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Western blot
ELISAAntibody data
- Antibody Data
- Antigen structure
- References [3]
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- Validations
- Western blot [2]
- Immunocytochemistry [1]
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- Product number
- 13-0600 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Zap-70 Monoclonal Antibody (ZAP-70-6F7)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human, Mouse
- Host
- Mouse
- Isotype
- IgG
- Antibody clone number
- ZAP-70-6F7
- Vial size
- 100 µg
- Concentration
- 0.5 mg/mL
- Storage
- -20°C
Submitted references Lck dephosphorylation at Tyr-394 and inhibition of T cell antigen receptor signaling by Yersinia phosphatase YopH.
Activation of ZAP-70 through specific dephosphorylation at the inhibitory Tyr-292 by the low molecular weight phosphotyrosine phosphatase (LMPTP).
Negative regulation of T-cell activation and autoimmunity by Mgat5 N-glycosylation.
Alonso A, Bottini N, Bruckner S, Rahmouni S, Williams S, Schoenberger SP, Mustelin T
The Journal of biological chemistry 2004 Feb 6;279(6):4922-8
The Journal of biological chemistry 2004 Feb 6;279(6):4922-8
Activation of ZAP-70 through specific dephosphorylation at the inhibitory Tyr-292 by the low molecular weight phosphotyrosine phosphatase (LMPTP).
Bottini N, Stefanini L, Williams S, Alonso A, Jascur T, Abraham RT, Couture C, Mustelin T
The Journal of biological chemistry 2002 Jul 5;277(27):24220-4
The Journal of biological chemistry 2002 Jul 5;277(27):24220-4
Negative regulation of T-cell activation and autoimmunity by Mgat5 N-glycosylation.
Demetriou M, Granovsky M, Quaggin S, Dennis JW
Nature 2001 Feb 8;409(6821):733-9
Nature 2001 Feb 8;409(6821):733-9
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Raji (lane 1) and Jurkat (lane 2) cells using Ms anti-ZAP-70 (Product # 13-0600)
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of Jurkat (Lane 1) and MOLT-4 (Lane 2). The blot was probed with Anti-ZAP70 Mouse Monoclonal Antibody (Product # 13-0600, 2 µg/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 µg/mL, 1:2500 dilution). A 70 kDa band corresponding to ZAP70 was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody following blocking with 5 % skimmed milk. Chemiluminescent detection was performed using Pierce™ ECL Western Blotting Substrate (Product # 32106).
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescence analysis of ZAP70 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 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with ZAP70 (ZAP-70-6F7) Mouse Monoclonal Antibody (Product # 13-0600) at 2µg/mL in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A28175) 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 Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
