TMB HRP Membrane Substrate One Component ‘Ready To Use’

Pricing & Details

Product No.T116
Formats AvailableView All
Product Type
Substrate and Assay Reagents
Applications
WB
Prod No.
Size
Price
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T116-100 ml
100 ml
$50.00
In stock
Max:
Min: 1
Step: 1
T116-1.0 L
1.0 L
$350.00
In stock
Max:
Min: 1
Step: 1

Product Details

Storage and Handling
Store at temperatures from 2-25ºC. Do Not Freeze. The substrate should be protected from direct light by storing in amber bottles. Only high quality amber glass and plastic products should be used for storing aliquots.
Expiration Date
Stable for a minimum of 30 months from the manufactured date when properly stored at 2-25ºC.

Description

Background
TMB HRPO Membrane Substrate (3,3’,5,5’ tetramethylbenzidine) is a precipitating substrate used with the enzyme horseradish peroxidase (HRPO) designed for various membrane immunoassays but not recommended for microwell or immunohistochemical applications. Initially, the substrate should be slightly yellow to pink in color and is formulated in a mildly acidic buffer. TMB HRPO Membrane Substrate turns a deep blue color when reacted with horseradish peroxidase labeled conjugates.

Leinco Technologies’ TMB HRPO Membrane Substrate is a highly sensitive, high performance formulation that produces a high signal to noise ratio with low membrane background. The outstanding shelf life of at least thirty months for the TMB Membrane Substrate makes this reagent ideal for long term use of the same manufacturing lot.
Directions for Use
TMB HRPO Membrane Substrate is a ready to use solution that needs no preparation or dilution. It is recommended that you allow the substrate solution to equilibrate to room temperature before use. Pour the necessary amount of substrate onto the blocked HRPO conjugate labeled membrane to completely cover the membrane. The substrate will react with sites on the membrane containing peroxidase, producing an insoluble permanent dark blue reaction product. The reaction should be monitored and read before the background color becomes too intense which may result in diminished contrast between the positive and background staining. To stop the reaction, rinse the membrane with reagent quality water. If a less intense reaction is desired, reduce the concentration of the HRPO labeled conjugate. Do not dilute the substrate.
Troubleshooting
 
Problem Cause Solution
Too much background signal observed · BCIP/NBT substrate was left on the membrane too long · Decrease the amount of time the BCIP/NBT substrate is on the membrane
· Too much primary antibody used · Decrease the amount of primary antibody used and wash the membrane sufficiently after the primary antibody incubation
· Too much secondary antibody used · Decrease the amount of secondary antibody used
Nonspecific bands show up on the membrane · Too much primary antibody used · Decrease the amount of primary antibody used and wash the membrane sufficiently after the primary antibody incubation
· Too much secondary antibody used · Decrease the amount of secondary antibody used
Signal disappears from membrane · Membrane not stored correctly · Store the membrane dry in plastic bag in the dark
No signal is observed on the membrane · Low amounts of specific signal present · Expose the membrane to BCIP/NBT substrate for a longer period of time. Include positive control(s) during analysis
· Insufficient primary antibody used · Use more primary antibody
· Insufficient secondary antibody used · Use more secondary antibody
· Sample degraded · Add protease inhibitors to original sample before running a gel

Related Protocols

General Western Blot Protocol
Products are for research use only. Not for use in diagnostic or therapeutic procedures.