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Vector Laboratories is closed for the President’s Day on Monday, February 19th. We will be back in the office on Tuesday, February 20th.
We will respond to emails upon our return. Have a wonderful day.
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.
| Unit Size | 1 kit |
|---|---|
| Applications | Immunohistochemistry / Immunocytochemistry, In situ hybridization, Blotting Applications, Elispot, ELISAs |
| Detection Enzyme(s) | Peroxidase |
| Substrate Reaction Product | Precipitating Reaction Product |
| Chromogen Color | Blue |
| Microscopy | Brightfield, Spectral Imaging |
| Mounting | Non-Aqueous (Permanent) |
| Safety Title | WARNING: Cancer – www.P65Warnings.ca.gov |
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.
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.
| Substrate | Catalog Number | Hematoxylin (H-3404) Hematoxylin QS (H-3401) | Methyl Green (H-3402) | Nuclear Fast Red (H-3404) |
| ImmPACT DAB (brown) | SK-4105 | Excellent Contrast | Excellent Contrast | Fair Contrast |
| ImmPACT DAB EqV (brown) | SK-4103 | Excellent Contrast | Excellent Contrast | Fair Contrast |
| DAB (brown) | SK-4100 | Excellent Contrast | Excellent Contrast | Fair Contrast |
| DAB-Ni (gray-black) | SK-4100 | Excellent Contrast | Fair Contrast* | Good Contrast |
| ImmPACT AEC (red) | SK-4205 | Excellent Contrast | Counterstain Incompatibility** | Color Incompatibility |
| ImmPACT AMEC Red (red) | SK-4285 | Excellent Contrast | Counterstain Incompatibility** | Color Incompatibility |
| AEC (red) | SK-4200 | Excellent Contrast | Counterstain Incompatibility** | Color Incompatibility |
| TMB (blue) | SK-4400 | Color Incompatibility | Counterstain Incompatibility | Excellent Contrast |
| ImmPACT VIP (purple) | SK-4605 | Fair Contrast | Excellent Contrast | Poor Contrast |
| Vector VIP (purple) | SK-4600 | Fair Contrast | Excellent Contrast | Poor Contrast |
| ImmPACT SG (blue-gray) | SK-4705 | Poor Contrast | Good Contrast | Excellent Contrast |
| SG (blue-gray) | SK-4700 | Poor Contrast | Good Contrast | Excellent Contrast |
| ImmPACT NovaRED (red) | SK-4805 | Excellent Contrast | Excellent Contrast*** | Color Incompatibility |
| Vector NovaRED (red) | SK-4800 | Excellent Contrast | Excellent Contrast*** | Color Incompatibility |
| ImmPACT Vector Red (magenta) | SK-5105 | Excellent Contrast | Excellent Contrast | Color Incompatibility |
| Vector Red (magenta) | SK-5100 | Excellent Contrast | Excellent Contrast | Color Incompatibility |
| Vector Black (black) | SK-5200 | Excellent Contrast | Excellent Contrast* | Excellent Contrast |
| Vector Blue (blue) | SK-5300 | Color Incompatibility | Good Contrast | Excellent Contrast |
| BCIP / NBT (indigo) | SK-5400 | Color Incompatibility | Excellent 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. | ||||
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