Enzyme Substrates Spec

A derivative of BCIP. Unlike BCIP that yields a blue precipitating product, BCIP Pink produces a pink colored (l_max ~ 540 nm) precipitate. Protocol for using this product is similar to that for BCIP.

• White solid soluble in water
• Store at -20 °C and protect from light
• C₁₅H₁₇ClN₂O₄P.
• Mwt: 354.7
• [159954-33-3]

A derivative of BCIP. Unlike BCIP that yields a blue precipitating product, BCIP Red produces a red (l_max ~ 565 nm) colored precipitate.

• White solid soluble in water
• Store at -20 °C and protect from light
• C₁₅H₁₅BrClN₂O₄P.
• Mwt: 433.6
• [6769-80-8]

See #10001 for product description.

See #10001 for product description.

BCIP is the most widely used chromogenic phosphatase substrate, which forms a dark blue (l_max ~615 nm) precipitate on enzymatic hydrolysis. It is often used with the oxidant NBT (nitro blue tetrazolium chloride,#10008), which facilitates the precipitation. For your convenience, we offer a BCIP/NBT Kit (# 10003) that contains both reagents with a price lower than the combined prices of the components. Please also see our BCIP derivatives below that yield other colored precipitates.

• White solid soluble in water.
• Store at -20°C and protect from light.
• C₈H₄BrClNNa₂O₄P^.1.5H₂O
• Mwt: 397.5.
• [102185-33-1].

See #10002 for product description.

See #10002 for product description.

Similar to BCIP/Na (10001)

• Store at -20 °C and protect from light.
• C₁₅H₁₅BrClN₂O₄P.
• Mwt: 433.6.
• [6578-06-9].

This kit contains 100 mg each of BCIP Pink (10006) and the precipitation inducer NBT (10008).

This kit contains 100 mg each of BCIP Red (10004) and the oxidant NBT (10008).

This kit contains 100 mg each of BCIP sodium salt (10001) and the oxidant NBT (nitro blue tetrazolium, 10008).

The sampler kit contains 10 mg each of BCIP Na (#10001), BCIP Pink (#10006) and BCIP Red (#10004) and 250 mg of NBT (#10008). NBT forms a co-precipitant and enhances the BCIP reaction.

FDP is the most sensitive fluorogenic alkaline phosphatase substrate.¹ It is also an excellent substrate for protein tyrosine phosphatase (PTPase) and thus the substrate of choice in the high throughput screening (HTS) for PTPase inhibitor. FDP itself is colorless, but the end product of the enzymatic reaction is fluorescein, which is highly green fluorescent.

• l_ex\l_em= 490/514 nm (end product)
• e ≈ 90,000 (end product)
• Off-white solid soluble in water
• Store at -20 °C and protect from light, especially in solution
• C₂₀H₂₆N₄O₁₁P₂
• Mwt: 560.39
• [217305-49-2]

MUP is a fluorogenic phosphatase substrate that releases the blue fluorescent dye 4-methyl-7-hydroxycoumarin. MUP free acid and its salt (See MUP, Na, #10010 ) are the most widely used fluorogenic substrates for alkaline phosphatase detection.

• l_ex\l_em= 360/450 nm (end product)
• White solid soluble in water and DMSO.
• Store desiccated at -20 °C and protect from light.
• C₁₀H₉O₆P.
• Mwt: 256
• [3368-04-5].

See #10009 for product description.

See #10010 for product description.

Sodium salt of MUP. Its use is similar to that of MUP free acid (#10009)

• C₁₀H₇Na₂O₆P^.3H₂O
• Mwt: 354.2
• [22919-26-2]

See #10008 for product description.

See #10008 for product description.

NBT is an oxidant and is widely used in conjuction with BCIP or BCIP derivatives to detect phosphotase activity. NBT is also a substrate for dehydrogenase and other oxidases. Thus, it is often used to assess cell viability, proliferation and cytotoxicity, and is also frequently used in histochemistry studies. The colorless NBT is soluble in water, but forms a dark blue-colored, highly insoluble precipitate after being enzymatically reduced.

 

• White solid soluble in water and DMSO.
• Store at 4 °C.
• C₄₀H₃₀Cl₂N₁₀O₆.
• Mwt: 817.7.
• [298-83-9].

