Recombinant
RabMAb

Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853)

Overview

  • Product name
    Anti-beta 2 Microglobulin antibody [EP2978Y]
    See all beta 2 Microglobulin primary antibodies
  • Description
    Rabbit monoclonal [EP2978Y] to beta 2 Microglobulin
  • Tested applications
    Suitable for: ICC/IF, WB, IP, Flow Cytmore details
    Unsuitable for: IHC-P
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) within Human beta 2 Microglobulin (C terminal). The exact sequence is proprietary.
    (Peptide available as ab189068)

  • Positive control
    • WB: Raji, U937 and HeLa cell lysates and human bone marrow, rat and mouse kidney tissue lysates. ICC/IF: HeLa cells. IP: Raji cell lysate. FC: HeLa cells
  • General notes

     

     

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab75853 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 - 1/250.
WB 1/5000. Predicted molecular weight: 14 kDa.Can be blocked with beta 2 Microglobulin peptide (ab189068).

For unpurified use at 1/10000 - 1/50000.

The unpurified antibody has been found to work at dilutions up to 1/200,000 in WB.

IP 1/20 - 1/40.
Flow Cyt 1/20 - 1/60.
  • Application notes
    Is unsuitable for IHC-P.
  • Target

    • Function
      Component of the class I major histocompatibility complex (MHC). Involved in the presentation of peptide antigens to the immune system.
    • Involvement in disease
      Defects in B2M are the cause of hypercatabolic hypoproteinemia (HYCATHYP) [MIM:241600]. Affected individuals show marked reduction in serum concentrations of immunoglobulin and albumin, probably due to rapid degradation.
      Note=Beta-2-microglobulin may adopt the fibrillar configuration of amyloid in certain pathologic states. The capacity to assemble into amyloid fibrils is concentration dependent. Persistently high beta(2)-microglobulin serum levels lead to amyloidosis in patients on long-term hemodialysis.
    • Sequence similarities
      Belongs to the beta-2-microglobulin family.
      Contains 1 Ig-like C1-type (immunoglobulin-like) domain.
    • Post-translational
      modifications
      Glycation of Ile-21 is observed in long-term hemodialysis patients.
    • Cellular localization
      Secreted. Detected in serum and urine.
    • Information by UniProt
    • Database links
    • Alternative names
      • B2M antibody
      • B2MG_HUMAN antibody
      • Beta 2 microglobin antibody
      • Beta 2 microglobulin antibody
      • Beta 2 microglobulin precursor antibody
      • Beta chain of mhc class 1 proteins antibody
      • Beta chain of MHC class I molecules antibody
      • Beta-2-microglobulin form pI 5.3 antibody
      • CDABP0092 antibody
      • Hdcma22p antibody
      • IMD43 antibody
      see all

    Anti-beta 2 Microglobulin antibody [EP2978Y] images

    • All lanes : Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853) at 1/5000 dilution (purified)

      Lane 1 : U937 whole cell lysate
      Lane 2 : HeLa whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 14 kDa
      Observed band size : 14 kDa

      Blocking and dilution buffer: 5% NFDM /TBST.

    • Immunocytochemistry/Immunofluorescence analysis of HeLa cells labelling beta 2 Microglobulin with purified ab75853 at 1/100. Cells were fixed with 100% methanol and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

      Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

      Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

    • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling beta 2 Microglobulin with purified ab75853 at 1/20 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    • All lanes : Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853) at 1/5000 dilution (purified)

      Lane 1 : Mouse kidney tissue lysate
      Lane 2 : Rat kidney tissue lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size : 14 kDa
      Observed band size : 14 kDa

      Blocking and dilution buffer: 5% NFDM /TBST.

    • ab75853 (purified) at 1/20 immunoprecipitating beta 2 Microglobulin in Raji whole cell lysate.

      Lane 1 (input): Raji whole cell lysate (10µg)

      Lane 2 (+): ab75853 + Raji whole cell lysate (10µg).

      Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab75853 in Raji whole cell lysate.

