse IgG antibody conjugate with commercially available Cy5 goat anti–mouse IgG antibody conjugates
Figure 1.33 — Comparison of the absorption spectra of Alexa Fluor 647 and Cy5 dye conjugates
Figure 1.34 — Absorption spectra of our short-wavelength light–absorbing Alexa Fluor dyes
Figure 1.35 — Bovine pulmonary artery endothelial (BPAE) cell. Anti–bovine α-tubulin mouse monoclonal antibody and Alexa Fluor(R) 430 goat anti–mouse IgG antibody.
Figure 1.36 — Bovine pulmonary artery endothelial (BPAE) cells labeled with mouse monoclonal anti–α-tubulin antibody and detected using TSA Kit #7 with the HRP conjugate of goat anti–mouse IgG antibody and Alexa Fluor(R) 350 tyramide.
Figure 1.37 — A zebrafish retina cryosection labeled with the mouse monoclonal antibody FRet 6 and detected using TSA Kit #9 with the HRP conjugate of goat anti–mouse IgG antibody and Alexa Fluor(R) 488 tyramide.
Figure 1.38 — A zebrafish retina cryosection labeled with the mouse monoclonal antibody FRet 43 and detected using TSA Kit #9 with the HRP conjugate of goat anti–mouse IgG antibody and Alexa Fluor(R) 488 tyramide.
Figure 1.39 — Bovine pulmonary artery endothelial cells (BPAEC) labeled with anti–OxPhos Complex IV subunit I antibody and detected using TSA Kit #4 with the HRP conjugate of goat anti–mouse IgG antibody and Alexa Fluor(R) 568 tyramide.
Figure 1.40 — Bovine pulmonary artery endothelial cells (BPAEC) labeled with anti–OxPhos Complex IV subunit I (human) antibody and detected using TSA Kit #6 with the HRP conjugate of goat anti–mouse IgG antibody and Alexa Fluor(R) 647 tyramide.
Figure 1.41 — Normalized fluorescence emission spectra of seven BODIPY fluorophores
Figure 1.42 — Ring-numbering system of the BODIPY fluorophores
Figure 1.43 — Emission spectra of fluorescein, TMR and TR conjugates
Figure 1.44 — BODIPY FL/MeOH
Figure 1.45 — NIH 3T3 cells. MitoTracker(R) CMXRos, BODIPY(R) FL phallacidin and POPO™-1.
Figure 1.46 — Comparison of photostability of green-fluorescent antibody conjugates
Figure 1.47 — Demonstration of single-photon and two-photon excitation.
Figure 1.48 — D6141; DISCONTINUED N-(4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene-3-propionyl)cysteic acid, succinimidyl ester, triethylammonium salt (DISCONTINUED BODIPY FL, CASE)
Figure 1.49 — Microtubules of a sea urchin embryo visualized with a BODIPY(R) FL goat anti–rabbit IgG secondary antibody.
Figure 1.50 — Mouse fibroblasts. BODIPY(R) TR-X phalloidin, DAPI, BODIPY(R) FL goat anti–rabbit IgG antibody.
Figure 1.51 — Live bovine pulmonary artery endothelial cells (BPAEC) labeled with LysoTracker(R) Red and Hoechst 33342.
Figure 1.52 — Fluorescein/pH 9.0
Figure 1.53 — Photobleaching profiles of cells stained with Alexa Fluor 488 or fluorescein conjugates
Figure 1.54 — Comparison of relative fluorescence of green-fluorescent antibody conjugates
Figure 1.55 — F143; fluorescein-5-isothiocyanate (FITC 'Isomer I')
Figure 1.56 — Proteobacterial symbionts. Fluorescein-5-isothiocyanate and Texas Red(R) sulfonyl chloride.
Figure 1.57 — F6106; 6-(fluorescein-5-carboxamido)hexanoic acid, succinimidyl ester (5-SFX)
Figure 1.58 — F6130; fluorescein-5-EX, succinimidyl ester
Figure 1.59 — C20050; 5-carboxyfluorescein-bis-(5-carboxymethoxy-2-nitrobenzyl) ether, beta-alanine-carboxamide, succinimidyl ester (CMNB-caged carboxyfluorescein, SE)
Figure 1.60 — Oregon Green 488 goat anti–mouse IgG antibody/pH 8.0
Figure 1.61 — CRE BAG 2 fibroblasts. Oregon Green(R) 514 phalloidin and fluorescein phalloidin.
