ZAP70 Recombinant Rabbit Monoclonal Antibody [PSH11-52]
cat.: HA723344
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: PSH11-52
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 70 kDa
Isotype: IgG
Immunogen: Synthetic peptide within human ZAP70 aa 301-350.
Positive control: Jurkat cell lysate, MOLT-4 cell lysate, HUT 102 cell lysate, Jurkat, human lymph node tissue, human spleen tissue, mouse lymph node tissue, mouse spleen tissue, rat lymph node tissue, rat spleen tissue.
Subcellular location: Cytoplasm. Cell membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:2,000
1:500
1:1,000
1:1,000
Uniprot #: SwissProt: P43403 Human | P43404 Mouse
Entrez Gene: 301348 Rat
Alternative names: 70 kDa zeta associated protein 70 kDa zeta-associated protein EC 2.7.10.2 FLJ17670 FLJ17679 Selective T cell defect SRK STD Syk related tyrosine kinase Syk-related tyrosine kinase Truncated ZAP kinase Tyrosine protein kinase ZAP70 Tyrosine-protein kinase ZAP-70 TZK ZAP 70 ZAP70 ZAP70_HUMAN Zeta chain associated protein kinase 70kD Zeta chain associated protein kinase 70kDa Zeta chain associated protein kinase 70kDa isoform 1 Zeta chain associated protein kinase 70kDa isoform 2 Zeta chain of T cell receptor associated protein kinase 70 Zeta chain TCR associated protein kinase 70kD Zeta chain TCR associated protein kinase 70kDa
Images
HA723344_1.jpg Fig1: Western blot analysis of ZAP70 on different lysates with Rabbit anti-ZAP70 antibody (HA723344) at 1/2,000 dilution.

Lane 1: Jurkat cell lysate
Lane 2: Ramos cell lysate (negative)
Lane 3: MOLT-4 cell lysate
Lane 4: Raji cell lysate (negative)
Lane 5: HUT 102 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 70 kDa
Observed band size: 70 kDa

Exposure time: 6 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA723344) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA723344_2.jpg Fig2: Immunocytochemistry analysis of Jurkat (positive) and Ramos (negative) labeling ZAP70 with Rabbit anti-ZAP70 antibody (HA723344) at 1/500 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-ZAP70 antibody (HA723344) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA723344_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human lymph node tissue with Rabbit anti-ZAP70 antibody (HA723344) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723344) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723344_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-ZAP70 antibody (HA723344) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723344) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723344_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human liver tissue (negative) with Rabbit anti-ZAP70 antibody (HA723344) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723344) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723344_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded mouse lymph node tissue with Rabbit anti-ZAP70 antibody (HA723344) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723344) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723344_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-ZAP70 antibody (HA723344) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723344) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723344_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded mouse liver tissue (negative) with Rabbit anti-ZAP70 antibody (HA723344) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723344) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723344_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded rat lymph node tissue with Rabbit anti-ZAP70 antibody (HA723344) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723344) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723344_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-ZAP70 antibody (HA723344) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723344) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723344_11.jpg Fig11: Immunohistochemical analysis of paraffin-embedded rat liver tissue (negative) with Rabbit anti-ZAP70 antibody (HA723344) at 1/1,000 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA723344) at 1/1,000 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
HA723344_12.jpg Fig12: Flow cytometric analysis of Ramos (left, negative) and Jurkat (right, positive) cells labeling ZAP70.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA723344, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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