      For western blotting, ab131366 VeriBlot for IP (HRP) was used as the secondary antibody (1/10000).

      Blocking buffer and concentration: 5% NFDM/TBST.

      Diluting buffer and concentration: 5% NFDM /TBST.

    • All lanes : Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853) at 1/200000 dilution (unpurified)

      Lane 1 : Raji cell lysate
      Lane 2 : U937 cell lysate
      Lane 3 : HeLa cell lysate
      Lane 4 : Human bone marrow lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

      Predicted band size : 14 kDa
      Observed band size : 14 kDa

    References for Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853)

    This product has been referenced in:
    • Melleby AO  et al. The Heparan Sulfate Proteoglycan Glypican-6 Is Upregulated in the Failing Heart, and Regulates Cardiomyocyte Growth through ERK1/2 Signaling. PLoS One 11:e0165079 (2016). WB ; Mouse, Human . Read more (PubMed: 27768722) »
    • Rebo J  et al. A single heterochronic blood exchange reveals rapid inhibition of multiple tissues by old blood. Nat Commun 7:13363 (2016). WB ; Mouse . Read more (PubMed: 27874859) »

    See all 10 Publications for this product

    Product Wall

    Application
    Immunohistochemistry (Frozen sections)
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
    Sample
    Mouse Tissue sections (Brain)
    Specification
    Brain
    Permeabilization
    Yes - 0.1M PBS with 3% TritonX100
    Fixative
    Paraformaldehyde
    Username

    Dr. Ruma Raha-Chowdhury

    Verified customer

    Submitted Aug 14 2014

    Application
    Western blot
    Loading amount
    10 µg
    Gel Running Conditions
    Reduced Denaturing (12% Tris-HCl)
    Sample
    Rabbit Cell lysate - whole cell (MDCK cell lysate transfected with human, mouse and)
    Specification
    MDCK cell lysate transfected with human, mouse and
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Jonathan

    Verified customer

    Submitted Aug 13 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Loading amount
    10 µg
    Gel Running Conditions
    Reduced Denaturing (12% Tris-HCl)
    Sample
    Mouse Cell lysate - whole cell (MDCK cell lysate transfected with human, mouse and)
    Specification
    MDCK cell lysate transfected with human, mouse and
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Jonathan

    Verified customer

    Submitted Aug 13 2013

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Loading amount
    5 µg
    Gel Running Conditions
    Reduced Denaturing (12% Tris-HCl)
    Sample
    Human Cell lysate - whole cell (MDCK cell lysate transfected with human, mouse and)
    Specification
    MDCK cell lysate transfected with human, mouse and
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
    Username

    Jonathan

    Verified customer

    Submitted Jul 30 2013

    In review of the immunogen sequence used for creating ab75853 I see this sequence share only 69.2% identity with rabbit beta-2-microglobulin. Generally we do not expect reactivity below 80%. Overall I have found that the sequence homologies of human...

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    Thank you for contacting us. A cy5 conjugated antibody against rabbit such as ab97077 or an Alexa 647 antibody such as ab150075 would be good choices for you.

    I hope this information is helpful to you. Please do not hesitate to contact us if y...

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    Thank you for your response and for participating in our Abreview Testing program. The code in brackets in the subject heading is a reference number for these correspondences. Should you wish to test another of the products, referencing this number wou...

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    Thank you for your inquiry.

    According to my information the mouse strain DBA/2NCrl has the haplotype H2d. Please encourage your customer strongly to double check this for the strain they mentioned (DBA II N strain).

    We do not have a...

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    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Glioblastoma Initiating Cells (Primary cells))
    Loading amount
    30 µg
    Specification
    Glioblastoma Initiating Cells (Primary cells)
    Gel Running Conditions
    Reduced Denaturing (12.5)
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 20°C
    Username

    Abcam user community

    Verified customer

    Submitted Nov 23 2011

    Thank you for your inquiry. When performing western blots on immunoprecipitated samples, it is recommended to use the primary antibody 10-25 times more concentrated than that used in western blots on lysates. This is the reason why an antibody used...

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