Good ABC-AP Staining Kit

February 01, 2017

University of North Carolina at Chapel Hill
Nutrition
Graduate Research Assistant

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Company:

Vector Labs

Product Name:

VECTASTAIN ABC-AP Staining KIT (Alkaline Phosphatase, Guinea Pig IgG)

Catalog Number:

AK-5007

I used this kit for staining after using an anti-cytokeratin 8-18 primary guinea pig Ab (sensitive against mouse) against preneoplastic cells in mouse liver.

Experimental Design and Results Summary

Application

IHC

Starting Material

Paraffin-embedded mouse liver - 10 microns

Protocol Overview

CK 8-18 blocking was performed with goat serum (included in the kit) followed by staining with Vectastain ABC-AP biotinylated anti-guinea pig Ig and ImmPACT Vector Red Alkaline Phosphatase kit. Incubation with Avidin followed by Biotin was performed after goat serum blocking and before primary antibody incubation.

Tips

The kit does not come with biotin blocking, must buy that separately and it is important for liver

Results Summary

There is normal activity characteristic of the primary antibody but the preneoplastic cells with higher than normal activity were visible and stained in a stronger red (see image). Cholangiocytes are positive for cytokeratin 8-18, not to be confounded with preneoplastic hepatocytes.

DOI or PMID #

N/A

Additional Notes

N/A

Image Gallery

Summary

The Good

Easy to use, everything in one kit, blocking buffer included; crisp colors

The Bad

There is nothing bad that can be attributed to this kit as used in this protocol

The Bottom Line

I am familiar with Vector Lab products and I use their secondary staining kits and blocking buffers. The advantage of this kit is that the staining is also fluorescent and it is easy to use.

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