Figure 1.62 — O6146; Oregon Green 488 carboxylic acid
Figure 1.63 — O6138; Oregon Green 514 carboxylic acid
Figure 1.64 — O6185; Oregon Green 488-X, succinimidyl ester
Figure 1.65 — Normalized emission spectra of 5-FAM SE, 6-TET SE, 6-JOE SE, and 6-HEX SE
Figure 1.66 — Structures of 6-JOE, SE, 6-HEX SE and 6-TET SE
Figure 1.67 — C6166; DISCONTINUED 5-carboxy-2',4',5',7'-tetrabromosulfonefluorescein, succinimidyl ester, bis-(diisopropylethylammonium) salt
Figure 1.68 — Conjugation of Rhodamine Green TFA SE to an amine
Figure 1.69 — The julolidine ring structure of X-rhodamine, sulforhodamine 101 and Texas Red dyes
Figure 1.70 — Normalized absorption spectra of the QSY 7, QSY 9, QSY 21 and QSY 35 dyes
Figure 1.71 — Effect of protein conjugation on the absorption spectrum of tetramethylrhodamine
Figure 1.72 — T6105; 6-(tetramethylrhodamine-5-(and-6)-carboxamido)hexanoic acid, succinimidyl ester (5(6)-TAMRA-X, SE)
Figure 1.73 — L20; Lissamine rhodamine B sulfonyl chloride
Figure 1.74 — Emission spectra of goat anti–mouse IgG antibody conjugates
Figure 1.75 — R6160; Rhodamine Red-X, succinimidyl ester
Figure 1.76 — Comparison of the relative fluorescence of Rhodamine Red-X and Lissamine rhodamine B conjugates
Figure 1.77 — X491; X-rhodamine-5-(and-6)-isothiocyanate (5(6)-XRITC)
Figure 1.78 — 5-ROX/pH 7.0
Figure 1.79 — C1309; 5-(and-6)-carboxy-X-rhodamine, succinimidyl ester (5(6)-ROX, SE)
Figure 1.80 — Mouse fibroblasts. BODIPY(R) FL phallacidin, Texas Red(R) goat anti–mouse IgG (H+L) antibody and DAPI.
Figure 1.81 — T353; Texas Red sulfonyl chloride
Figure 1.82 — T20175; Texas Red-X, succinimidyl ester
Figure 1.83 — Comparison of the relative fluorescence of Texas Red-X and Texas Red sulfonyl chloride conjugates
Figure 1.84 — C653; 5-(and-6)-carboxynaphthofluorescein, succinimidyl ester
Figure 1.85 — Carboxynaphthofluorescein/pH 10.0
Figure 1.86 — Q10193; QSY 7 carboxylic acid, succinimidyl ester
Figure 1.87 — Malachite green isothiocyanate/MeCN
Figure 1.88 — M689; malachite green isothiocyanate
Figure 1.89 — Reaction of NANOGOLD mono(sulfosuccinimidyl ester) with a primary amine
Figure 1.90 — A10168; Alexa Fluor 350 carboxylic acid, succinimidyl ester
Figure 1.91 — A6118; 6-((7-amino-4-methylcoumarin-3-acetyl)amino)hexanoic acid, succinimidyl ester (AMCA-X, SE)
Figure 1.92 — Alexa Fluor 350 goat anti–mouse IgG antibody/pH 8.0
Figure 1.93 — Microtubules of fixed bovine pulmonary artery endothelial cells (BPAEC). Anti–bovine α-tubulin antibody and Alexa Fluor(R) 350 goat anti–mouse IgG antibody.
Figure 1.94 — A10169; Alexa Fluor 430 carboxylic acid, succinimidyl ester
Figure 1.95 — Comparison of the pH-dependent fluorescence changes produced by attachment of fluorine atoms to a hydroxycoumarin
Figure 1.96 — M10165; Marina Blue succinimidyl ester
Figure 1.97 — P10163; Pacific Blue succinimidyl ester
Figure 1.98 — A zebrafish retina cryosection visualized using TSA Kit #10 and the SYTOX(R) Orange nucleic acid stain.
