505-0049-42

antibody from Invitrogen Antibodies

Targeting: CD4

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

505-0049-42

Provider

Invitrogen Antibodies

Product name

CD4 Monoclonal Antibody (RPA-T4), Alexa Fluor™ 561, eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The RPA-T4 monoclonal antibody reacts with human CD4, a 59 kDa cell surface receptor expressed by a majority of thymocytes, subpopulation of mature T cells (T-helper cells) and in low levels on monocytes. CD4 is a receptor for the human immunodeficiency virus (HIV). RPA-T4 blocks HIV binding and mixed lymphocyte reaction. The RPA-T4 antibody recognizes a different epitope than the OKT4 monoclonal antibody, and these antibodies do not cross-block binding to each other's respective epitopes. Applications Reported: This RPA-T4 antibody has been reported for use in flow cytometric analysis. Applications Tested: This RPA-T4 antibody has been pre-diluted and tested by flow cytometric analysis of normal human peripheral blood cells. This may be used at 5 µL (0.25 µg) per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. Alexa Fluor 561 emits at 575 nm and is intended for use on spectral cytometers where it may be multiplexed with both PE and PE-eFluor 610. Excitation: 558 nm; Emission: 575 nm; Laser: Yellow-Green Laser

Reactivity

Human

Host

Mouse

Isotype

IgG

Antibody clone number

RPA-T4

Vial size

100 Tests

Concentration

5 µL/Test

Storage

4° C, store in dark, DO NOT FREEZE!

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Normal human peripheral blood cells were stained with CD3 Monoclonal Antibody, Alexa Fluor 532 (Product # 58-0038-42), CD19 Monoclonal Antibody, FITC (Product # 11-0199-42 ), CD8a Monoclonal Antibody, PE (Product # 12-0088-42), CD25 Monoclonal Antibody, PE-Cyanine7 (Product # 25-0259-42), CD56 (NCAM) Monoclonal Antibody, PE-eFluor 610 (Product # 61-0567-42) and Mouse IgG1 kappa Isotype Control, Alexa Fluor 561 (Product # 606-4714-81) or CD4 Monoclonal Antibody, Alexa Fluor 561. Live cells as determined by Fixable Viability Dye eFluor 450 (Product # 65-0863-14) in the lymphocyte gate were used for analysis and first gated on CD3 positive/CD19 negative cells (top left). CD3 positive cells were analyzed for their expression of CD4 and CD8a (top right), CD4 and CD56 (NCAM) (bottom left) and CD4 and CD25 (bottom right). This data was collected on a 5-laser Cytek Aurora full spectral cytometer. As shown here, Alexa Fluor 561 is compatible with PE and PE tandems when proper panel design principles are applied.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Normal human peripheral blood cells were stained with CD3 Monoclonal Antibody, FITC (Product # 11-0037-42) and Mouse IgG1 kappa Isotype Control, Alexa Fluor 561 (Product # 505-4714-81) (left) or CD4 Monoclonal Antibody, Alexa Fluor 561 (right). Viable cells in the lymphocyte gate were used for analysis, as determined by 7-AAD (Product # 00-6993-50). This data was collected on a 5-laser Cytek Aurora full spectral cytometer.