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VTI1B Recombinant Rabbit Monoclonal Antibody [PSH06-40]

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Specification

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Catalog# HA722675

VTI1B Recombinant Rabbit Monoclonal Antibody [PSH06-40]

Application

  • WB

  • IF-Cell

  • IHC-P

  • FC

Reactivity

  • Human

  • Mouse

  • Rat

BSA and Azide free
Conjugation

  • unconjugated

Overview

Product Name

VTI1B Recombinant Rabbit Monoclonal Antibody [PSH06-40]

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within human VTI1B aa 1-232.

Species Reactivity

Human, Mouse, Rat

Validated Applications

WB, IF-Cell, IHC-P, FC

Target Molecular Weight

Predicted band size: 27 kDa

Positive Control

HeLa cell lysate, MCF7 cell lysate, U-2 OS cell lysate, U-87 MG cell lysate, SH-SY5Y cell lysate, C2C12 cell lysate, PC-12 cell lysate, C6 cell lysate, Rat brain tissue lysate, Rat kidney tissue lysate, PC-12, human brain tissue, human kidney tissue, rat brain tissue.

Conjugation

unconjugated

Clone Number

PSH06-40

Product Features

Form

Liquid

Concentration

1 mg/mL.(The concentration of this product may be batch-dependent)

Storage Instructions

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

Storage Buffer

PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • WB

  • 1:1,000

  • IF-Cell

  • 1:100

  • IHC-P

  • 1:2,000

  • FC

  • 1:1,000

Target

Function

V-SNARE that mediates vesicle transport pathways through interactions with t-SNAREs on the target membrane. These interactions are proposed to mediate aspects of the specificity of vesicle trafficking and to promote fusion of the lipid bilayers. May be concerned with increased secretion of cytokines associated with cellular senescence. Enables SNARE binding activity and chloride channel inhibitor activity. Involved in regulation of protein localization to plasma membrane. Located in several cellular components, including endosome membrane; lysosomal membrane; and perinuclear region of cytoplasm.

Background References

1. Bollmann C et al. Primary neurons lacking the SNAREs vti1a and vti1b show altered neuronal development. Neural Dev. 2022 Nov

2. Chou HY et al. PTPN9-mediated dephosphorylation of VTI1B promotes ATG16L1 precursor fusion and autophagosome formation. Autophagy. 2021 Oct

Subcellular Location

Early endosome membrane, Late endosome membrane, Lysosome membrane, Cytoplasmic granule, Recycling endosome membrane.

UNIPROT

SwissProt: Q9UEU0 Human

SwissProt: O88384 Mouse

SwissProt: P58200 Rat

Synonyms

HGNC:17793 antibody

V SNARE antibody

v-SNARE antibody

Vesicle associated soluble NSF attachment protein receptor antibody

Vesicle transport through interaction with t SNAREs 1B antibody

Vesicle transport through interaction with t SNAREs homolog 1B (yeast) antibody

Vesicle transport through interaction with t-SNAREs homolog 1B antibody

Vesicle transport v-SNARE protein Vti1-like 1 antibody

VTI1 antibody

VTI1 like antibody

Expand

Images

  • Western blot analysis of VTI1B on different lysates with Rabbit anti-VTI1B antibody (<a href="/products/HA722675" style="font-weight: bold;text-decoration: underline;">HA722675</a>) at 1/1,000 dilution.<br /><br />Lane 1: HeLa cell lysate (20 µg/Lane)<br />Lane 2: MCF7 cell lysate (20 µg/Lane)<br />Lane 3: U-2 OS cell lysate (20 µg/Lane)<br />Lane 4: U-87 MG cell lysate (20 µg/Lane)<br />Lane 5: SH-SY5Y cell lysate (20 µg/Lane)<br />Lane 6: C2C12 cell lysate (20 µg/Lane)<br />Lane 7: PC-12 cell lysate (20 µg/Lane)<br />Lane 8: C6 cell lysate (20 µg/Lane)<br />Lane 9: Rat brain tissue lysate (40 µg/Lane)<br />Lane 10: Rat kidney tissue lysate (40 µg/Lane)<br /><br />Predicted band size: 27 kDa<br />Observed band size: 27 kDa<br /><br />Exposure time: 3 minutes; ECL: K1801;<br /><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/HA722675" style="font-weight: bold;text-decoration: underline;">HA722675</a>) at 1/1,000 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 VTI1B on different lysates with Rabbit anti-VTI1B antibody (HA722675) at 1/1,000 dilution.

    Lane 1: HeLa cell lysate (20 µg/Lane)
    Lane 2: MCF7 cell lysate (20 µg/Lane)
    Lane 3: U-2 OS cell lysate (20 µg/Lane)
    Lane 4: U-87 MG cell lysate (20 µg/Lane)
    Lane 5: SH-SY5Y cell lysate (20 µg/Lane)
    Lane 6: C2C12 cell lysate (20 µg/Lane)
    Lane 7: PC-12 cell lysate (20 µg/Lane)
    Lane 8: C6 cell lysate (20 µg/Lane)
    Lane 9: Rat brain tissue lysate (40 µg/Lane)
    Lane 10: Rat kidney tissue lysate (40 µg/Lane)

    Predicted band size: 27 kDa
    Observed band size: 27 kDa

    Exposure time: 3 minutes; 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 (HA722675) at 1/1,000 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.

  • Immunocytochemistry analysis of PC-12 cells labeling VTI1B with Rabbit anti-VTI1B antibody (<a href="/products/HA722675" style="font-weight: bold;text-decoration: underline;">HA722675</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-VTI1B antibody (<a href="/products/HA722675" style="font-weight: bold;text-decoration: underline;">HA722675</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 PC-12 cells labeling VTI1B with Rabbit anti-VTI1B antibody (HA722675) 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-VTI1B antibody (HA722675) 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 brain tissue with Rabbit anti-VTI1B antibody (<a href="/products/HA722675" style="font-weight: bold;text-decoration: underline;">HA722675</a>) at 1/2,000 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/HA722675" style="font-weight: bold;text-decoration: underline;">HA722675</a>) at 1/2,000 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 brain tissue with Rabbit anti-VTI1B antibody (HA722675) at 1/2,000 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 (HA722675) at 1/2,000 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-VTI1B antibody (<a href="/products/HA722675" style="font-weight: bold;text-decoration: underline;">HA722675</a>) at 1/2,000 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/HA722675" style="font-weight: bold;text-decoration: underline;">HA722675</a>) at 1/2,000 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-VTI1B antibody (HA722675) at 1/2,000 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 (HA722675) at 1/2,000 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 brain tissue with Rabbit anti-VTI1B antibody (<a href="/products/HA722675" style="font-weight: bold;text-decoration: underline;">HA722675</a>) at 1/2,000 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/HA722675" style="font-weight: bold;text-decoration: underline;">HA722675</a>) at 1/2,000 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 brain tissue with Rabbit anti-VTI1B antibody (HA722675) at 1/2,000 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 (HA722675) at 1/2,000 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 PC-12 cells labeling VTI1B.<br /><br />Cells were fixed and permeabilized. Then stained with the primary antibody (<a href="/products/HA722675" style="font-weight: bold;text-decoration: underline;">HA722675</a>, 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&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 PC-12 cells labeling VTI1B.

    Cells were fixed and permeabilized. Then stained with the primary antibody (HA722675, 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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