Chromogenic substrate for b-galactosidase. Similar to X-Gal, but generates green colored precipitate on enzyme hydrolysis.

• White solid soluble in water and DMSO.
• Store at -20°C and protect from light.
• C₁₅H₁₉NO₆.
• Mwt: 309.3.
• [207598-26-3]

Chromogenic substrate for b-galactosidase. Similar to X-gal, but generates a purple (l_max ~575 nm) colored precipitate on enzyme hydrolysis.

• Off-white solid soluble in water and DMSO.
• Store at -20°C and protect from light.
• C₁₄H₁₆INO₆.

Chromogenic substrate for b-galactosidase. Similar to X-gal, but generates a red or magenta (l_max ~ 565 nm) colored precipitate on enzyme hydrolysis.

• White solid soluble in DMSO
• Store at -20°C and protect from light
• C₁₄H₁₅BrClNO₆.
• Mwt: 408.6
• [93863-88-8]

Rose-b-D-Gal is a chromogenic substrate for b-galactosidase. The product is similar to X-gal, but generates an intense pink colored precipitate (l_max ~ 540 nm) on enzymatic hydrolysis. Protocol for using the material is similar to that for X-gal (See references listed below).

• White solid soluble in DMSO.
• Store at -20°C
• C₁₄H₁₆ClNO₆.

See #10009 for product description.

X-gal is the most widely used chromogenic substrate for b-galactosidase. Enzyme hydrolysis of the substrate yields a dark blue colored precipitate (l_max ~ 615nm).

• White solid soluble in DMSO.
• Store at -20°C and protect from light.
• C₁₄H₁₅BrClNO₆.
• Mwt: 408.6.
• [7240-90-6].

See #10011 for product description.

For researchers wishing to investigate the suitability of our various chromogenic b-galactosidase substrates, we offer this sampler kit, which contains 5 mg each of the following five different substrates: X-Gal (#10011), Red-b-D-Gal (#10012), Rose-b-D-Gal (#10013), Purple-b-D-Gal (#10014) and Green-b-D-Gal (#10015).

FDG is perhaps the most sensitive fluorogenic substrate for b-galactosidase. The nonfluorescent substrate is hydrolyzed step-wise by the enzyme to generate first the fluorescent monogalactoside and then the final fluorescent product fluorescein. In addition to use for gene expression detection, FDG has also been used in ELISAs, where the enzyme is linked to an antibody.

• l_ex\l_em(pH8) = 490/514 nm (end product)
• e≈90,000 (end product)
• Off-white solid soluble in water
• Store at -20°C and protect from light, especially in solution
• C₃₂H₃₂O₁₅
• Mwt: 656.59
• [17817-20-8]

Red-b-D-GlcU is similar to X-GlcU, but the product of the substrate is a red (l_max ~ 565 nm) precipitate, which is easier to detect against the background of plant cells.

• White solid soluble in water and DMSO
• Store desiccated at -20°C and protect from light
• C₂₀H₂₆BrClN₂O₇.
• Mwt: 521.8
• [144110-43-0]

Rose-b-D-GlcU is similar to X-GlcU, but the product of the substrate is a rosecolored (l_max ~ 540 nm) precipitate, which is easier to detect against the background of plant cells.

• White solid soluble in H₂O or DMSO
• Store at -20°C and protect from light
• C₂₀H₂₇ClN₂O₇
• Mwt: 412.9
• [138182-20-4]

X-GlcU is a chromogenic β-glucuronidase substrate widely used for monitoring E. coli contamination in food (such as meat, dairy products and shellfish, etc) and water, and for the detection of GUS gene expression in plants. X-GlcU also has clinical applications in the assessment of urinary tract infection by detecting the presence of E. coli. The substrate yields a dark blue (l_max ~ 615 nm) precipitate after enzymatic hydrolysis.

• White solid soluble in water or DMSO
• Store desiccated at -20°C and protect from light
• ₂₀H₂₆BrClN₂O₇.
• Mwt: 521.8
• [18656-96-7]

X-GlcU, Na is similar to X-GlcU, CHA (#10016), but it has better water solubility than the latter.

• White solid soluble in water or DMSO
• Store desiccated at -20°C and protect from light
• C₁₄H₁₂BrClNNaO₇.
• Mwt: 444.6
• [12954-41-9]

For researchers wishing to investigate the suitability of our various chromogenic b-glucuronidase substrates, we offer this sampler kit, which contains 5 mg each of the following three different substrates: X-GlcU, CHA (#10016), Red-b- D-GlcU, CHA (#10018), and Rose-b-D-GlcU, CHA(#10019).

