MA5-14432

antibody from Invitrogen Antibodies

Targeting: CD8B CD8B1

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-14432

Provider

Invitrogen Antibodies

Product name

CD4/CD8 Antibody Cocktail

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

This CD4/CD8 monoclonal cocktail contains Mouse HRP CD4 (Clone 4B12, IgG1 kappa) and Rabbit AP CD8 (Clone SP16, IgG). It is designed for qualitative immunohistochemistry with normal and neoplastic formalin-fixed, paraffin-embedded tissues using the MultiVision anti-mouse/HRP + anti-rabbit/AP detection system (Catalog # TL-012-MHRA). This cocktail is provided at an optimal concentration in an easy-to-use format for visualizing CD4 and CD8 lymphocyte subsets in red and blue, respectively. The use of rabbit monoclonal CD8 allows for superior staining, in both sensitivity and specificity, permiting the use of higher titers compared to mouse monoclonal antibodies. These cocktails are specially formulated from MultiVision polymers that provide increased sensitivity, time-savings, and detection simplicity. The MultiVision AP and HRP polymers are innovative, patented technology. The smaller amino acid polymer subunits minimize conflicts in binding the target protein resulting in more consistent staining and better signal amplification. Ultimately, this gives the user higher sensitivity and antibody efficiency as well as better signal-to-noise ratios. This system is biotin-free, which prevents background staining found with traditional biotin-based methods. The MultiVision polymer detection systems yield high quality double staining in less than 2 hours on formalin-fixed and paraffin-embedded tissue sections. Prior to staining, heat-induced epitope recovery (HIER) is recommended, using 10mM EDTA (pH 8.0) for 20min at 98ºC followed by cooling at room temperature for 20min. The kits allow observation of two single-stained cells in red and blue and/or co-localization marked by a purple/brownish mixed-color.

Reactivity

Human

Isotype

IgG

Antibody clone number

4B12, SP16

Vial size

7 mL

Storage

4° C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of CD4/CD8 was performed by loading 25 ug of CEM (lane 1), Jurkat (lane 2) and CTLL-2 (lane 3) cell lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4ºC overnight. The membrane was probed with a CD4/CD8 monoclonal antibody (Product # MA5-14432) at a dilution of 1:2 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~55 and 32 kDa.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Formalin-fixed paraffin-embedded human tonsil stained with the MultiVision anti-Mouse/ HRP + anti-Rabbit/ AP. Note: Cell membrane staining of CD4 (in red) and CD8 (in blue).