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.
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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.
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.
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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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
Performed under reducing conditions.
Predicted band size: 14 kDa
Observed band size: 12 kDa why is the actual band size different from the predicted?False colour image of 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 HEK-293T cell lysates with no signal observed at this size in B2M knockout cell line ab266828 (knockout cell lysate ab256845). To generate this image, wild-type and B2M knockout HEK-293T 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 (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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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 (33)
ab75853 has been referenced in 33 publications.
- Xu P et al. Inhibition of innate immune response ameliorates Zika virus-induced neurogenesis deficit in human neural stem cells. PLoS Negl Trop Dis 15:e0009183 (2021). PubMed: 33657175
- Montenont E et al. CRISPR-edited megakaryocytes for rapid screening of platelet gene functions. Blood Adv 5:2362-2374 (2021). PubMed: 33944898
- Martinotti S et al. Manuka Honey Induces Apoptosis of Epithelial Cancer Cells through Aquaporin-3 and Calcium Signaling. Life (Basel) 10:N/A (2020). PubMed: 33120979
- Brenner E et al. Cancer immune control needs senescence induction by interferon-dependent cell cycle regulator pathways in tumours. Nat Commun 11:1335 (2020). PubMed: 32165639
- Fernandes E et al. Nucleolin-Sle A Glycoforms as E-Selectin Ligands and Potentially Targetable Biomarkers at the Cell Surface of Gastric Cancer Cells. Cancers (Basel) 12:N/A (2020). PubMed: 32252346