MUG is a fluorogenic b-glucuronidase substrate that releases the blue fluorescent 4-methyl-7-hydroxycoumarin on enzymatic hydrolysis. The substrate is commonly used for identifying E. coli contamination and for detecting marker GUS gene expression in plants with high sensitivity.

• l_ex\l_em= 360/450 nm (end product)
• White solid soluble in DMSO
• Store desiccated at -20°C and protect from light
• C₁₆H₁₆O₉^.2H₂O.
• Mwt: 388.3
• [6160-80-1]

10-Acetyl-3,7-dihydroxyphenoxazine, which has been marketed under the name Amplex Red^TM reagent, is regarded as the best fluorogenic substrate for peroxidase because it is highly specific and stable. The substrate itself is nearly colorless and nonfluorescent until it is oxidized by H₂O₂ in the presence of horseradish peroxidase (HRP) to become the highly red fluorescent resorufin. High purity resorufin (#10062) is also available from Biotium to be used as a standard.

• l_ex\l_em(pH 9)= 571/585 nm (end product)
• e= 54,000 (end product)
• White to off-white solid soluble in DMSO or DMF
• Store desiccated at ?20 °C and protect from light
• C₁₄H₁₁CNO₄
• Mwt: 257.24
• [119171-73-2]

H₂DCFDA is a useful fluorogenic reagent to detect reactive oxygen intermediates in cells. On oxidation, H₂DCFDA becomes the highly green fluorescent 2',7'-dichlorofluorescein.

 

• l_ex\l_em(pH>6) = 504/529 nm (end product)
• White solid soluble in DMSO or DMF
• Store desiccated at -20°C and protect from light
• C₂₄H₁₆Cl₂O₇.
• Mwt: 487.29
• [4091-99-0]

ABTS is a very sensitive chromogenic peroxidase substrate widely used for ELISA applications. The substrate yields a water-soluble green colored product.

• Pale yellow solid soluble in water
• Store desiccated at 4°C and protect from light
• C₁₈H₂₄N₆O₆S₄.
• Mwt: 549
• [30931-67-0]

Dihydroethidium is the chemically reduced form of the commonly used DNA dye ethidium bromide. The probe is useful to detect oxidative activities in viable cells, including respiratory burst in phagocytes. Dihydroethidium itself is blue fluorescent (l_ex\l_em: 355/420nm) in cell cytoplasm while the oxidized form ethidium is red fluorescent (l_ex\l_em: 518/605 nm) upon DNA intercalation

• Off-white to light brown solid soluble in DMF or DMSO
• Store at -20°C. Under nitrogen or argon, and protect from light, especially when in solution.
• C₂₁H₂₁N₃
• Mwt: 315.
• [38483-26-0]

Dihydrorhodamine 123 is the reduced form of rhodamine 123 ( 70010), which is a commonly used fluorescent mitochondrial dye. Dihydrorhodamine 123 itself is nonfluorescent, but it readily enters most of the cells and is oxidized by oxidative species or by cellular redox systems to the fluorescent rhodamine 123 that accumulates in mitochondrial membranes.¹ Dihydrorhodamine 123 is useful for detecting reactive oxygen species including superoxide (in the presence of peroxidase or cytochrome c)^2,3 and peroxynitrite.^4,5

 

• White solid soluble in DMSO.
• Store at -20°C and protect from air and light, especially when in solution.
• C₂₀H₁₈N₂O₃.
• Mwt: 346.
• [109244-58-8]

Also see dihydrorhodamine 123 dihydrochloride ( 10056), a more stable and water soluble form of dihydrorhodamine 123.

Dihydrorhodamine 123 dihydrochloride is functionally equivalent to dihydrorhodamine 123 (#10055) but with increased stability toward air oxidation and light during storage.

• White solid soluble in DMSO
• Store at -20°C and protect from light, especially in solution
• C₂₀H₂₀Cl₂N₂O₃.
• Mwt: 419

High purity resorufin that can be used as a standard for enzyme substrates whose end product is resorufin.

