PA5-17815

antibody from Invitrogen Antibodies

Targeting: ZAP70 SRK, STD, ZAP-70

Western blot (Supportive data in Antibodypedia)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Recommended by provider)

Antibody Data

Product number

PA5-17815

Provider

Invitrogen Antibodies

Product name

Phospho-ZAP70/Syk (Tyr319, Tyr352) Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Synthetic peptide

Description

It is not recommended to aliquot this antibody.

Reactivity

Human

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

37 µg/mL

Storage

-20°C

References

Submitted references

Sorting Nexin 27 Enables MTOC and Secretory Machinery Translocation to the Immune Synapse.

González-Mancha N, Rodríguez-Rodríguez C, Alcover A, Merida I
Frontiers in immunology 2021;12:814570

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 Ramos (Lane 1), Ramos treated with Hydrogen Peroxide (1.5mM for 5 mins) followed by Sodium Orthovanadate treatment (Lane 2), Jurkat (Lane 3), Jurkat treated with Hydrogen Peroxide (1.5mM for 5 mins) followed by Sodium Orthovanadate treatment (Lane 4), THP-1 (Lane 5), THP-1 treated with Hydrogen Peroxide (1.5mM for 5 mins) followed by Sodium Orthovanadate treatment (Lane 6), Raji (Lane 7) and Raji treated with Hydrogen Peroxide (1.5mM for 5 mins) followed by Sodium Orthovanadate treatment (Lane 8). The blot was probed with Anti-Phospho-ZAP70 (Tyr319, Tyr352) Polyclonal Antibody (Product # PA5-17815, 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-ZAP70 (Tyr319, Tyr352) was observed only in the treated cell lines tested.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on whole cell extracts (30 µg lysate) of Ramos (Lane 1), Ramos treated with Hydrogen Peroxide (1.5mM for 5 mins) followed by Sodium Orthovanadate treatment (Lane 2), Jurkat (Lane 3), Jurkat treated with Hydrogen Peroxide (1.5mM for 5 mins) followed by Sodium Orthovanadate treatment (Lane 4), THP-1 (Lane 5), THP-1 treated with Hydrogen Peroxide (1.5mM for 5 mins) followed by Sodium Orthovanadate treatment (Lane 6), Raji (Lane 7) and Raji treated with Hydrogen Peroxide (1.5mM for 5 mins) followed by Sodium Orthovanadate treatment (Lane 8). The blot was probed with Anti-Phospho-ZAP70 (Tyr319, Tyr352) Polyclonal Antibody (Product # PA5-17815, 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-ZAP70 (Tyr319, Tyr352) was observed only in the treated cell lines tested.

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of Phospho-Zap-70 pTyr319/Syk pTyr352 in Jurkat cells, CD3-treated, using a Phospho-Zap-70 pTyr319/Syk pTyr352 polyclonal antibody (Product # PA5-17815) (red). DNA is labeled using a fluorescent blue dye.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescent analysis of Phospho-ZAP70/Syk (Tyr319, Tyr352) using a polyclonal antibody (Product # PA5-17815).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Phospho-ZAP70/Syk (Tyr319, Tyr352) was performed using 70% confluent log phase Ramos cells treated with 1.5mM of Hydrogen peroxide+ 0.5mM of Sodium ortho vanadate for 5 mins. 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 Phospho-ZAP70/Syk (Tyr319, Tyr352) Rabbit Polyclonal Antibody (Product # PA5-17815) at 1:100 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 SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows untreated cells with reduced signal. Panel f represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.