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Rabbit Recombinant Monoclonal beta 2 Microglobulin antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 57 publications.


Images

Western blot - Anti-beta 2 Microglobulin antibody [EP2978Y] (AB75853), expandable thumbnail
  • Western blot - Anti-beta 2 Microglobulin antibody [EP2978Y] (AB75853), expandable thumbnail
  • Immunoprecipitation - Anti-beta 2 Microglobulin antibody [EP2978Y] (AB75853), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-beta 2 Microglobulin antibody [EP2978Y] (AB75853), expandable thumbnail
  • Western blot - Anti-beta 2 Microglobulin antibody [EP2978Y] (AB75853), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IPWBIHC-PICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Not recommended
Tested
Tested
Mouse
Expected
Tested
Not recommended
Expected
Tested
Rat
Expected
Tested
Not recommended
Expected
Expected

Tested
Tested

Species

Human

Dilution info
1/20 - 1/40
Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Mouse

Dilution info
1/5000
Notes

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.

Species

Rat

Dilution info
1/5000
Notes

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.

Species

Human

Dilution info
1/5000
Notes

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.

Not recommended
Not recommended

Species

Mouse, Rat, Human

Dilution info

-

Notes

-

Tested
Tested

Species

Human

Dilution info
1/100 - 1/250
Notes

-

Expected
Expected

Species

Mouse, Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

Species

Human

Dilution info
1/20 - 1/60
Notes

-

Species

Mouse

Dilution info
1/20 - 1/60
Notes

-

Expected
Expected

Species

Rat

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

15 products for Alternative Product

Target data

Function

Component of the class I major histocompatibility complex (MHC). Involved in the presentation of peptide antigens to the immune system. Exogenously applied M.tuberculosis EsxA or EsxA-EsxB (or EsxA expressed in host) binds B2M and decreases its export to the cell surface (total protein levels do not change), probably leading to defects in class I antigen presentation (PubMed:25356553).

Alternative names

CDABP0092, HDCMA22P, B2M, Beta-2-microglobulin

Recommended products

Rabbit Recombinant Monoclonal beta 2 Microglobulin antibody. Suitable for IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Human, Mouse, Rat samples. Cited in 57 publications.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EP2978Y

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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.

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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Beta-2-Microglobulin (B2M) is a component of the class I major histocompatibility complex (MHC I) and plays an important role in presenting peptides to the immune system. B2M weighs approximately 11.8 kDa and is found abundantly in all nucleated cells. It has alternate names such as B2 microglobulin or beta-2-microglobulin. This protein is present in the cell membrane as a part of the MHC I which is important for immune surveillance. Additionally B2M is detectable in various biological fluids including serum and its levels can reflect physiological and pathological states.

Biological function summary

Beta-2-microglobulin is important for the stability and transport of MHC class I molecules to the cell surface. As part of the MHC class I complex B2M assists in binding peptides allowing immune cells to identify and target pathogen-infected cells. Without B2M the MHC class I molecules are not properly expressed on the cell surface disrupting immune recognition. In laboratory settings researchers often use anti-beta-2-microglobulin antibodies to investigate its role in MHC class I function.

Pathways

Beta-2-microglobulin interacts significantly with the immune system most notably in the antigen processing and presentation pathway. It works alongside proteins such as the heavy chain of MHC class I. B2M is important in the pathway that involves the transport of antigens to the endoplasmic reticulum where they are loaded onto MHC class I molecules for inspection by cytotoxic T cells. Another related pathway is the tapasin-mediated processing of antigen peptides highlighting the indispensable role of B2M in immune response regulation.

Associated diseases and disorders

Beta-2-microglobulin is associated with conditions such as beta-2-microglobulin amyloidosis and certain lymphoproliferative disorders. Elevated levels of B2M in serum serve as a marker for diseases like multiple myeloma where the protein level correlates with disease severity. B2M-related amyloidosis frequently occurs in patients undergoing long-term dialysis where amyloid deposits accumulate in tissues. Linking B2M to immune system dysfunction studies have shown interactions with other proteins including components of the immune system like HLA-A and HLA-B highlighting B2M's relevance in diagnosing and understanding these conditions.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

10 product images

  • Western blot - Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853), expandable thumbnail

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

    Lanes 1-4: Merged signal (red and green). Green - ab75853 observed at 14 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.

    ab75853 Anti-beta 2 Microglobulin antibody [EP2978Y] was shown to specifically react with beta 2 Microglobulin in wild-type HepG2 cells. Loss of signal was observed when knockout cell line Human B2M (beta 2 Microglobulin) knockout Hep G2 cell line ab262325 (knockout cell lysate Human B2M (beta 2 Microglobulin) knockout Hep G2 cell lysate ab256846) was used. Wild-type and beta 2 Microglobulin knockout samples were subjected to SDS-PAGE. ab75853 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.

    All lanes: Western blot - Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853) at 1/1000 dilution

    Lane 1: Wild-type HepG2 cell lysate at 20 µg

    Lane 2: B2M knockout HepG2 cell lysate at 20 µg

    Lane 2: Western blot - Human B2M (beta 2 Microglobulin) knockout Hep G2 cell line (Human B2M (beta 2 Microglobulin) knockout Hep G2 cell line ab262325)

    Lane 3: HeLa cell lysate at 20 µg

    Lane 4: Jurkat cell lysate at 20 µg

    Secondary

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

    Predicted band size: 14 kDa

    Observed band size: 14 kDa

    This data was developed using the same antibody clone in a different buffer formulation (ab75853).

