Recombinant Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP2978Y] to beta 2 Microglobulin
- Suitable for: Flow Cyt (Intra), ICC/IF, WB, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-beta 2 Microglobulin antibody [EP2978Y]
See all beta 2 Microglobulin primary antibodies -
Description
Rabbit monoclonal [EP2978Y] to beta 2 Microglobulin -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ICC/IF, WB, IPmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HepG2, Raji, U937, Jurkat and HeLa whole cell lysates and human bone marrow, rat and mouse kidney tissue lysates, Wild-type HEK-293T cell lysate, Wild-type A431 cell lysate ICC/IF: HeLa cells. IP: Raji cell lysate. Flow Cyt (intra): HeLa cells, C57 BL/6 mouse bone marrow cells.
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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
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
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP2978Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
- Hep G2 whole cell lysate (ab166833)
- Mouse kidney normal tissue lysate - total protein (ab29305)
- HeLa whole cell lysate (ab29545)
- Raji whole cell lysate (ab30124)
- Mouse kidney tissue lysate - total protein (0 days) (ab7261)
- Mouse kidney tissue lysate (14 days) (ab7262)
- Mouse kidney tissue lysate (7 days) (ab7263)
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Recombinant Protein
Applications
The Abpromise guarantee
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 |
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Flow Cyt (Intra) |
1/20 - 1/60.
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ICC/IF |
1/100 - 1/250.
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WB | (7) |
1/5000. Predicted molecular weight: 14 kDa.
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.
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Notes |
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Flow Cyt (Intra)
1/20 - 1/60. |
ICC/IF
1/100 - 1/250. |
WB
1/5000. Predicted molecular weight: 14 kDa. 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. |
Target
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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
modificationsGlycation of Ile-21 is observed in long-term hemodialysis patients. -
Cellular localization
Secreted. Detected in serum and urine. - Information by UniProt
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Database links
- Entrez Gene: 567 Human
- Entrez Gene: 12010 Mouse
- Entrez Gene: 24223 Rat
- Omim: 109700 Human
- SwissProt: P61769 Human
- SwissProt: P01887 Mouse
- SwissProt: P07151 Rat
- Unigene: 534255 Human
see all -
Alternative names
- B2M antibody
- B2MG_HUMAN antibody
- Beta 2 microglobin antibody
see all
Images
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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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All lanes : Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853) at 1/5000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : B2M knockout HEK-293T cell lysate
Lane 3 : Wild-type A431 cell lysate
Lane 4 : B2M knockout A431 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 14 kDa
Observed band size: 12 kDa why is the actual band size different from the predicted?Western blot: Anti-beta 2 Microglobulin antibody [EP2978Y] staining at 1/5000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (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 ab261893 (knockout cell lysate ab261702) or B2M knockout HEK-293T cell line ab266828 (knockout 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 : Anti-beta 2 Microglobulin antibody [EP2978Y] (ab75853) at 1/1000 dilution
Lane 1 : Wild-type HepG2 cell lysate
Lane 2 : B2M knockout HepG2 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : Jurkat 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: 14 kDa
Observed band size: 14 kDaLanes 1-4: Merged signal (red and green). Green - ab75853 observed at 14 kDa. Red - 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 ab262325 (knockout 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 (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.
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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.
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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).
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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
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 14 kDa
Observed band size: 14 kDaBlocking and dilution buffer: 5% NFDM /TBST.
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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
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 14 kDa
Observed band size: 14 kDaBlocking and dilution buffer: 5% NFDM /TBST.
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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, 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.
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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
All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Predicted band size: 14 kDa
Observed band size: 14 kDa
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (51)
ab75853 has been referenced in 51 publications.
- Gao Y et al. Inactivation of AMPK Leads to Attenuation of Antigen Presentation and Immune Evasion in Lung Adenocarcinoma. Clin Cancer Res 28:227-237 (2022). PubMed: 34667030
- Zhan L et al. LC3 and NLRC5 interaction inhibits NLRC5-mediated MHC class I antigen presentation pathway in endometrial cancer. Cancer Lett 529:37-52 (2022). PubMed: 34974132
- Xiong W et al. Smad4 Deficiency Promotes Pancreatic Cancer Immunogenicity by Activating the Cancer-Autonomous DNA-Sensing Signaling Axis. Adv Sci (Weinh) 9:e2103029 (2022). PubMed: 35064757
- Xiong X et al. Emerging enterococcus pore-forming toxins with MHC/HLA-I as receptors. Cell 185:1157-1171.e22 (2022). PubMed: 35259335
- Piirsalu M et al. Lipopolysaccharide-Induced Strain-Specific Differences in Neuroinflammation and MHC-I Pathway Regulation in the Brains of Bl6 and 129Sv Mice. Cells 11:N/A (2022). PubMed: 35326484