Antibody data
- Antibody Data
- Antigen structure
- References [0]
- Comments [0]
- Validations
- Western blot [1]
- Immunohistochemistry [1]
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Validation data
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- Product number
- 44-230G - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-Zap-70 (Tyr292) Polyclonal Antibody
- Antibody type
- Polyclonal
- Antigen
- Synthetic peptide
- Reactivity
- Human
- Host
- Rabbit
- Isotype
- IgG
- Vial size
- 100 µL
- Storage
- -20°C
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Lysates prepared from Jurkat cells left untreated (1) or treated with hydrogen peroxide (2-6) were resolved by SDS-PAGE on a 10% polyacrylamide gel and transferred to PVDF. Membranes were either left untreated (1-5) or treated with lambda (λ) phosphatase (6), blocked with a 5% BS A-TBST buffer for one hour at room temperature, and incubated with ZAP-70 (pY292) antibody for two hours at room temperature in a 3% BSA-TBST buffer, following prior incubation with: no peptide (1, 6), the non-phosphopeptide corresponding to the immunogen (2), a generic phosphotyrosine-containing peptide (4) or, the phosphopeptide immunogen (5). After washing, membranes were incubated with goat F (ab')2 anti-rabbit IgG HRP conjugate (Product # ALI4404), and bands were detected using the Pierce SuperSignal™ method.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of Phospho-ZAP-70 pTyr292 showing staining in the cytoplasm of paraffin-embedded human lymph node tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Anti- Phospho-ZAP-70 pTyr292 Polyclonal Antibody (Product # 44-230G) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.