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VAMP8 Recombinant Rabbit Monoclonal Antibody [JF0963]

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Specification

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Catalog# ET1702-10

VAMP8 Recombinant Rabbit Monoclonal Antibody [JF0963]

Application

  • WB

  • IF-Cell

  • IF-Tissue

  • IHC-P

  • FC

  • IP

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

This product has been cited in peer reviewed publications, see list HERE

Overview

Product Name

VAMP8 Recombinant Rabbit Monoclonal Antibody [JF0963]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Synthetic peptide wiithin Human VAMP8 aa1-29 / 100.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IF-Tissue, IHC-P, FC, IP

Molecular Weight

Predicted band size: 11 kDa

Positive Control

Hela cell lysate, mouse kidney tissue lysate, U937 cell lysates, SW480, 293T, Hela, human kidney tissue, mouse kidney tissue, rat kidney tissue.

Conjugation

unconjugated

Clone Number

JF0963

RRID

AB_3070273

Product Features

Form

Liquid

Concentration

Storage Instructions

Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000-1:2,000

  • IF-Cell

  • 1:50-1:100

  • IF-Tissue

  • 1:1,000

  • IHC-P

  • 1:5,000

  • FC

  • 1:1,000

  • IP

  • Use at an assay dependent concentration.

Target

Function

Vesicle-associated membrane protein 8 is a protein that in humans is encoded by the VAMP8 gene.Synaptobrevins/VAMPs, syntaxins, and the 25-kD synaptosomal-associated protein SNAP25 are the main components of a protein complex involved in the docking and/or fusion of synaptic vesicles with the presynaptic membrane. The protein encoded by this gene is a member of the vesicle-associated membrane protein (VAMP)/synaptobrevin family. It is associated with the perinuclear vesicular structures of the early endocytic compartment. It has been found that VAMP8 interacts specifically with the soluble NSF-attachment protein (alpha-SNAP), most likely through an VAMP8-containing SNARE complex.Phosphorylation of VAMP8 inside the conserved SNARE-domain can suppress vesicle fusion.

Background References

1. Xie X et al. Deep vein thrombosis is accurately predicted by comprehensive analysis of the levels of microRNA-96 and plasma D-dimer. Exp Ther Med 12:1896-1900 (2016).

2. Pirooz SD et al. UVRAG is required for virus entry through combinatorial interaction with the class C-Vps complex and SNAREs. Proc Natl Acad Sci U S A 111:2716-21 (2014).

Sequence Similarity

Belongs to the synaptobrevin family.

Tissue Specificity

Platelets.

Subcellular Location

Lysosome membrane, Early endosome membrane, Late endosome membrane, Cell membrane.

UNIPROT

SwissProt: Q9BV40 Human

SwissProt: O70404 Mouse

SwissProt: Q9WUF4 Rat

Synonyms

EDB antibody

Endobrevin antibody

VAMP 8 antibody

VAMP-8 antibody

VAMP8 antibody

VAMP8_HUMAN antibody

Vesicle associated membrane protein 8 antibody

Vesicle-associated membrane protein 8 antibody

Images

  • Western blot analysis of VAMP8 on HL-60 cell/tissue lysates with Rabbit anti-VAMP8 antibody (<a href="/products/ET1702-10" style="font-weight: bold;text-decoration: underline;">ET1702-10</a>) at 1/2,000 dilution.<br /><br />Lysates/proteins at 20 µg/Lane.<br /><br />Predicted band size: 11 kDa<br />Observed band size: 15 kDa<br /><br />Exposure time: 1 minutes 55 seconds; ECL: K1801;<br />4-20% SDS-PAGE gel.<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1702-10" style="font-weight: bold;text-decoration: underline;">ET1702-10</a>) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/50,000 dilution was used for 1 hour at room temperature.

    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.

  • Western blot analysis of VAMP8 on different lysates with Rabbit anti-VAMP8 antibody (<a href="/products/ET1702-10" style="font-weight: bold;text-decoration: underline;">ET1702-10</a>) at 1/1,000 dilution.<br /><br />Lane 1: Hela cell lysate (10 µg/Lane)<br />Lane 2: Mouse kidney tissue lysate (20 µg/Lane)<br /><br />Predicted band size: 11 kDa<br />Observed band size: 11 kDa<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1702-10" style="font-weight: bold;text-decoration: underline;">ET1702-10</a>) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/300,000 dilution was used for 1 hour at room temperature.

    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.

  • Western blot analysis of VAMP8 on U937 cell lysates with Rabbit anti-VAMP8 antibody (<a href="/products/ET1702-10" style="font-weight: bold;text-decoration: underline;">ET1702-10</a>) at 1/500 dilution.<br /><br />Lysates/proteins at 10 µg/Lane.<br /><br />Predicted band size: 11 kDa<br />Observed band size: 11 kDa<br /><br />Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (<a href="/products/ET1702-10" style="font-weight: bold;text-decoration: underline;">ET1702-10</a>) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (<a href="/products/HA1001" style="font-weight: bold;text-decoration: underline;">HA1001</a>) at 1/300,000 dilution was used for 1 hour at room temperature.

    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.

  • Immunocytochemistry analysis of HL-60 cells labeling VAMP8 with Rabbit anti-VAMP8 antibody (<a href="/products/ET1702-10" style="font-weight: bold;text-decoration: underline;">ET1702-10</a>) at 1/100 dilution.<br /><br />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 (<a href="/products/ET1702-10" style="font-weight: bold;text-decoration: underline;">ET1702-10</a>) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor&trade; 488, <a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) 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.<br /><br />Beta tubulin (<a href="/products/M1305-2" style="font-weight: bold;text-decoration: underline;">M1305-2</a>, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor&trade; 594, <a href="/products/HA1126" style="font-weight: bold;text-decoration: underline;">HA1126</a>) was used as the secondary antibody at 1/1,000 dilution.

    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.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue with Rabbit anti-VAMP8 antibody (<a href="/products/ET1702-10" style="font-weight: bold;text-decoration: underline;">ET1702-10</a>) at 1/5,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1702-10" style="font-weight: bold;text-decoration: underline;">ET1702-10</a>) 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.

    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.

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Rabbit anti-VAMP8 antibody (<a href="/products/ET1702-10" style="font-weight: bold;text-decoration: underline;">ET1702-10</a>) at 1/5,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1702-10" style="font-weight: bold;text-decoration: underline;">ET1702-10</a>) 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.

    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.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Rabbit anti-VAMP8 antibody (<a href="/products/ET1702-10" style="font-weight: bold;text-decoration: underline;">ET1702-10</a>) at 1/5,000 dilution.<br /><br />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 ddH<sub>2</sub>O and PBS, and then probed with the primary antibody (<a href="/products/ET1702-10" style="font-weight: bold;text-decoration: underline;">ET1702-10</a>) 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.

    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.

  • Flow cytometric analysis of HL-60 cells labeling VAMP8.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/ET1702-10" style="font-weight: bold;text-decoration: underline;">ET1702-10</a>, 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&trade; 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (<a href="/products/HA1121" style="font-weight: bold;text-decoration: underline;">HA1121</a>) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

    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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