Assay & Detection

(AP) Polymer anti-Mouse

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(AP) Polymer anti-Mouse

Article No

KDB-30303

Application

IHC

Species Reactivity

mouse

Size

100 ml / 1000 Tests

Suggested protocols

Specifications

Application IHC
Article No KDB-30303
BioSite Brand BioSite Histo
Conjugation AP
Description (AP) Polymer anti-Mouse
Supplier BioSite Histo
Format Ready-To-Use
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 polymer is minimized by incubation with a protein blocking solution (“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-polymer is applied and incubated. Any excess of unbound AP-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 Fast Red leads to the formation of 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. 1. Blocking Solution (protein block, 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-polymer anti Mouse 30 min. 6. Washing with wash buffer 3 x 2 min. 7. Fast Red, Permanent AP Red, NBT/BCIP or New Fuchsin 5-15 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
Storage The solution should be stored at 2-8°C without further dilution. Please store the reagent in a dark place and do not freeze it. Under these conditions the solution is stable up to the expiry date indicated on the label. It should not be used after the expiry date.
Technical Specifications Enzyme single polymer reagents with alkaline phoshatase. Ready-to-use.
Product Page Updated 2019-04-10T07:46:12.355Z

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