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TMB Substrate Kit, Peroxidase (HRP), (3,3', 5,5'-tetramethylbenzidine) (SK-4400)

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Description

TMB (3,3′, 5,5′ tetramethylbenzidine dihydrochloride) is a sensitive chromogen substrate for peroxidase and a non-carcinogenic substitute for benzidine. TMB can be used for blotting, immunohistochemical, and in situ hybridization applications. Slides stained with TMB can be dehydrated and permanently mounted and the soluble blue product, produced in the absence of the stabilizing solution, is useful for ELISA assays.

The TMB HRP Substrate Kit (SK-4400) contains the reagents needed to produce either a precipitating blue reaction product or a soluble blue reaction product. This kit also contains a stabilizing solution which, when added to the working solution, allows the development of a blue precipitate. Kit materials are pre-packaged in convenient dropper bottles.

Features:

  • Greater sensitivity than conventional substrates
  • Stock solutions supplied in convenient dropper bottles promoting ease of handling
  • No wait times for mixing and dissolving powders or tablets
  • Sufficient reagents to produce 300 ml of working solution

Specifications

Unit Size1 kit
ApplicationsImmunohistochemistry / Immunocytochemistry, In situ hybridization, Blotting Applications, Elispot, ELISAs
Detection Enzyme(s)Peroxidase
Substrate Reaction ProductPrecipitating Reaction Product
Chromogen ColorBlue
MicroscopyBrightfield, Spectral Imaging
MountingNon-Aqueous (Permanent)
Safety TitleWARNING: Cancer – www.P65Warnings.ca.gov

What’s in the box:

  • 6 ml TMB Reagent 1
  • 6 ml TMB Reagent 2
  • 6 ml TMB Reagent 3
  • 6 ml TMB Reagent 4

Technical Information

TMB (3,3′, 5,5′ tetramethylbenzidine dihydrochloride) is a sensitive chromogen substrate for peroxidase and a non-carcinogenic substitute for benzidine. TMB can be used for blotting, immunohistochemical, and in situ hybridization applications. Slides stained with TMB can be dehydrated and permanently mounted and the soluble blue product, produced in the absence of the stabilizing solution, is useful for ELISA assays.

 

Counterstain / Substrate Compatibility

This table is designed as a reference to determine the optimal counterstain / substrate combination for your application. Consideration should be given to tissue type, antigen unmasking protocol, and other detection parameters to achieve the desired staining intensity.

SubstrateCatalog NumberHematoxylin (H-3404)
Hematoxylin QS (H-3401)
Methyl Green (H-3402)Nuclear Fast Red (H-3404)
ImmPACT DAB (brown)SK-4105Excellent ContrastExcellent ContrastFair Contrast
ImmPACT DAB EqV (brown)SK-4103Excellent ContrastExcellent ContrastFair Contrast
DAB (brown)SK-4100Excellent ContrastExcellent ContrastFair Contrast
DAB-Ni (gray-black)SK-4100Excellent ContrastFair Contrast*Good Contrast
ImmPACT AEC (red)SK-4205Excellent ContrastCounterstain Incompatibility**Color Incompatibility
ImmPACT AMEC Red (red)SK-4285Excellent ContrastCounterstain Incompatibility**Color Incompatibility
AEC (red)SK-4200Excellent ContrastCounterstain Incompatibility**Color Incompatibility
TMB (blue)SK-4400Color IncompatibilityCounterstain IncompatibilityExcellent Contrast
ImmPACT VIP (purple)SK-4605Fair ContrastExcellent ContrastPoor Contrast
Vector VIP (purple)SK-4600Fair ContrastExcellent ContrastPoor Contrast
ImmPACT SG (blue-gray)SK-4705Poor ContrastGood ContrastExcellent Contrast
SG (blue-gray)SK-4700Poor ContrastGood ContrastExcellent Contrast
ImmPACT NovaRED (red)SK-4805Excellent ContrastExcellent Contrast***Color Incompatibility
Vector NovaRED (red)SK-4800Excellent ContrastExcellent Contrast***Color Incompatibility
ImmPACT Vector Red (magenta)SK-5105Excellent ContrastExcellent ContrastColor Incompatibility
Vector Red (magenta)SK-5100Excellent ContrastExcellent ContrastColor Incompatibility
Vector Black (black)SK-5200Excellent ContrastExcellent Contrast*Excellent Contrast
Vector Blue (blue)SK-5300Color IncompatibilityGood ContrastExcellent Contrast
BCIP / NBT (indigo)SK-5400Color IncompatibilityExcellent Contrast*Excellent Contrast
*This substrate shows a slight decrease in sensitivity following the methyl green protocol. This decrease can be minimized by reducing the heat incubation and acetone rinse times.
**Substrate dissolves in acetone wash.
***A slight color change in ImmPACT NovaRED and Vector NovaRED reaction produt may be seen using methyl green.

Substratesensitivity Jpg Substratesensitivity

Applicable patents and legal notices are available at legal notices.

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