Antibody data
- Antibody Data
- Antigen structure
- References [1]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [3]
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Validation data
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- Product number
- PA5-17815 - Provider product page
- 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
- 56 µg/mL
- Storage
- -20°C
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
Frontiers in immunology 2021;12:814570
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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.
- 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
- 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.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Phospho-ZAP70/Syk (Tyr319, Tyr352) using a polyclonal antibody (Product # PA5-17815).
- 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.