Novex™ AP Chromogenic Substrate (BCIP/ NBT)

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.