TMB 1-Component HRP Membrane Substrate (SUBM)

About this product

A ready-to-use formulation containing TMB in a mildly acidic buffer. This TMB substrate is useful for membrane-based immunoblotting applications using HRP-conjugates for detection. Yields an insoluble, permanent dark blue reaction product.

Specifications

Catalog number

6280

Size / count

100 mL

Dry ice

No

Hazardous

No

Discontinued

Yes

Label

ICT

Type

ELISA & Assay Reagents

Assay Protocol

1. Allow product to equilibrate to room temperature (25°C) prior to use., 2. Completely cover the membrane surface with the substrate. The substrate will react with phosphatase creating a purple-blue reaction product., 3. To stop the reaction, rinse the developed membrane with distilled water., 4. For best results, the reaction should be monitored and read before the background staining becomes too intense to distinguish from the positive staining.

Concentration

1X

pH

pH: 5.25-5.65

Additionnal Information

ELISA & Assay Reagents ; Substrates

Shipping Conditions

Ships overnight (domestic), International Priority Shipping

Storage Temperature

2-8°C

Notes

20% discount

Shelf Life

3 years from date of manufacture

Target Description

TMB 1-Component HRP Membrane Substrate (SUBM) is suitable for membrane applications using horseradish peroxidase (HRP) as the conjugated detection enzyme. SUBM should not be used for microWell (ELISA) applications. TMB 1 Component HRP Membrane Substrate is a one-component formulation containing stabilized 3,3’,5,5’-tetramethylbenzidine (TMB). The TMB substrate is oxidized by the peroxidase enzyme to yield an insoluble dark blue reaction product. SUBM is supplied ready-to-use at 1X. Best results are obtained by equilibrating TMB 1-Component HRP Membrane Substrate to room temperature (25°C) prior to use. After probing with the antibody and HRP reagents, wash membrane thoroughly and transfer the membrane into a clean container. Cover the membrane surface with ample amount of SUBM and incubate 5-20 minutes. The substrate will react with sites on the membrane containing peroxidase, producing an insoluble permanent dark blue reaction product. For best results, monitor the substrate color development process until the target protein bands are visible. To stop the reaction, rinse the membrane with reagent quality water. If the reaction proceeds too long, there will be excessive background staining and diminished resolution of the target peptide or protein banding regions. If the color development is too rapid or intense, it is recommended to dilute the antibodies or conjugates or shorten the incubation period.