Figure 1.99 — C2284; Cascade Blue acetyl azide, trisodium salt
Figure 1.100 — Emission spectra of Cascade Blue dye, aminomethylcoumarin and fluorescein
Figure 1.101 — Cascade Blue BSA/pH 7.0
Figure 1.102 — A30000; Alexa Fluor 405 carboxylic acid, succinimidyl ester
Figure 1.103 — P6114; N-(1-pyrenebutanoyl)cysteic acid, succinimidyl ester, potassium salt
Figure 1.104 — D6104; 6-((5-dimethylaminonaphthalene-1-sulfonyl)amino)hexanoic acid, succinimidyl ester (dansyl-X, SE)
Figure 1.105 — B30250; bimane mercaptoacetic acid (carboxymethylthiobimane)
Figure 1.106 — S6110; 1-(3-(succinimidyloxycarbonyl)benzyl)-4-(5-(4-methoxyphenyl)oxazol-2-yl)pyridinium bromide (PyMPO, SE)
Figure 1.107 — C10164; Cascade Yellow succinimidyl ester
Figure 1.108 — Emission spectra of Pacific Blue and Cascade Yellow antibody conjugates
Figure 1.109 — D12800; Dapoxyl sulfonic acid, sodium salt
Figure 1.110 — Dapoxyl (2-aminoethyl)sulfonamide/MeOH
Figure 1.111 — Emission spectra of a Dapoxyl dye in five different solvents
Figure 1.112 — Bovine pulmonary artery endothelial cells (BPAEC). LysoTracker(R) Blue-White DPX and MitoTracker(R) Red CMXRos.
Figure 1.113 — Bovine pulmonary artery endothelial cells (BPAEC). ER-Tracker™ Blue-White DPX.
Figure 1.114 — FluoSpheres(R) fluorescent microspheres.
Figure 1.115 — Fluorogenic amine-derivitization reaction of fluorescamine
Figure 1.116 — Fluorogenic amine-derivitization reaction of o-phthaldialdehyde (OPA)
Figure 1.117 — Fluorogenic amine-derivitization reaction of naphthalene-2,3-dicarboxaldehyde (NDA)
Figure 1.118 — Fluorogenic amine-derivitization reaction of CBQCA
Figure 1.119 — C20260; 4-chloro-7-nitrobenz-2-oxa-1,3-diazole (NBD chloride; 4-chloro-7-nitrobenzofurazan)
Figure 1.120 — D1537; DISCONTINUED 4-dimethylaminoazobenzene-4'-sulfonyl chloride (DISCONTINUED dabsyl chloride)
Figure 1.121 — Normalized absorption spectra of the succinimidyl esters of dabcyl and QSY 35 dyes
Figure 1.122 — D2245; 4-((4-(dimethylamino)phenyl)azo)benzoic acid, succinimidyl ester (dabcyl, SE)
Figure 1.123 — Q20133; QSY 35 acetic acid, succinimidyl ester
Figure 2.1 — Structural comparison of the reducing agents DTT and TCEP
Figure 2.2 — Reaction of a thiol with an alkyl halide
Figure 2.3 — Reaction of a thiol with a maleimide
Figure 2.4 — Reaction of a thiol with a symmetric disulfide
Figure 2.5 — A10254; Alexa Fluor 488 C{5}-maleimide
Figure 2.6 — Reaction of an Hg-Link phenylmercury compound with a nitrosylated thiol
Figure 2.7 — Comparison of the fluorophore orientation relative to the reactive moiety of two spectrally similar thiol-reactive BODIPY dyes
Figure 2.8 — Reaction of intramolecularly quenched BODIPY FL L-cystine with a thiol
Figure 2.9 — Reaction of a TS-Link reagent with a thiol, followed by removal of the label with a reducing agent
Figure 2.10 — F150; fluorescein-5-maleimide
Figure 2.11 — B1355; 5-(bromomethyl)fluorescein
Figure 2.12 — O6034; Oregon Green 488 maleimide
Figure 2.13 — T6027; tetramethylrhodamine-5-maleimide
Figure 2.14 — T6028; tetramethylrhodamine-6-maleimide
Figure 2.15 — R6029; Rhodamine Red C{2}-maleimide
Figure 2.16 — T6009; Texas Red C{5}-bromoacetamide
Figure 2.17 — T6008; Texas Red C{2}-maleimide
Figure 2.18 — M6026; 1-(2-maleimidylethyl)-4-(5-(4-methoxyphenyl)oxazol-2-yl)pyridinium methanesulfonate (PyMPO maleimide)
Figure 2.19 — D2004; N,N'-dimethyl-N-(iodoacetyl)-N'-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)ethylenediamine (IANBD amide)
Figure 2.20 — Lucifer yellow CH/H{2}O
Figure 2.21 — Q10257; QSY 7 C{5}-maleimide
Figure 2.22 — Reaction of NANOGOLD monomaleimide with a thiol
Figure 2.23 — Labeling of IgG molecules with NANOGOLD monomaleimide.
