GCET1 Mouse Monoclonal Antibody (RAM341) from Abcam

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Quality of Results

Ease-of-Optimization

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Academic Haematology
Newcastle University
Research Associate
GCET1 (RAM341) IHC staining in diffuse large B-cell lymphoma cells. The antibody demonstrates cytoplasmic staining of large CD20-positive malignant cells.

Company:

Abcam

Product Name:

GCET1 mouse monoclonal antibody (RAM341)

Catalog Number:

ab68889

Image

GCET1 protein has been identified as playing an important role in germinal center B-cell physiology and as such, is deemed a marker for germinal center-like B-cells. I am using the GCET1 antibody to perform immunohistochemical (IHC) staining on diffuse large B-cell lymphoma sections, in order to assess GCET1 protein expression and immunophenotype the tumors as germinal center-like or activated B-cell like according to established IHC algorithms. All IHC is performed using the Ventana® BenchMark Automated Staining System.

Experimental Design and Results Summary

Applications

IHC

Sample

4 micron sections cut from diffuse large B-cell lymphoma tumors

Primary Incubation

Mouse monoclonal (RAM341) antibody to GCET1. The antibody was used at a 1/80 dilution in Ventana Diluent. The protocol comprised a standard CC1 (Ventana) antigen retrieval , 32 minute antibody incubation, Ventana Amplification Kit (Amplifier A&B), 8 minute hematoxylin and 8 minute bluing reagent (Ventana).

Blocking Agent

No blocking agent is required using this antibody in conjunction with the Ventana iVIEW™ DAB Detection Kit.

Secondary Incubation

Biotin conjugated secondary antibody (Ventana iVIEW™ DAB Detection Kit) comprising affinity purified biotinylated goat-anti-mouse IgG and IgM and goat-anti-rabbit IgG.

Tertiary Incubation

N/A

Detection

DAB (3, 3’-diaminobenzidine tetrahydrochloride)

Results Summary

The optimized staining conditions give a moderate strength cytoplasmic staining with minimal background.

Additional Notes

None

Image Gallery

GCET1 (RAM341) IHC staining in diffuse large B-cell lymphoma cells. The antibody demonstrates cytoplasmic staining of large CD20-positive malignant cells.

Summary

The Good

Good value for money with specific staining.

The Bad

Slight background can be a problem and the strength of staining can be weak (amplification required).

The Bottom Line

Good all round antibody with reliable results.

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