ET1705-94

Vinculin Recombinant Rabbit Monoclonal Antibody [JM42-43]

cat.: ET1705-94

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IF-Tissue, IHC-P
Clonality:	Monoclonal
Clone number:	JM42-43

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: 124 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within human Vinculin aa 1,000-1,060.
Positive control:	HepG2 cell lysate, HeLa cell lysate, NIH/3T3 cell lysate, C6 cell lysate, mouse spleen tissue lysate, rat kidney tissue lysate, HeLa, human testis tissue.
Subcellular location:	Cell junction, Cell membrane, Cytoplasm, Cytoskeleton, Membrane.
Recommended Dilutions: WB IF-Cell IF-Tissue IHC-P	1:5,000-1:20,000 1:200-1:1,000 1:50-1:4,000 1:200-1:20,000
Uniprot #:	SwissProt: P18206 Human \| Q64727 Mouse \| P85972 Rat
Alternative names:	CMD1W CMH15 Epididymis luminal protein 114 HEL114 Metavinculin MV MVCL OTTHUMP00000019861 OTTHUMP00000019862 VCL VINC VINC_HUMAN Vinculin

Images

	Fig1: Western blot analysis of Vinculin on different lysates with Rabbit anti-Vinculin antibody (ET1705-94) at 1/20,000 dilution. Lane 1: HepG2 cell lysate (15 µg/Lane) Lane 2: HeLa cell lysate (15 µg/Lane) Lane 3: NIH/3T3 cell lysate (15 µg/Lane) Lane 4: C6 cell lysate (15 µg/Lane) Lane 5: Mouse spleen tissue lysate (20 µg/Lane) Lane 6: Rat kidney tissue lysate (20 µg/Lane) Predicted band size: 124 kDa Observed band size: 124 kDa Exposure time: 1 minute 21 seconds; 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 (ET1705-94) at 1/20,000 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: Immunocytochemistry analysis of HeLa cells labeling Vinculin with Rabbit anti-Vinculin antibody (ET1705-94) at 1/1,000 dilution. Cells were fixed in 100% precooled methanol for 5 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-Vinculin antibody (ET1705-94) at 1/1,000 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.
	Fig3: Flow cytometric analysis of HeLa cells labeling Vinculin. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1705-94, 1μg/mL) (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).

	Fig4: Immunohistochemical analysis of paraffin-embedded human testis tissue with Rabbit anti-Vinculin antibody (ET1705-94) 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 (ET1705-94) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-Vinculin antibody (ET1705-94) at 1/20,000 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 (ET1705-94) at 1/20,000 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.
	Fig6: Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Vinculin antibody (ET1705-94) at 1/20,000 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 (ET1705-94) at 1/20,000 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 colon tissue with Rabbit anti-Vinculin antibody (ET1705-94) at 1/20,000 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 (ET1705-94) at 1/20,000 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.
	Fig8: Immunohistochemical analysis of paraffin-embedded rat spleen tissue with Rabbit anti-Vinculin antibody (ET1705-94) at 1/20,000 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 (ET1705-94) at 1/20,000 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: Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Vinculin antibody (ET1705-94) at 1/20,000 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 (ET1705-94) at 1/20,000 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.

Fig10: Immunocytochemistry analysis of NIH/3T3 cells labeling Vinculin with Rabbit anti-Vinculin antibody (ET1705-94) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Vinculin antibody (ET1705-94) at 1/1250 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.

Fig11: Immunocytochemistry analysis of C6 cells labeling Vinculin with Rabbit anti-Vinculin antibody (ET1705-94) at 1/250 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 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-Vinculin antibody (ET1705-94) at 1/1250 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.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.