HA751806

VAV1 Recombinant Rabbit Monoclonal Antibody [PSH21-73]

cat.: HA751806

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Monoclonal
Clone number:	PSH21-73

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4).
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 98 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human VAV1 aa 551-800.
Positive control:	Jurkat cell lysate, Ramos cell lysate, THP-1 cell lysate, RAW264.7 cell lysate, Mouse thymus tissue lysate, Mouse spleen tissue lysate, THP-1, RAW264.7, mouse liver tissue, mouse spleen tissue, rat liver tissue, rat spleen tissue.
Subcellular location:	Cell-cell junction, cytoplasm, cytosol, plasma membrane.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:25,000 1:100 1:200-1:1,000 1:1,000
Uniprot #:	SwissProt: P15498 Human \| P27870 Mouse \| P54100 Rat
Alternative names:	Oncogene vav p95Vav Proto-oncogene vav Protooncogene vav VAV 1 VAV 1 oncogene VAV Vav proto oncogene VAV_HUMAN VAV1

Images

Fig1: Western blot analysis of VAV1 on different lysates with Rabbit anti-VAV1 antibody (HA751806) at 1/25,000 dilution.

Lane 1: Jurkat cell lysate (15 µg/Lane)
Lane 2: Ramos cell lysate (15 µg/Lane)
Lane 3: A549 cell lysate (negative) (15 µg/Lane)
Lane 4: 293T cell lysate (negative) (15 µg/Lane)
Lane 5: HeLa cell lysate (negative) (15 µg/Lane)
Lane 6: MDA-MB-231 cell lysate (low expression) (15 µg/Lane)
Lane 7: THP-1 cell lysate (15 µg/Lane)
Lane 8: RAW264.7 cell lysate (15 µg/Lane)

Predicted band size: 98 kDa
Observed band size: 98 kDa

Exposure time: 24 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 (HA751806) at 1/25,000 dilution was used in primary antibody dilution (K1803) 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: Western blot analysis of VAV1 on different lysates with Rabbit anti-VAV1 antibody (HA751806) at 1/25,000 dilution.

Lane 1: Mouse thymus tissue lysate (15 µg/Lane)
Lane 2: Mouse spleen tissue lysate (15 µg/Lane)

Predicted band size: 98 kDa
Observed band size: 98 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 (HA751806) at 1/25,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig3: Immunocytochemistry analysis of THP-1 cells labeling VAV1 with Rabbit anti-VAV1 antibody (HA751806) at 1/100 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-VAV1 antibody (HA751806) at 1/100 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.

Fig4: Immunocytochemistry analysis of RAW264.7 cells labeling VAV1 with Rabbit anti-VAV1 antibody (HA751806) at 1/100 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-VAV1 antibody (HA751806) at 1/100 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.

	Fig5: Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-VAV1 antibody (HA751806) 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 (HA751806) 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.
	Fig6: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-VAV1 antibody (HA751806) 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 (HA751806) 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-VAV1 antibody (HA751806) 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 (HA751806) 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.

Fig8: Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-VAV1 antibody (HA751806) 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 (HA751806) 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.

Fig9: Flow cytometric analysis of THP-1 cells labeling VAV1.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA751806, 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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