VAMP8 Recombinant Rabbit Monoclonal Antibody [JF0963]
cat.: ET1702-10
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IHC-P, IP, FC
Clonality: Monoclonal
Clone number: JF0963
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: 11 kDa
Isotype: IgG
Immunogen: Synthetic peptide wiithin Human VAMP8 aa1-29 / 100.
Positive control: Hela cell lysate, mouse kidney tissue lysate, U937 cell lysates, SW480, 293T, Hela, human kidney tissue, mouse kidney tissue, rat kidney tissue.
Subcellular location: Lysosome membrane, Early endosome membrane, Late endosome membrane, Cell membrane.
Recommended Dilutions:
  WB
  IF-Cell
  IF-Tissue
  IHC-P
  FC
1:1,000-1:2,000
1:50-1:100
1:1,000
1:5,000
1:50-1:100
Uniprot #: SwissProt: Q9BV40 Human | O70404 Mouse | Q9WUF4 Rat
Alternative names: EDB Endobrevin VAMP 8 VAMP-8 VAMP8 VAMP8_HUMAN Vesicle associated membrane protein 8 Vesicle-associated membrane protein 8
Images
ET1702-10_1.jpg Fig1: Western blot analysis of VAMP8 on different lysates with Rabbit anti-VAMP8 antibody (ET1702-10) at 1/1000 dilution.

Lane 1: Hela cell lysate
Lane 2: Mouse kidney tissue lysate (20 µg/Lane)

Lysates at 10 µg/Lane.

Predicted band size: 11 kDa
Observed band size: 11 kDa

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1702-10) at 1/1000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
ET1702-10_2.jpg Fig2: Western blot analysis of VAMP8 on U937 cell lysates with Rabbit anti-VAMP8 antibody (ET1702-10) at 1/500 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 11 kDa
Observed band size: 11 kDa

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1702-10) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:300,000 dilution was used for 1 hour at room temperature.
ET1702-10_3.jpg Fig3: Immunocytochemistry analysis of Hela cells labeling VAMP8 with Rabbit anti-VAMP8 antibody (ET1702-10) 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-VAMP8 antibody (ET1702-10) at 1/200 dilution in 2% negative goat serum 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.
ET1702-10_4.jpg Fig4: Immunocytochemistry analysis of SW480 cells labeling VAMP8 with Rabbit anti-VAMP8 antibody (ET1702-10) 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-VAMP8 antibody (ET1702-10) at 1/200 dilution in 2% negative goat serum 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.
ET1702-10_5.jpg Fig5: Immunocytochemistry analysis of 293T cells labeling VAMP8 with Rabbit anti-VAMP8 antibody (ET1702-10) 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-VAMP8 antibody (ET1702-10) at 1/200 dilution in 2% negative goat serum 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.
ET1702-10_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-VAMP8 antibody (ET1702-10) at 1/5,000 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 (ET1702-10) at 1/5,000 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.
ET1702-10_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-VAMP8 antibody (ET1702-10) at 1/5,000 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 (ET1702-10) at 1/5,000 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.
ET1702-10_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-VAMP8 antibody (ET1702-10) at 1/5,000 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 (ET1702-10) at 1/5,000 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.
ET1702-10_9.jpg Fig9: Flow cytometric analysis of Hela cells with VAMP8 antibody at 1/50 dilution (red) compared with an unlabelled control (cells without incubation with primary antibody; black). Alexa Fluor 488-conjugated goat anti rabbit IgG was used as the secondary antibody.
ET1702-10_10.jpg Fig10: Western blot analysis of VAMP8 on HL-60 cell/tissue lysates with Rabbit anti-VAMP8 antibody (ET1702-10) at 1/2,000 dilution.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 11 kDa
Observed band size: 15 kDa

Exposure time: 1 minutes 55 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 (ET1702-10) 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.
ET1702-10_11.jpg Fig11: Immunocytochemistry analysis of HL-60 cells labeling VAMP8 with Rabbit anti-VAMP8 antibody (ET1702-10) at 1/100 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-VAMP8 antibody (ET1702-10) 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 (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.
ET1702-10_12.jpg Fig12: Flow cytometric analysis of HL-60 cells labeling VAMP8.

Cells were fixed and permeabilized. Then stained with the primary antibody (ET1702-10, 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).
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