Assay & Detection

(AP) Anti-Mouse (Permanent Red) Kit

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(AP) Anti-Mouse (Permanent Red) Kit

Article No

KDB-10020

Application

IHC

Size

1 Kit (8ml / 80 Tests)

Product insert

Suggested protocols

Specifications

Application IHC
Article No KDB-10020
BioSite Brand BioSite Histo
Conjugation AP
Description (AP) Anti-Mouse (Permanent Red) Kit
Supplier BioSite Histo
Notes High sensitivity streptavidin biotin detection kits for mouse primary antibodies. 8 ml Kits including chromogen. Ready-to-use blocking solution, sec. antibody, enzym conjugate.
Product Type Assay & Detection
Protocol Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary or secondary antibody is minimized by incubation with a protein blocking solution (“Blocking Solution” provided with the kit). This step can be omitted if the primary antibodies are diluted in an appropriate buffer. The next step is incubation with the specific primary antibody. After washing, the biotinylated secondary antibody is applied and incubated. This secondary antibody functions as a link between primary antibody and the streptavidinalkaline phosphatase-conjugate (“Streptavidin-AP-Conjugate“). A second washing is followed by the application of this conjugate. It binds to the biotin at the secondary antibody. Any excess of unbound streptavidin-AP-conjugate is thoroughly washed away after incubation. The addition of the chromogenic substrate starts the enzymatic reaction of the alkaline phosphatase which leads to colour precipitation where the primary antibody is bound. The colour can be observed with a light microscope. The chromogen used determines the colour. The chromogen Fast Red (included only in kit KDB-10020) leads to a magenta-red product of reaction at the place of the target antigen. Other suitable chromogens are Permanent AP Red (magenta-red), New Fuchsin (magenta-red) or NBT (blue-black) with its substrate BCIP. Staining procedure 1. Blocking Solution (protein block, Reagent 1) (This step is optional.) 5 min. 2. Washing with wash buffer 1 x 2 min. 3. Primary antibody (optimally diluted) or negative control reagent 30-60 min. 4. Washing with wash buffer 3 x 2 min. 5. Biotinylated Secondary Antibody, Mouse (Reagent 2, yellow) 10-15 min. 6. Washing with wash buffer 3 x 2 min. 7. Streptavidin-AP-Conjugate (Reagent 3, red) 10-15 min. 8. Washing with wash buffer 3 x 2 min. 9. Fast Red, Permanent AP Red, NBT/BCIP or New Fuchsin 10-20 min. (Controlling the colour intensity via light microscope is recommended.) 10. Stopping the reaction with distilled H2O when the desired colour intensity is attained 11. Counterstaining and blueing 12. Mounting: aqueous with Fast Red, permanent with Permanent AP Red, NBT/BCIP or New Fuchsin
References Elias JM “Immunohistopathology – A practical Approach to Diagnosis” ASCP Press 2003 Nadji M and Morales AR Ann N.Y. Acad Sci 420:134-9, 1983
Size 1 Kit (8ml / 80 Tests)
Storage 2 - 8°C
Technical Specifications High sensitivity broad spectrum streptavidin biotin detection kits for mouse primary antibodies. 8 ml kits including chromogen. Ready-to-use blocking solution, sec. antibody, enzyme conjugate.
Product Page Updated 2019-08-15T09:29:01.207Z

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