Recombinant Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR21752-214] to beta 2 Microglobulin
- Suitable for: WB, IHC-P, IP, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-beta 2 Microglobulin antibody [EPR21752-214]
See all beta 2 Microglobulin primary antibodies -
Description
Rabbit monoclonal [EPR21752-214] to beta 2 Microglobulin -
Host species
Rabbit -
Tested applications
Suitable for: WB, IHC-P, IP, Flow Cyt (Intra)more details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Full length native protein (purified). This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: HeLa, Jurkat and HepG2 cell lysate; Mouse serum and plasma; Rat serum and plasma; Human plasma; Human skin, kidney and liver lysate, Wild-type HEK-293T cell lysate. IHC-P: Human spleen and kidney tissue. Flow Cyt (intra): HeLa and HepG2 cells. IP: HeLa cell lysate.
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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 long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR21752-214 -
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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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab218230 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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WB |
1/1000. Detects a band of approximately 12 kDa (predicted molecular weight: 14 kDa).
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IHC-P | (2) |
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IP |
1/30.
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Flow Cyt (Intra) |
1/50.
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Notes |
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WB
1/1000. Detects a band of approximately 12 kDa (predicted molecular weight: 14 kDa). |
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
IP
1/30. |
Flow Cyt (Intra)
1/50. |
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 [EPR21752-214] (ab218230) at 1/1000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : B2M knockout HEK-293T 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 [EPR21752-214] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab218230 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 [EPR21752-214] (ab218230) at 1/500 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 - ab218230 observed at 14 kDa. Red - loading control ab8245 observed at 36 kDa.
ab218230 Anti-beta 2 Microglobulin antibody [EPR21752-214] 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. ab218230 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 500 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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Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling beta 2 Microglobulin with ab218230 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody. Positive staining on endothelial cells of human kidney is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HepG2 (human liver hepatocellular carcinoma cell line) cell line labeling beta 2 Microglobulinwith ab218230 at 1/500 dilution (red) compared with a Rabbit monoclonal IgG - Isotype control (ab172730) (black) and an unlabeled control cells incubatedwith secondary antibody only) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
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All lanes : Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230) at 1/1000 dilution
Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 2 : Jurkat (human T cell leukemia T lymphocyte), whole cell lysate
Lane 3 : HepG2 (human liver hepatocellular carcinoma cell line) cell lysate
Lane 4 : Mouse serum
Lane 5 : Mouse plasma
Lane 6 : Rat serum
Lane 7 : Rat plasma
Lane 8 : Human plasma
Lane 9 : Human skin lysate
Lane 10 : Human kidney lysate
Lane 11 : Human liver lysate
Lysates/proteins at 10 µg per lane.
Secondary
Lanes 1-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Lanes 4-11 : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/1000 dilution
Predicted band size: 14 kDaBlocking/diluting buffer and concentration: 5% NFDM/TBST.
Exposure times: Lane 1: 6 seconds; Lane 2-3: 37 seconds: Lane 4-7: 48 seconds; Lane 8-11: 3 minutes.
Lanes 4-11: This blot was developed using a higher sensitive ECL substrate.
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Beta 2 Microglobulin was immunoprecipitated from 0.35 mg HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab218230 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab218230 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.
Lane 1: HeLa whole cell lysate 10 µg (Input).
Lane 2: ab218230 IP in HeLa whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab218230 in HeLa whole cell lysate (-).
Blocking/Dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 minutes. -
Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling beta 2 Microglobulin with ab218230 at 1/4000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody. Positive staining on endothelial cells of human spleen is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cell line labeling beta 2 Microglobulinwith ab218230 at 1/50 dilution (red) compared with a Rabbit monoclonal IgG - Isotype control (ab172730) (black) and an unlabeled control (cells incubatedwith secondary antibody only) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (4)
ab218230 has been referenced in 4 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
- Ren J et al. Histone methyltransferase WHSC1 loss dampens MHC-I antigen presentation pathway to impair IFN-γ-stimulated antitumor immunity. J Clin Invest 132:N/A (2022). PubMed: 35230972
- Fu Y et al. An HNSCC syngeneic mouse model for tumor immunology research and preclinical evaluation. Int J Mol Med 46:1501-1513 (2020). PubMed: 32700748
- Liu Y et al. A novel rat model of ocular hypertension by a single intracameral injection of cross-linked hyaluronic acid hydrogel (Healaflow® ). Basic Clin Pharmacol Toxicol 127:361-370 (2020). PubMed: 32383327