Figure 2.24 — D346; 7-diethylamino-3-(4'-maleimidylphenyl)-4-methylcoumarin (CPM)
Figure 2.25 — D10253; 7-diethylamino-3-((((2-maleimidyl)ethyl)amino)carbonyl)coumarin (MDCC)
Figure 2.26 — P28; N-(1-pyrene)maleimide
Figure 2.27 — A433; 6-acryloyl-2-dimethylaminonaphthalene (acrylodan)
Figure 2.28 — Fluorescence emission spectra of the 2-mercaptoethanol adduct of badan in six solvents
Figure 2.29 — I7; 2-(4'-(iodoacetamido)anilino)naphthalene-6-sulfonic acid, sodium salt (IAANS)
Figure 2.30 — D10300; Dapoxyl (2-bromoacetamidoethyl)sulfonamide
Figure 2.31 — M1378; monobromobimane (mBBr)
Figure 2.32 — M1380; DISCONTINUED monobromotrimethylammoniobimane bromide (DISCONTINUED qBBr)
Figure 2.33 — D1379; dibromobimane (bBBr)
Figure 2.34 — A484; 4-acetamido-4'-((iodoacetyl)amino)stilbene-2,2'-disulfonic acid, disodium salt
Figure 2.35 — A485; 4-acetamido-4'-maleimidylstilbene-2,2'-disulfonic acid, disodium salt
Figure 3.1 — Oxidation of an N-terminal serine residue to an aldehyde
Figure 3.2 — C20260; 4-chloro-7-nitrobenz-2-oxa-1,3-diazole (NBD chloride; 4-chloro-7-nitrobenzofurazan)
Figure 3.3 — Reaction scheme for the conversion of tyrosine to o-aminotyrosine
Figure 3.4 — Reaction of N-methylisatoic anhydride with an alcohol
Figure 3.5 — D2281; m-dansylaminophenylboronic acid
Figure 3.6 — Reaction of m-dansylaminophenylboronic acid
Figure 3.7 — Derivatization of an alcohol using the diacetate of fluorescein-5-carbonyl azide
Figure 3.8 — Oxidation of the terminal galactose residue of a glycoprotein, glycolipid or polysaccharide
Figure 3.9 — L20492; DISCONTINUED N-levulinoyl-D-mannosamine (DISCONTINUED ManLev)
Figure 3.10 — A10550; N-(aminooxyacetyl)-N'-(D-biotinoyl) hydrazine, trifluoroacetic acid salt (ARP)
Figure 3.11 — Jurkat cells. ManLev tetraacetate, ARP and Alexa Fluor(R) 488 streptavidin.
Figure 3.12 — A6257; 8-aminopyrene-1,3,6-trisulfonic acid, trisodium salt (APTS)
Figure 3.13 — Structures of a hydrazide, a semicarbazide and a carbohydrazide
Figure 3.14 — Modifying aldehydes and ketones with hydrazine derivatives
Figure 3.15 — F121; fluorescein-5-thiosemicarbazide
Figure 3.16 — APR motor neuron of a larval moth, Manduca sexta. Alexa Fluor(R) 488 hydrazide, sodium salt.
Figure 3.17 — CRE BAG 2 cells. BODIPY(R) FL hydrazide, Influx™ pinocytic cell-loading reagent and LysoTracker(R) Red DND-99.
