MA1-10182

antibody from Invitrogen Antibodies

Targeting: CD247 CD3H, CD3Q, CD3Z

Western blot (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Recommended by provider)

Flow cytometry (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-10182

Provider

Invitrogen Antibodies

Product name

CD3e Monoclonal Antibody (APA1/1)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

This antibody recognizes an activation-dependent intracellular epitope of CD3 epsilon. Exposure of the epitope precedes CD3 phosphorylation and recruitment and activation of ZAP70, which initiates the signaling cascade produced by T-cell activation. APA1/1 provides the earliest known marker for TCR-mediated T cell activation. Immunocytochemistry: Fixed and permeabilized cells. The antibody can distinguish TCR-stimulated from non-stimulated cells.

Reactivity

Human, Mouse

Host

Mouse

Isotype

IgG

Antibody clone number

APA1/1

Vial size

100 µg

Concentration

1 mg/mL

Storage

4° C, do not freeze

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-CD3z Monoclonal Antibody (APA1/1) (Product # MA1-10182) and an ~18 kDa band corresponding to T-cell surface glycoprotein CD3 zeta chain, along with an uncharacterized band (*) was observed across positive cell line Jurkat; and not across negative cell lines HeLa,SK-OV-3 and Raji. Membrane enriched extracts (30 µg lysate) of Jurkat (Lane 1), HeLa (Lane 2), SK-OV-3 (Lane 3) and Raji (Lane 4) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0341BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Separation of Jurkat cells stained anti-human CD3 activation epitope (APA1/1) purified Monoclonal antibody (Product # MA1-10182) (concentration in sample 5 µg/mL, GAM FITC, red-filled) from Jurkat cells unstained by primary antibody (GAM FITC, black-dashed) in flow cytometry analysis (intracellular staining).