MA5-15180

antibody from Invitrogen Antibodies

Targeting: ZAP70 SRK, STD, ZAP-70

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Immunoprecipitation (Recommended by provider)

Immunohistochemistry (Supportive data in Antibodypedia)

Flow cytometry (Recommended by provider)

Antibody Data

Product number

MA5-15180

Provider

Invitrogen Antibodies

Product name

Zap-70 Monoclonal Antibody (J.947.4)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

It is not recommended to aliquot this antibody.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Antibody clone number

J.947.4

Vial size

100 µL

Concentration

142 µg/mL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of Zap-70 in extracts from Jurkat cells, starved for 16 hours and either untreated or H2O2-treated (2 mM for 3 minutes), using Zap-70 monoclonal antibody (Product # MA5-15180).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-Zap-70 Monoclonal Antibody (J.947.4)(Product # MA5-15180) and a 70kDa band corresponding to Zap-70 was observed across cell lines and tissues tested. Membrane enriched extracts (30 µg lysate) of Jurkat (Lane 1), MOLT-4 (Lane 2), Raji (Lane 3), Ramos (Lane 4), EL4 (Lane 5) and MEF (Lane 6). Tissue Extracts of Mouse Thymus (Lane 7), Rat Thymus (Lane 8), Mouse Liver (Lane 9) and Rat Liver (Lane 10) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0322BOX). 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-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Zap-70 was performed using 70 confluent log phase Jurkat cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with Zap-70 Monoclonal Antibody (J.947.4) (Product # MA5-15180) at 1:100dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b: Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300 dilution). Panel d represents the merged image showing membrane localization. Panel e represents Raji cells with no expression of Zap-70. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

Immunohistochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunohistochemical analysis of ZAP70 using a monoclonal antibody (Product # MA5-15180).