Figure 3.18 — Reaction scheme illustrating the principle of ketone and aldehyde detection by NBD methylhydrazine
Figure 3.19 — Lipopolysaccharide staining with the Pro-Q Emerald 300 Lipopolysaccharide Gel Stain Kit
Figure 3.20 — Characterization of lipopolysaccharides using the Pro-Q Emerald 300 Lipopolysaccharide Gel Stain Kit
Figure 3.21 — Linearity of the Pro-Q Emerald 300 stain for lipopolysaccharide (LPS) detection
Figure 3.22 — Modifying aldehydes and ketones with amine derivatives
Figure 3.23 — Conversion of a carboxylic acid group into an aliphatic amine
Figure 3.24 — Stabilization of an unstable O-acylisourea intermediate in a carbodiimide-mediated modification of a carboxylic acid with a primary amine
Figure 3.25 — Q10464; QSY 7 amine, hydrochloride
Figure 3.26 — Transglutaminase-mediated labeling of a protein using dansyl cadaverine
Figure 3.27 — Carbodiimide modification of a carboxylic acid group in a protein
Figure 3.28 — N2461; 2-(2,3-naphthalimino)ethyl trifluoromethanesulfonate
Figure 3.29 — Reaction scheme illustrating esterification of a carboxylic acid with STP
Figure 4.1 — Comparison of the structures of D-biotin and D-desthiobiotin
Figure 4.2 — Bovine pulmonary artery endothelial cells. Anti–α-tubulin mouse monoclonal 236-10501, Alexa Fluor(R) 647 goat anti–mouse IgG antibody, Alexa Fluor(R) 488 streptavidin and DAPI.
Figure 4.3 — ELISA-type assay comparing the binding capacity of biotinylated BSA and goat anti–mouse IgG
Figure 4.4 — B2604; biotin-X 2,4-dinitrophenyl-X-L-lysine, succinimidyl ester (DNP-X-biocytin-X, SE)
Figure 4.5 — Identification of cell-surface proteins in Jurkat cells labeled with the FluoReporter Cell-Surface Biotinylation Kit
Figure 4.6 — B11790; DISCONTINUED biotin-X nitrilotriacetic acid, tripotassium salt (DISCONTINUED biotin-X NTA)
Figure 4.7 — Nucleophilic attack of serine on the carbonyl group of biotin-X, SSE
Figure 4.8 — N6356; norbiotinamine, hydrochloride
Figure 4.9 — B1370; 5-((N-(5-(N-(6-(biotinoyl)amino)hexanoyl)amino)pentyl)thioureidyl)fluorescein (fluorescein biotin)
Figure 4.10 — B10570; biotin-4-fluorescein
Figure 4.11 — Quantitation of biotin-binding sites with biotin-4-fluorescein
Figure 4.12 — Motor neuron in a three-day-old chick embryo labeled with lysine-fixable biotinylated dextran
Figure 5.1 — Schematic illustration of the heterobifunctional crosslinker succinimidyl acetylthioacetate
Figure 5.2 — Confocal linescan image of calcium "puffs" in a Xenopus oocyte.
Figure 5.3 — T2556; tris-(2-carboxyethyl)phosphine, hydrochloride (TCEP)
Figure 5.4 — SPDP derivatization reactions
Figure 5.5 — Derivatization reactions with TS-Link TFP-X thiosulfate
Figure 5.6 — Two-step reaction sequence for crosslinking biomolecules using SMCC
Figure 5.7 — Chemical basis for thiol detection using the Thiol and Sulfide Quantitation Kit
Figure 5.8 — B10621; bis-((N-iodoacetyl)piperazinyl)sulfonerhodamine
Figure 5.9 — D10620; DISCONTINUED 4,4-difluoro-3,5-di(iodoacetamidomethyl)-4-bora-3a,4a-diaza-s-indacene (DISCONTINUED BODIPY FL bis-(methyleneiodoacetamide))
Figure 5.10 — A685; 5-((2-(and-3)-S-(acetylmercapto)succinoyl)amino)fluorescein (SAMSA fluorescein)
Figure 5.11 — M1618; DISCONTINUED N-((4-maleimidylmethyl)cyclohexane-1-carbonyl)-1,2-dihexadecanoyl-sn-glycero-3-phosphoethanolamine, triethylammonium salt (DISCONTINUED MMCC DHPE)
Figure 5.12 — Photoreactive crosslinking reaction of a simple aryl azide
Figure 5.13 — Photoreactive crosslinking reaction of a fluorinated aryl azide
Figure 5.14 — Photoreactive crosslinking reaction of a benzophenone derivative
Figure 5.15 — P6317; N-((2-pyridyldithio)ethyl)-4-azidosalicylamide (PEAS; AET)
Figure 5.16 — Schematic showing attachment of an amine-modified oligonucleotide to a surface using a photoreactive crosslinking reagent
Figure 5.17 — B22358; 2'-(or-3')-O-(4-benzoylbenzoyl)adenosine 5'-triphosphate, tris(triethylammonium) salt (BzBzATP)
Figure 5.18 — A1048; adenosine 5'-triphosphate, P{3}-(1-(2-nitrophenyl)ethyl) ester, disodium salt (NPE-caged ATP)
Figure 5.19 — Spectral comparison of the caged Ca{2+} reagents NP-EGTA and DMNP-EDTA
Figure 5.20 — D3034; diazo-2, tetrapotassium salt
Figure 5.21 — C7619; DISCONTINUED cyclic adenosine 5'-diphosphate ribose (DISCONTINUED cADP-ribose)
Figure 5.22 — C7074; cyclic adenosine 5'-diphosphate ribose, 1-(1-(2-nitrophenyl)ethyl) ester (NPE-caged cADP-ribose)
Figure 5.23 — CNB-caged L-glutamic acid
Figure 5.24 — Zebrafish embryo. DMNB-caged fluorescein 10,000 MW dextran.
