Assay & Detection

One-Step Polymer anti-Mouse/Rabbit/Rat

NEW
(AP) One-Step Polymer anti-Mouse/Rabbit/Rat

Article No

KDB-10047

Application

IHC

Species Reactivity

mouse
rabbit
rat

Size

100 ml / 1000 Tests

Product insert

Suggested protocols

Specifications

Application IHC
Article No KDB-10047
BioSite Brand BioSite Histo
Conjugation AP
Description (AP) One-Step Polymer anti-Mouse/Rabbit/Rat
Supplier BioSite Histo
Notes Enzym polymer single reagents with alkaline phosphatase. Ready-to-use
Product Type Assay & Detection
Protocol Paraffin-embedded tissue sections are first deparaffinised and rehydrated. Background staining caused by unspecific binding of the primary antibody or the secondary antibody in the AP One-Step Polymer is minimized by incubation with a protein blocking solution. 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 AP One-Step Polymer is applied and incubated. Any excess of unbound polymer 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 Permanent AP Red leads to the formation of a magentared product of reaction at the place of the target antigen. Other suitable chromogens are Fast Red (magenta-red), New Fuchsin (magenta-red), or NBT (blue-black) with its substrate BCIP. Staining procedure 1. Blocking Solution (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 5 min. 5. AP One-Step Polymer anti-Mouse/Rabbit/Rat 30 Min. 6. Washing with wash buffer 3 x 2 min. 7. Permanent AP Red, Fast Red, NBT/BCIP or New Fuchsin 10 - 20 min. (Controlling the colour intensity via light microscope is recommended.) 8. Stopping the reaction with distilled H2O when the desired colour intensity is attained 9. Counterstaining and blueing 10. 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 100 ml / 1000 Tests
Species Reactivity mouse, rabbit, rat
Storage 2 - 8°C
Technical Specifications Single step polymer detection kit with alkaline phoshatase
Product Page Updated 2019-08-15T09:29:01.207Z

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