• l_ex\l_em(pH9) = 563/587nm
• Dark solid soluble in DMSO
• Store at -20°C and protect from light
• C₁₂H₇NO₃
• Mwt: 213.04 N

MTT is a tetrazolium salt that is commonly used to detect reductive metabolism in cells for viability, proliferation and cytotoxicity assays. On reduction tetrazolium salts turn into deeply colored products that can be easily quantified by colorimetric measurement. MTT forms a purple formazan product that needs to be solubilized prior to quantification. MTT and XTT(#10060) are particularly useful for HTS of antiviral and anticancer compounds and for the studies of cytokine effect on cell proliferation. Please also see MTT Cell Proliferation Assay Kit in Bioscience Kit Section.

• Yellow solid soluble in water or DMSO
• C₁₈H₁₆BrN₅S
• Mwt: 414.32
• [298-93-1]

Resazurin, which is the N-oxide of the fluorescent dye resorufin and has also been under the name AlamarBlue^TM, is useful for detecting reductive activities in cells and has been widely used for measuring cell proliferation^1,2 and mitochondrial metabolic activity. Resazurin itself is nonfluorescent until it is reduced to the highly red fluorescent resorufin (#10062). Usually, NADPH or NADH is the reductant that converts resazurin to resorufin in the presence of diaphorase as the enzyme. Thus, resazurin can be used to detect NADH, NADPH, or diaphorase level. Furthermore, the resazurin/diaphorase/ NADPH system can be used to detect any biochemical or enzyme activity that is involved in a biochemical reaction generating NADH or NADPH.^3-9

Although resazurin is available from most of the chemical suppliers, a trace amount of resorufin contaminant often make the material unsuitable for bioassay applications. We supply a high grade resazurin that has a minimal background fluorescence. Please also see Resazurin Cell Proliferation Assay Kit in Bioscience Kit Section.

• l_ex\l_em(pH9) = 604nm/none
• Dark solid soluble in DMSO or H2O
• Store at -20°C and protect from light
• C₁₂H₆NNaO₄
• Mwt: 251.17

A tetrazolium derivative used widely in cell viability and proliferation testings. The material is readily reduced in viable cells to a highly water-soluble orange colored product (l= 475 nm). Applications of the reagent include studies of : antifungal susceptibility,¹ drug sensitivity of cells,² parasitic nematode viability³ and tumor cell cytotoxicity.⁴ Please also see XTT Cell Proliferation Assay Kit in the Bioscience Kit Section.

• Pale yellow solid soluble in water.
• Store desiccated at 4°C.
• C₂₂H₁₂N₇NaO₁₃S₂.
• Mwt: 675.
• [111072-31-2].

DMNPE-caged D-luciferin is a D-luciferin ester derivative which can cross cell membranes efficiently.^1,2 Once inside the cells, the ester is continuously hydrolyzed to a supply of D-luciferin. Alternatively a burst of D-luciferin is generated by UV photolysis.

• Light yellow solid soluble in DMSO.
• Store at -20°C and protect from light.
• C₂₁H₁₉N₃O₇S₂.
• Mwt: 489.52

See #10100 for product description.

See #10100 for product description.

D-Luciferin is a natural compound isolated from fireflies and is a substrate for the enzyme luciferase. Chemiluminescence at 560 nm is generated when luciferin is oxidized by an ATP-dependent process, catalyzed by luciferase. Luciferase is encoded by the luc gene, which is widely used as a reporter gene in a variety of cells. Because of the intrinsic low background of chemiluminescence technique, detection of the luc gene expression can be made at very low level. In addition, luciferin/luciferase has been used to measure 10^-15 molar quantity of ATP. White solid soluble in DMSO and slightly soluble in water.

 

• Store at -20°C and protect from light.
• C₁₁H₈N₂O₃S₂.
• Mwt: 280.
• [2591-17-5].

See #10101 for product description.

See #10101 for product description

The water-soluble form of D-luciferin. Light yellow solid soluble in water.

• Store at -20°C and protect from light.
• C₁₁H₇KN₂O₃S₂.
• Mwt: 318.4.
• [115144-35-9].

See #10102for product description.

See #10102 for product description.

The water-soluble form of D-luciferin. White solid soluble in water.

 

• White solid soluble in water.
• Store at -20°C and protect from light.
• C₁₁H₇N₂NaO₃S₂.H₂O.
• Mwt: 302.3.
• [103404-75-7]