P21 Mouse Monoclonal Antibody Clone 4D10 from Abcam

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Academic Haematology
Newcastle University
Research Associate
p21 (4D10) IHC staining in diffuse large B-cell lymphoma cells. The antibody demonstrates nuclear staining of large CD20-positive malignant cells.

Company:

Abcam

Product Name:

P21 Mouse Monoclonal Antibody Clone 4D10

Catalog Number:

ab49407

Image

The p21 protein is regulated by MYC and functions as a regulator of cell cycle progression. p21 expression is also controlled by the tumor suppressor protein p53, through which it mediates p53-dependent cell cycle arrest. P21 has been implicated in the pathogenesis of diffuse large B-cell lymphoma (DLBCL). I am using the p21 antibody to measure p21 protein levels by immunohistochemistry (IHC) in DLBCL formalin fixed paraffin embedded tissue (FFPET) sections.

Experimental Design and Results Summary

Applications

IHC

Sample

4 micron sections cut from DLBCL tumors

Primary Incubation

Mouse monoclonal (4D10) antibody to p21 (cat# ab49407). The antibody was used at a 1/5 dilution in Ventana® Diluent (cat #760-108). The protocol comprised Ventana cell conditioning 1 (CC1) (cat #950-124) based pre-treatment step for antigen retrieval, 40 minute antibody incubation, 8 minute hematoxylin (Ventana, cat #760-2021) and 8 minute bluing reagent (Ventana, cat #760-2037) in conjunction with the Ventana ultraView™ DAB Detection Kit (cat #760-500).

Blocking Agent

No blocking agent is required using this antibody with the Ventana ultraView DAB Detection Kit.

Secondary Incubation

Universal HRP Multimer-containing HRP-labeled antibodies; goat anti-mouse IgG, goat anti-mouse IgM and goat anti-rabbit IgG (antibodies are included in the Ventana ultraView DAB Detection Kit). These antibodies are supplied in a ready-to-use pre-diluted automatic dispenser and the cocktail is applied to each slide.

Tertiary Incubation

N/A

Detection

DAB (3, 3’-diaminobenzidine tetrahydrochloride)

Results Summary

The optimized staining conditions give specific nuclear staining.

Additional Notes

None.

Image Gallery

p21 (4D10) IHC staining in diffuse large B-cell lymphoma cells. The antibody demonstrates nuclear staining of large CD20-positive malignant cells.

Summary

The Good

Nuclear staining is specific and clear.

The Bad

Requires a high concentration and thus, is relatively expensive.

The Bottom Line

Excellent all round antibody with specific results, but costly.

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