Recombinant
RabMAb

Recombinant Anti-BST2/Tetherin antibody [EPR20202-150] (ab243230)

Overview

  • Product name

    Anti-BST2/Tetherin antibody [EPR20202-150]
    See all BST2/Tetherin primary antibodies
  • Description

    Rabbit monoclonal [EPR20202-150] to BST2/Tetherin
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, IP, Flow Cyt, WBmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human BST2/Tetherin aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: Q10589

  • Positive control

    • WB: HeLa whole cell lysate in the loading buffer containing DTT; K562 whole cell lysate in the loading buffer containing DTT. U937 whole cell lysate. ICC/IF: HeLa and U-937 cells. Flow Cyt: HeLa cells. IP: HeLa whole cell lysate.
  • General notes

     This product was previously labelled as BST2

     

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab243230 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
ICC/IF 1/100.
IP 1/40.
Flow Cyt 1/700.
WB 1/1000. Detects a band of approximately 35, 70 kDa (predicted molecular weight: 20 kDa).
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • Function

      May be involved in the sorting of secreted proteins (By similarity). May be involved in pre-B-cell growth. Antiretroviral defense protein, that blocks release of retrovirus from the cell surface. Depleted unpon HIV-1 infection by viral VPU protein through 20S proteasome degradation. Depleted upon infection by human Kaposi's sarcoma-associated herpesvirus (KSHV) through ubiquitination and subsequent degradation. May play a role in B-cell activation in rheumatoid arthritis.
    • Tissue specificity

      Predominantly expressed in liver, lung, heart and placenta. Lower levels in pancreas, kidney, skeletal muscle and brain. Overexpressed in multiple myeloma cells. Highly expressed during B-cell development, from pro-B precursors to plasma cells. Highly expressed on T-cells, monocytes, NK cells and dendritic cells (at protein level).
    • Sequence similarities

      Belongs to the tetherin family.
    • Domain

      The extracellular coiled coil domain is important for virus retention at the cell surface and prevention of virus spreading.
    • Post-translational
      modifications

      Monoubiquitinated by KSHV E3 ubiquitin-protein ligase K5, leading to its targeting to late endosomes and degradation.
    • Cellular localization

      Golgi apparatus > trans-Golgi network. Cell membrane. Cell membrane. Late endosome. Targeted to late endosomes upon KSHV infection and subsequent ubiquitination. Targeted to the trans-Golgi network by viral VPU protein.
    • Information by UniProt
    • Database links

    • Alternative names

      • Bone marrow stromal antigen 2 antibody
      • Bone marrow stromal cell antigen 2 antibody
      • Bone marrow stromal cell antigen antibody
      • BST 2 antibody
      • BST-2 antibody
      • BST2 antibody
      • BST2_HUMAN antibody
      • CD 317 antibody
      • CD317 antibody
      • CD317 antigen antibody
      • HM1.24 antigen antibody
      • NPC A 7 antibody
      • Tetherin antibody
      see all

    Images

    • BST2/Tetherin was immunoprecipitated from 0.35 mg HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab243230 at 1/40 dilution. Western blot was perfromed on the immunoprecipitate using ab243230 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366) was used afor detection at 1/5000 dilution.

      Lane 1: HeLa whole cell lysate 10 µg (input)
      Lane 2: ab243230 IP in HeLa whole cell lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab243230 in HeLa whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST
      Exposure time: 3 seconds.

    • Flow cytometric analysis of HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling BST2/Tetherin with ab243230 at 1/700 dilution (Red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U-937 (human histiocytic lymphoma cell line) cells labeling BST2/Tetherin with ab243230 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody (Green). Confocal image showing cytoplasmic staining in U-937 cells (PMID: 20529266).

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling BST2/Tetherin with ab243230 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody (Green). Confocal image showing cytoplasmic staining in HeLa cells (PMID: 20529266).

      The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Anti-BST2/Tetherin antibody [EPR20202-150] (ab243230) at 1/1000 dilution + U-937 (human histiocytic lymphoma cell line) whole cell lysate at 20 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 20 kDa


      Exposure time: 125 seconds


      Blocking/Diluting buffer and concentration: 5% NFDM/TBST

       

    • All lanes : Anti-BST2/Tetherin antibody [EPR20202-150] (ab243230) at 1/1000 dilution

      Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell), whole cell lysate in the loading buffer containing DTT
      Lane 2 : HeLa whole cell lysate in the loading buffer without DTT
      Lane 3 : K562 (human chronic myelogenous leukemia lymphoblast), whole cell lysate in the loading buffer containing
      Lane 4 : K562 whole cell lysate in the loading buffer without DTT

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 20 kDa
      Observed band size: 35,70 kDa
      why is the actual band size different from the predicted?


      Exposure time: 48 seconds


      Blocking/Diluting buffer and concentration: 5% NFDM/TBST

      The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID: 19196977; PMID: 19737401).

      BST2/Tetherin is a glycosylated protein, its calculated MW is 20 kDa, and the observed MW is 35 kDa, which is consistent with the literature.

      Both 35 and 70 kDa bands were detected under the reducing condition, whereas under the non-reducing condition, only the 70 kDa band was detected.

       

    References

    ab243230 has not yet been referenced specifically in any publications.

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