p504s Antibody – Mouse Monoclonal

Monoclonal

IgG

IHC053

Mouse

Human

Liquid

Store at 2-8 degree C. Do not freeze.

Tris Buffer, ph7.4, 1% BSA, <0.1% NaN3

Colon Carcinoma

1. Tissues are fixed with 10% neutral buffered formalin, cut to ~4um, and placed on a positively charged glass slide.
2. The prepared slide is baked for at least 30 minutes and at most 24 hours at 55-65 degree C.
3. Pretreatment - Perform heat-induced epitope retrieval (HIER) at pH 6 for 10 to 30 minutes.
4. Block – Block with peroxidase solution for 5 minutes at RT.
5. Primary Ab – Dilute antibodies 1:100 – 1:200 and incubate for 10 - 30 minutes at RT.
6. Secondary Ab - Incubate for 20 - 30 minutes at RT.
7. Substrate - Incubate DAB or Fast Red for 5 - 10 minutes at RT.
8. Counterstain - Counterstain with hematoxylin for 30 seconds to 5 minutes, rinse with distilled water and blueing solution for 30 seconds.
9. Dehydrate and apply coverslip.
(Each researcher should optimize the protocol for their experimental model system.)