ZAP70 Recombinant Rabbit Monoclonal Antibody [JU08-39]
cat.: ET1706-42
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human
Applications: WB, IF-Cell, IHC-P, FC, IP
Clonality: Monoclonal
Clone number: JU08-39
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.05% 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 570-619 / 619.
Positive control: Jurkat cell lysate, MOLT-4 cell lysate, HUT 102 cell lysate, human lymph node tissue, human spleen tissue, LOVO, SH-SY-5Y, Jurkat.
Subcellular location: Cytoplasm. Cell membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  IP
1:2,000
1:50-1:200
1:200
1:50-1:100
1:10-1:50
Uniprot #: SwissProt: P43403 Human
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
ET1706-42_1.jpg Fig1: Western blot analysis of ZAP70 on different lysates with Rabbit anti-ZAP70 antibody (ET1706-42) at 1/2,000 dilution.

Lane 1: Jurkat cell lysate (20 µg/Lane)
Lane 2: Ramos cell lysate (negative) (20 µg/Lane)
Lane 3: MOLT-4 cell lysate (20 µg/Lane)
Lane 4: Raji cell lysate (negative) (20 µg/Lane)
Lane 5: HUT 102 cell lysate (20 µg/Lane)

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

Exposure time: 40 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 (ET1706-42) 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.
ET1706-42_2.jpg Fig2: Immunohistochemical analysis of paraffin-embedded human lymph node tissue with Rabbit anti-ZAP70 antibody (ET1706-42) at 1/200 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 (ET1706-42) at 1/200 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.
ET1706-42_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-ZAP70 antibody (ET1706-42) at 1/200 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 (ET1706-42) at 1/200 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.
ET1706-42_4.jpg Fig4: Immunocytochemistry analysis of LOVO cells labeling ZAP70 with Rabbit anti-ZAP70 antibody (ET1706-42) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-ZAP70 antibody (ET1706-42) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET1706-42_5.jpg Fig5: Immunocytochemistry analysis of SH-SY-5Y cells labeling ZAP70 with Rabbit anti-ZAP70 antibody (ET1706-42) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-ZAP70 antibody (ET1706-42) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET1706-42_6.jpg Fig6: Flow cytometric analysis of ZAP70 was done on Jurkat cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1706-42, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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