PA5-17778

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)

Antibody Data

Product number

PA5-17778

Provider

Invitrogen Antibodies

Product name

Phospho-Zap-70 (Tyr493) Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

It is not recommended to aliquot this antibody. This antibody also cross-reacts with endogenous levels of Syk when phosphorylated at Tyr526.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

55 µg/mL

Storage

-20°C

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of MOLT-4 (Lane 1) and MOLT-4 treated with Pervanadate (100 µM for 10 minutes) (Lane 2). The blot was probed with Anti-Phospho-Zap-70 (Tyr493) Polyclonal Antibody (Product # PA5-17778, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 70 kDa band corresponding to Phospho-Zap-70 (Tyr493) was observed to be induced upon Pervanadate treatment in MOLT-4 cells.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of MOLT-4 (Lane 1) and MOLT-4 treated with Pervanadate (100 µM for 10 minutes) (Lane 2). The blot was probed with Anti-Phospho-Zap-70 (Tyr493) Polyclonal Antibody (Product # PA5-17778, 1:1000 dilution) and detected by chemiluminescence using Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A27036, 0.25 µg/mL, 1:4000 dilution). A 70 kDa band corresponding to Phospho-Zap-70 (Tyr493) was observed to be induced upon Pervanadate treatment in MOLT-4 cells.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Phospho-ZAP70 (Tyr493) was performed using 70% confluent log phase MOLT-4 cells untreated or treated with 100 µM Pervanadate for 10 minutes. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with anti-Phospho-ZAP70 (Tyr493) Rabbit Polyclonal Antibody (Product # PA5-17778) at 1:200 dilution in 0.1% BSA, incubated at 4 degree Celsius overnight and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) at a dilution of 1:2000 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). Panel d represents the merged image. Panel e represents untreated control cells while Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification