ET7107-55

CD3G Recombinant Rabbit Monoclonal Antibody [JB38-29]

cat.: ET7107-55

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse
Applications:	WB, IHC-P, FC, IP, IF-Cell
Clonality:	Monoclonal
Clone number:	JB38-29

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: 20 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human CD3G aa 133-182 / 182.
Positive control:	Jurkat cell lysate, MOLT-4 cell lysate, mouse thymus tissue lysate, human tonsil tissue, human spleen tissue, mouse spleen tissue, mouse thymus tissue, Jurkat.
Subcellular location:	Cell membrane.
Recommended Dilutions: WB IHC-P FC IP IF-Cell	1:2,000 1:50-1:800 1:1,000 Use at an assay dependent concentration. 1:50
Uniprot #:	SwissProt: P09693 Human \| P11942 Mouse
Alternative names:	CD3 gamma CD3-GAMMA CD3g CD3g antigen gamma polypeptide (TiT3 complex) CD3g antigen gamma polypeptide CD3g molecule gamma (CD3 TCR complex) CD3g molecule gamma CD3G_HUMAN FLJ17620 FLJ17664 FLJ79544 FLJ94613 MGC138597 T cell antigen receptor complex gamma subunit of T3 T-cell receptor T3 gamma chain T-cell surface glycoprotein CD3 gamma chain T3G

Images

	Fig1: Western blot analysis of CD3G on different lysates with Rabbit anti-CD3G antibody (ET7107-55) at 1/2,000 dilution. Lane 1: Jurkat cell lysate Lane 2: MOLT-4 cell lysate Lane 3: Mouse thymus tissue lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 20 kDa Observed band size: 17/20 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 (ET7107-55) 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.
	Fig2: Immunohistochemical analysis of paraffin-embedded human tonsil tissue using anti-CD3G antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7107-55, 1/50) for 30 minutes 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.
	Fig3: Immunohistochemical analysis of paraffin-embedded human spleen tissue using anti-CD3G antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (ET7107-55, 1/50) for 30 minutes 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.

	Fig4: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-CD3G antibody (ET7107-55) at 1/800 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 (ET7107-55) at 1/800 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse thymus tissue with Rabbit anti-CD3G antibody (ET7107-55) at 1/800 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 (ET7107-55) at 1/800 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.
	Fig6: Immunocytochemistry analysis of Jurkat cells labeling CD3G with Rabbit anti-CD3G antibody (ET7107-55) at 1/50 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-CD3G antibody (ET7107-55) at 1/50 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 (M1305-2, 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.

Fig7: Flow cytometric analysis of Jurkat cells labeling CD3G.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET7107-55, 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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