Figure 5.25 — Caging of carboxylic acids using the hydrazone precursor of DMNPE
Figure 6.1 — Simultaneous detection of three gene targets in a whole-mount Drosophila embryo by fluorescence in situ hybridization.
Figure 6.2 — Zebrafish retina. ELF(R) 97 Immunohistochemistry Kit, tetramethylrhodamine wheat germ agglutinin and Hoechst 33342
Figure 6.3 — Luminescent Constellation™ microspheres for imaging.
Figure 6.4 — Schematic diagram of primary and secondary detection reagents
Figure 6.5 — Schematic representation of TSA detection methods applied to immunolabeling of an antigen
Figure 6.6 — Coupling of Alexa Fluor 488 tyramide to protein tyrosine side chains via peroxidase-mediated formation of an O,O'-dityrosine adduct
Figure 6.7 — HeLa cells detected with Alexa Fluor 546 tyramide
Figure 6.8 — Tyramide signal amplification of immunofluorescent staining in mouse brain sections.
Figure 6.9 — Nuclear and nonnuclear incorporation of 5-bromo-2'-deoxyuridine in live cells.
Figure 6.10 — Detection of epidermal growth factor (EGF) receptors directly or with signal amplification
Figure 6.11 — Enhancement of estrogen receptor detection sensitivity by tyramide signal amplification
Figure 6.12 — Zebrafish retina. TSA Kit #2, Alexa Fluor(R) 350 wheat germ agglutinin conjugate and TOTO(R)-3 nucleic acid stain.
Figure 6.13 — In situ hybridization of α-satellite probes to human chromosomes 1, 15 and 17 detected by tyramide signal amplification.
Figure 6.14 — Digital image analysis comparison of in situ–hybridized biotinylated alpha-satellite probes
Figure 6.15 — Principle of the enzyme-mediated formation of the ELF 97 alcohol precipitate
Figure 6.16 — Photostability comparison for tubulin preparations labeled with ELF 97 alcohol or fluorescein
Figure 6.17 — Fluorescence excitation and emission spectra of the ELF 97 alcohol precipitate
Figure 6.18 — ELF 97 photostability under intense UV illumination with a confocal laser-scanning microscope
Figure 6.19 — HeLa cell nuclei. Texas Red(R)-X streptavidin, biotin-XX goat anti–mouse IgG antibody, ELF(R) 97 Cytological Labeling Kit and Hoechst 33258
Figure 6.20 — Osteoblast cells in adult zebrafish head cryosection. ELF(R) 97 Endogenous Phosphatase Detection Kit, Texas Red(R)-X wheat germ agglutinin and Hoechst 33342 nucleic acid stain.
Figure 6.21 — Schematic diagram of the methods employed in our ELF 97 Kits
Figure 6.22 — Prostate carcinoma. ELF(R) 97 mRNA In Situ Hybridization Kit
Figure 6.23 — Mouse fibroblasts. Paclitaxel, biotin-XX goat anti–mouse IgG antibody and ELF(R) 97 Cytological Labeling Kit
Figure 6.24 — Bovine pulmonary artery endothelial cells (BPAEC). Paclitaxel, biotin-XX goat anti–mouse IgG and ELF(R) 97 Cytological Labeling Kit
Figure 6.25 — Bovine pulmonary artery endothelial cells. Biotin-XX phalloidin, ELF(R) Cytological Labeling Kit
Figure 6.26 — Cellular targets developed for visualization with the reagents in our ELF 97 Cytological Labeling Kits
Figure 6.27 — Zebrafish retina. ELF(R) 97 Immunohistochemistry Kit, Hoechst 33342 and tetramethylrhodamine wheat germ agglutinin.
