Novex™ AP Chromogenic Substrate (BCIP/ NBT) - FAQs

Invitrogen AP发光底物（BCIP/NBT）和Invitrogen HRP发光底物（TMB）是即用型的单一成分底物，可在免疫印迹膜或点印迹膜上分别对碱性磷酸酶（AP）或辣根过氧化酶（HRP）进行灵敏的免疫检测。AP底物的灵敏度在100 pg范围内，HRP底物在1 ng范围内。
Invitrogen AP显色底物由5-溴-4-氯-3-吲哚基-1-磷酸盐（BCIP）和硝基四氮唑蓝（NBT）的即用型溶液组成，与碱性磷酸酶反应可形成黑紫色沉淀。该底物可作为独立商品（货号WP20001）购买，也是WesternBreeze显色法检测试剂盒的一个组分。
Invitrogen HRP显色底物由3,3’,5,5’-四甲基联苯胺（TMB）的即用型无毒溶液组成，与辣根过氧化物酶反应可形成蓝色沉淀。该底物可作为独立产品（货号WP20004）购买。

*注意：使用不同的抗原和抗体进行检测，灵敏度也不同。

可以，您可单独购买显色底物（货号WP20001）。它会是紫色的显色沉淀物。

Western blotting is based on the separation of proteins by their size on a gel. However, migration of proteins through the gel matrix is also affected by other factors, which may cause the observed band size to be different from the predicted size.

Common causes are:
-Post-translational modification; for example phosphorylation and glycosylation increase the size of the protein
-Post-translation cleavage; many proteins are synthesized as precursor proteins, and then cleaved to give the active form
-Multimers, for example dimerization of a protein. This is usually prevented under reducing conditions, although strong interactions can result in the appearance of higher bands
-Splice variants; alternative splicing may result in different sized proteins being produced from the same gene
-Relative charge; the composition of amino acids (charged vs. non-charged)

Find additional tips, troubleshooting help, and resources within our Protein Electrophoresis and Western Blotting Support Center.

For colorimetric alkaline phosphatase substrates, we offer the following products:

Novex AP Chromogenic Substrate (BCIP/ NBT) (Cat. No. WP20001)

1-Step NBT/BCIP Substrate Solution (Cat. No. 34042)

1-Step PNPP Substrates Solution (Cat. No. 37621)

NBT and BCIP are also available as standalone products:

NBT Substrate Powder (nitro-blue tetrazolium chloride) (Cat. No. 34035)

BCIP Substrate Powder (5-bromo-4-chloro-3'-indolylphosphate p-toluidine) (Cat. No. 34040)

For chemiluminescent alkaline substrates, we offer the following products:

Novex AP Chemiluminescent Substrate (CDP-Star) (Cat. No. WP20002)

NovaBright Phospha-Light EXP Assay Kit for SEAP (Secreted Placental Alkaline Phosphatase) Reporter Gene Detection (Cat. Nos. N10577, N10578)

Find additional tips, troubleshooting help, and resources within our Cell Analysis Support Center.

- Invitrogen AP Chromogenic Substrate (BCIP/NBT) and Invitrogen HRP Chromogenic Substrate (TMB) are ready-to-use, single component substrates for sensitive immunodetection of alkaline phosphatase (AP) or horse radish peroxidase (HRP), respectively, on western blots or dot blots. The sensitivity of the AP substrate is in the 100 pg range, while the HRP substrate is in the 1 ng range (see Note below).

- Invitrogen AP Chromogenic Substrate consists of a ready-to-use solution of 5-bromo-4-chloro-3-indolyl-1-phosphate (BCIP) and nitroblue tetrazolium (NBT) which forms a black-purple precipitate upon reaction with alkaline phosphatase. It is available as a standalone product (Cat. Nos. WP20001) and is also a component of the WesternBreeze Chromogenic Detection Kits.

- Invitrogen HRP Chromogenic Substrate consists of a ready-to-use non-toxic solution of 3,3',5,5'-tetramethylbenzidine (TMB) which forms a blue precipitate upon reaction with horse radish peroxidase. It is available as a standalone product (Cat. No. WP20004).

Note: Sensitivity may vary depending upon the antigen and antibody used for detection.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.

Yes, you can purchase it as a standalone (Cat. No. WP20001). It will give a purple chromogenic precipitate.

Find additional tips, troubleshooting help, and resources within our Protein Assays and Analysis Support Center.