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, FC, IP
Clonality: Monoclonal
Clone number: JF0963
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
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
  IP
1:1,000-1:2,000
1:50-1:100
1:1,000
1:5,000
1:1,000
Use at an assay dependent concentration.
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 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; 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 (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_2.jpg Fig2: Western blot analysis of VAMP8 on different lysates with Rabbit anti-VAMP8 antibody (ET1702-10) at 1/1,000 dilution.

Lane 1: Hela cell lysate (10 µg/Lane)
Lane 2: Mouse kidney tissue lysate (20 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 1/1,000 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: 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_4.jpg Fig4: 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_5.jpg Fig5: 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_6.jpg Fig6: 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_7.jpg Fig7: 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_8.jpg Fig8: 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).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.