Figure 6.28 — Adult zebrafish intestine. ELF(R) 97 Endogenous Phosphatase Detection Kit.
Figure 6.29 — Endogenous alkaline phosphatase activity of osteosarcoma cells. ELF(R) 97 Endogenous Phosphatase Detection Kit and Hoechst 33342.
Figure 6.30 — Adult zebrafish kidney. ELF(R) 97 Endogenous Phosphatase Detection Kit and propidium iodide.
Figure 6.31 — A comparison of the photobleaching rates of APC and Cy5 conjugates
Figure 6.32 — Absorption spectra for B-PE, R-PE and APC
Figure 6.33 — Emission spectra for B-PE, R-PE and APC
Figure 6.34 — Fluorescence emission spectra of Alexa Fluor dye–conjugates of R-phycoerythrin
Figure 6.35 — Simultaneous detection of three cell surface markers using an Alexa Fluor 610–R-phycoerythrin tandem conjugate, Alexa Fluor 488 dye and R-phycoerythrin labels
Figure 6.36 — Simultaneous detection of three cell surface markers using an Alexa Fluor 647–R-phycoerythrin tandem conjugate, Alexa Fluor 488 dye and R-phycoerythrin labels
Figure 6.37 — Fluorescence emission spectra of allophycocyanin and long-wavelength Alexa Fluor dye conjugates of allophycocyanin
Figure 6.38 — Fluorescence emission spectra of Alexa Fluor 647–R-phycoerythrin streptavidin and Cy5–R-phycoerythrin streptavidin tandem conjugates
Figure 6.39 — Emission spectra of Alexa Fluor 610–R-phycoerythrin and Texas Red–R-phycoerythrin tandem conjugates
Figure 6.40 — Comparison of immunofluorescent staining by R-phycoerythrin–dye tandem conjugates
Figure 6.41 — Analytical size-exclusion chromatograms of free streptavidin and streptavidin, R-phycoerythrin conjugate
Figure 6.42 — R-phycoerythrin used to detect DNA on a microarray
Figure 6.43 — FluoSpheres(R) fluorescent microspheres.
Figure 6.44 — Positively charged nylon membrane. TransFluoSpheres(R) fluorescent microspheres.
Figure 6.45 — Emission spectra of FluoSpheres beads
Figure 6.46 — PS-Speck™ microsphere used to demonstrate a point-spread function of a microscope's optics.
Figure 6.47 — Fluorescence excitation and emission maxima of the FluoSpheres europium luminescent microspheres
Figure 6.48 — Luminescence excitation and emission spectra of the FluoSpheres platinum luminescent microspheres
Figure 6.49 — Schematic diagram of the large Stokes shifts exhibited by our TransFluoSpheres beads
Figure 6.50 — Fluorescence emission spectra of our 488 nm light–excitable TransFluoSpheres beads
Figure 7.1 — Comparison of the brightness of Alexa Fluor 647 conjugates and Cy5 conjugates
Figure 7.2 — Alexa Fluor 350 goat anti–mouse IgG antibody/pH 8.0
Figure 7.3 — Alexa Fluor 405 goat anti–mouse IgG antibody/pH 7.2
Figure 7.4 — Alexa Fluor 430 goat anti–mouse IgG antibody/pH 7.2
Figure 7.5 — Alexa Fluor 488 goat anti–mouse IgG antibody/pH 8.0
Figure 7.6 — Alexa Fluor 500 goat anti–mouse IgG antibody/pH 7.2
Figure 7.7 — Alexa Fluor 514 goat anti–mouse IgG antibody/pH 8.0
Figure 7.8 — Alexa Fluor 532 goat anti–mouse IgG antibody/pH 7.2
Figure 7.9 — Alexa Fluor 546 goat anti–mouse IgG antibody/pH 7.2
Figure 7.10 — Alexa Fluor 555 goat anti–mouse IgG antibody/pH 7.2
Figure 7.11 — Alexa Fluor 568 goat anti–mouse IgG antibody/pH 7.2
Figure 7.12 — Alexa Fluor 594 goat anti–mouse IgG antibody/pH 7.2
Figure 7.13 — Alexa Fluor 610 goat anti–mouse IgG antibody/pH 7.2
