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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-VTI1B antibody [EPR15506(B)] (ab184170)

Reviews (1) Submit a question References (4)
Western blot - Anti-VTI1B antibody [EPR15506(B)] (ab184170)
  • Flow Cytometry (Intracellular) - Anti-VTI1B antibody [EPR15506(B)] (ab184170)
  • Western blot - Anti-VTI1B antibody [EPR15506(B)] (ab184170)
  • Western blot - Anti-VTI1B antibody [EPR15506(B)] (ab184170)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VTI1B antibody [EPR15506(B)] (ab184170)
  • Immunocytochemistry/ Immunofluorescence - Anti-VTI1B antibody [EPR15506(B)] (ab184170)
  • Anti-VTI1B antibody [EPR15506(B)] (ab184170)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR15506(B)] to VTI1B
  • Suitable for: IHC-P, WB, ICC/IF, Flow Cyt (Intra)
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-VTI1B antibody [EPR15506(B)]
    See all VTI1B primary antibodies

  • Description

    Rabbit monoclonal [EPR15506(B)] to VTI1B

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WB, ICC/IF, Flow Cyt (Intra)more details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Mouse, Rat

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: MCF7 and HeLa cell lysates; Human fetal lung and fetal liver lysates. IHC-P: Human ovarian carcinoma tissue. ICC/IF: MCF7 cells.

  • General notes

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    pH: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR15506(B)

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab184170 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000 - 1/10000. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa).
ICC/IF
1/50.
Flow Cyt (Intra)
1/170.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Notes
IHC-P
1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB
1/1000 - 1/10000. Detects a band of approximately 27 kDa (predicted molecular weight: 27 kDa).
ICC/IF
1/50.
Flow Cyt (Intra)
1/170.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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.

  • Tissue specificity

    Expressed in all tissues examined.

  • Sequence similarities

    Belongs to the VTI1 family.

  • Cellular localization

    Endomembrane system. Cytoplasmic granule.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • 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
    • Vti1 rp1 antibody
    • VTI1-LIKE antibody
    • Vti1-rp1 antibody
    • VTI1B antibody
    • VTI1B_HUMAN antibody
    • VTI1L antibody
    • VTI1L1 antibody
    • VTI2 antibody
    see all

Images

  • Western blot - Anti-VTI1B antibody [EPR15506(B)] (ab184170)
    Western blot - Anti-VTI1B antibody [EPR15506(B)] (ab184170)
    All lanes : Anti-VTI1B antibody [EPR15506(B)] (ab184170) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : VTI1B knockout HeLa cell lysate
    Lane 3 : MCF7 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 27 kDa
    Observed band size: 28 kDa why is the actual band size different from the predicted?



    Lanes 1-3: Merged signal (red and green). Green - ab184170 observed at 28 kDa. Red - loading control ab8245 observed at 36 kDa. 

     ab184170 Anti-VTI1B antibody [EPR15506(B)]  was shown to specifically react with VTI1B in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265483 (knockout cell lysate ab257797) was used.  Wild-type and VTI1B knockout samples were subjected to SDS-PAGE.  ab184170 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively.  Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Flow Cytometry (Intracellular) - Anti-VTI1B antibody [EPR15506(B)] (ab184170)
    Flow Cytometry (Intracellular) - Anti-VTI1B antibody [EPR15506(B)] (ab184170)

    Intracellular flow cytometric analysis of Hela cells (2% paraformaldehyde-fixed) labeling VTI1B with ab184170 at 1/170 dilution (red) or a rabbit IgG (negative) (green), followed by Goat anti rabbit IgG (FITC) secondary at 1/150 dilution. 

     

     

  • Western blot - Anti-VTI1B antibody [EPR15506(B)] (ab184170)
    Western blot - Anti-VTI1B antibody [EPR15506(B)] (ab184170)
    All lanes : Anti-VTI1B antibody [EPR15506(B)] (ab184170) at 1/2000 dilution

    Lane 1 : MCF7 cell lysate
    Lane 2 : HeLa cell lysate
    Lane 3 : Human fetal liver lysate
    Lane 4 : Human fetal lung lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 27 kDa

  • Western blot - Anti-VTI1B antibody [EPR15506(B)] (ab184170)
    Western blot - Anti-VTI1B antibody [EPR15506(B)] (ab184170)
    Anti-VTI1B antibody [EPR15506(B)] (ab184170) at 1/10000 dilution + MCF7 cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

    Predicted band size: 27 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VTI1B antibody [EPR15506(B)] (ab184170)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-VTI1B antibody [EPR15506(B)] (ab184170)

    Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma labeling VTI1B with ab184170 at 1:100 diluton and HRP polymer.  Counterstained with Hematoxylin.

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-VTI1B antibody [EPR15506(B)] (ab184170)
    Immunocytochemistry/ Immunofluorescence - Anti-VTI1B antibody [EPR15506(B)] (ab184170)

    Immunofluorescent analysis of MCF7 cells (paraformaldehyde-fixed, 4%) labeling VTI1B with ab184170 at 1/50 dilution followed by Goat anti rabbit IgG (AlexaFluor® 488) secondary at 1/200 dilution and counter-stained with DAPI (blue).

  • Anti-VTI1B antibody [EPR15506(B)] (ab184170)
    Anti-VTI1B antibody [EPR15506(B)] (ab184170)

References (4)

Publishing research using ab184170? Please let us know so that we can cite the reference in this datasheet.

ab184170 has been referenced in 4 publications.

  • Coukos R  et al. An engineered transcriptional reporter of protein localization identifies regulators of mitochondrial and ER membrane protein trafficking in high-throughput CRISPRi screens. Elife 10:N/A (2021). PubMed: 34414886
  • Margiotta A  et al. Invariant chain regulates endosomal fusion and maturation through an interaction with the SNARE Vti1b. J Cell Sci 133:N/A (2020). PubMed: 32907852
  • Abed M  et al. The Gag protein PEG10 binds to RNA and regulates trophoblast stem cell lineage specification. PLoS One 14:e0214110 (2019). PubMed: 30951545
  • Lesteberg K  et al. Recycling endosomes in human cytotoxic T lymphocytes constitute an auxiliary intracellular trafficking pathway for newly synthesized perforin. Immunol Res 65:1031-1045 (2017). Flow Cyt ; Human . PubMed: 28822075

Customer reviews and Q&As

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Immunocytochemistry/ Immunofluorescence abreview for Anti-VTI1B antibody [EPR15506(B)]

 Inconclusive
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (Mouse cells transfected with HUMAN GFP-Vti1b)
Permeabilization
No
Specification
Mouse cells transfected with HUMAN GFP-Vti1b
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 23°C
Fixative
Methanol
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Apr 04 2022

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