    Lanes 1-4: Merged signal (red and green). Green - ab75853 observed at 14 kDa. Red - loading control Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 36 kDa.

    ab75853 Anti-beta 2 Microglobulin antibody [EP2978Y] was shown to specifically react with beta 2 Microglobulin in wild-type HepG2 cells. Loss of signal was observed when knockout cell line Human B2M (beta 2 Microglobulin) knockout Hep G2 cell line ab262325 (knockout cell lysate Human B2M (beta 2 Microglobulin) knockout Hep G2 cell lysate ab256846) was used. Wild-type and beta 2 Microglobulin knockout samples were subjected to SDS-PAGE. ab75853 and Anti-GAPDH antibody [6C5] - Loading Control (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (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.

  • Western blot - Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853), expandable thumbnail

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

    Western blot: Anti-beta 2 Microglobulin antibody [EP2978Y] staining at 1/5000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A]  (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab75853 was shown to bind specifically to beta 2 Microglobulin. A band was observed at 12 kDa in Wild-type A431 and HEK-293T cell lysates with no signal observed at this size in B2M (beta 2 Microglobulin) knockout A-431 cell line Human B2M (beta 2 Microglobulin) knockout A-431 cell line ab261893 (knockout cell lysate Human B2M (beta 2 Microglobulin) knockout A-431 cell lysate ab261702) or B2M knockout HEK-293T cell line Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line ab266828 (knockout cell lysate Human B2M (beta 2 Microglobulin) knockout HEK-293T cell lysate ab256845). To generate this image, wild-type and B2M knockout A431 and HEK-293T cell lysates were analysed.
    Nitrocellulose membranes were blocked in fluorescent western blot (TBS-based) blocking solution in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed in TBS-T, incubated with secondary antibodies Goat anti-Rabbit 800CW and Goat anti-Mouse 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.

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

    Lane 1: Wild-type HEK-293T cell lysate at 20 µg

    Lane 2: B2M knockout HEK-293T cell lysate at 20 µg

    Lane 2: Western blot - Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line (Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line ab266828)

    Lane 2: Western blot - Human B2M (beta 2 Microglobulin) knockout HEK-293T cell lysate (Human B2M (beta 2 Microglobulin) knockout HEK-293T cell lysate ab256845)

    Lane 3: Wild-type A431 cell lysate at 20 µg

    Lane 4: B2M knockout A431 cell lysate at 20 µg

    Lane 4: Western blot - Human B2M (beta 2 Microglobulin) knockout A-431 cell line (Human B2M (beta 2 Microglobulin) knockout A-431 cell line ab261893)

    Lane 4: Western blot - Human B2M (beta 2 Microglobulin) knockout A-431 cell lysate (Human B2M (beta 2 Microglobulin) knockout A-431 cell lysate ab261702)

    Performed under reducing conditions.

    Predicted band size: 14 kDa

    Observed band size: 12 kDa

  • Immunoprecipitation - Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853), expandable thumbnail

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

    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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab75853 in Raji whole cell lysate.

    For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10,000 dilution.

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    All lanes: Immunoprecipitation - Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853)

    Predicted band size: 14 kDa

    Observed band size: 14 kDa

  • Flow Cytometry (Intracellular) - Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853), expandable thumbnail

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

    Intracellular 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.

  • Western blot - Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853), expandable thumbnail

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

    Blocking and dilution buffer: 5% NFDM /TBST.

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

    Lane 1: U937 whole cell lysate at 20 µg

    Lane 2: HeLa whole cell lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 14 kDa

    Observed band size: 14 kDa

  • Immunocytochemistry/ Immunofluorescence - Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853), expandable thumbnail

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

    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. Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) 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. Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291, a mouse anti-tubulin (1/1000) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).

    Control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (1/1000) and secondary antibody, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).

  • Western blot - Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853), expandable thumbnail

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

    Blocking and dilution buffer: 5% NFDM /TBST.

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

    Lane 1: Mouse kidney tissue lysate at 20 µg

    Lane 2: Rat kidney tissue lysate at 20 µg

    Secondary

    All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution

    Predicted band size: 14 kDa

    Observed band size: 14 kDa

  • Western blot - Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853), expandable thumbnail

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

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

    Lane 1: Raji cell lysate at 10 µg

    Lane 2: U937 cell lysate at 10 µg

    Lane 3: HeLa cell lysate at 10 µg

    Lane 4: Human bone marrow lysate at 10 µg

    Secondary

    All lanes: HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

    Predicted band size: 14 kDa

    Observed band size: 14 kDa

  • Western blot - Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853), expandable thumbnail

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

    Western blot: Anti-B2M antibody [EP2978Y] (ab75853) staining at 1/5000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab75853 was shown to bind specifically to B2M. A band was observed at 13 kDa in wild-type A549 cell lysates with no signal observed at this size in B2M knockout cell line. To generate this image, wild-type and B2M knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

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

    Lane 1: Wild-type A549 cell lysate at 20 µg

    Lane 2: B2M knockout A549 cell lysate at 20 µg

    Lane 3: Wild-type HEK-293T ab255553 cell lysate at 20 µg

    Lane 4: B2M knockout HEK-293T Human B2M (beta 2 Microglobulin) knockout HEK-293T cell line ab266828 cell lysate at 20 µg

    Secondary

    All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution

    Performed under reducing conditions.

  • Flow Cytometry (Intracellular) - Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853), expandable thumbnail

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

    Flow cytometry overlay histogram showing C57 BL/6 mouse splenocytes stained with ab75853 (red line). Cells were fixed and permeabilised with BD Cytofix/Cytoperm™ for 20 min. The cells were incubated in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody (ab75853) (1x 106 in 100μl at 0.2 μg/ml (1/11300)) for 30min on ice.

    The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice

    Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

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Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

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