Figure 7.14 — Alexa Fluor 633 goat anti–mouse IgG antibody/pH 7.2
Figure 7.15 — Alexa Fluor 635 goat anti–mouse IgG antibody/pH 7.2
Figure 7.16 — Alexa Fluor 647 goat anti–mouse IgG antibody/pH 7.2
Figure 7.17 — Alexa Fluor 660 goat anti–mouse IgG antibody/pH 7.2
Figure 7.18 — Alexa Fluor 680 goat anti–mouse IgG antibody/pH 7.2
Figure 7.19 — Alexa Fluor 700 goat anti–mouse IgG antibody/pH 7.2
Figure 7.20 — Alexa Fluor 750 goat anti–mouse IgG antibody/pH 7.2
Figure 7.21 — Oregon Green 488 goat anti–mouse IgG antibody/pH 8.0
Figure 7.22 — Oregon Green 514 goat anti–mouse IgG antibody/pH 8.0
Figure 7.23 — Comparison of the photostability of immunofluorescent staining by Oregon Green 514 or fluorescein goat anti–mouse IgG antibodies
Figure 7.24 — BODIPY FL goat anti–mouse IgG antibody/pH 7.2
Figure 7.25 — Rhodamine Red-X goat anti–mouse IgG antibody/pH 8.0
Figure 7.26 — Texas Red-X goat anti–mouse IgG antibody/pH 7.2
Figure 7.27 — Fluorescein goat anti–mouse IgG antibody/pH 8.0
Figure 7.28 — R-phycoerythrin/pH 7.5
Figure 7.29 — Comparison of the fluorescence spectra of the unconjugated Alexa Fluor 680 and Cy5.5 dyes
Figure 7.30 — Allophycocyanin/pH 7.5
Figure 7.31 — Comparison of the relative fluorescence of AMCA streptavidin and Alexa Fluor 350 streptavidin
Figure 7.32 — Histograms showing fluorescence per fluorophore for fluorescein and Cascade Blue conjugates
Figure 7.33 — Marina Blue goat anti–mouse IgG antibody/pH 8.0
Figure 7.34 — Pacific Blue goat anti–mouse IgG antibody/pH 8.0
Figure 7.35 — Cascade Yellow goat anti–mouse IgG antibody/pH 8.0
Figure 7.36 — Brightness comparison of Molecular Probes' Alexa Fluor 488 dye and Cy2 dye antibody conjugates
Figure 7.37 — Flow cytometry comparison of the brightness of the Alexa Fluor 555 goat anti–mouse IgG antibody with commercially available Cy3 goat anti–mouse IgG antibody conjugates
Figure 7.38 — Flow cytometry comparison of the brightness of the Alexa Fluor 647 goat anti–mouse IgG antibody conjugate with commercially available Cy5 goat anti–mouse IgG antibody conjugates
Figure 7.39 — Bovine pulmonary artery endothelial cells (BPAEC). Anti–α-tubulin antibody and Alexa Fluor(R) 488, Alexa Fluor(R) 546 and Alexa Fluor(R) 594 goat anti–mouse IgG antibodies.
Figure 7.40 — Apple leaf cell walls. Alexa Fluor(R) 488 goat anti–rabbit IgG antibody conjugate.
Figure 7.41 — Developing Drosophila melanogaster embryo. Alexa Fluor(R) 488 goat anti–mouse IgG antibody conjugate, Alexa Fluor(R) 488 goat anti–rabbit IgG antibody conjugate and Hoechst 33342.
Figure 7.42 — Maize leaf section. Alexa Fluor(R) 488 goat anti–rabbit IgG antibody.
Figure 7.43 — Peripheral nervous system of a Drosophila melanogaster embryo. Alexa Fluor(R) 488 rabbit anti–mouse IgG antibody, Alexa Fluor(R) 594 goat anti–rabbit IgG antibody and DAPI.
Figure 7.44 — Reduced background staining afforded by Image-iT(R) FX signal enhancer.
Figure 7.45 — Increased label specificity and resolution provided by Image-iT(R) FX signal enhancer.
Figure 7.46 — Demonstration of the amplification obtained with the Alexa Fluor 488 Signal Amplification Kit for Fluorescein- and Oregon Green Dye–Conjugated Probes
Figure 7.47 — Example flow cytometry results obtained using the Alexa Fluor 488 Sign | 
Product Highlights
