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    beta-2-microglobulin-antibody-epr21752-214-ab218230.pdf

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Cardiovascular Blood Serum Proteins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Western blot - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
  • Western blot - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
  • Flow Cytometry (Intracellular) - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
  • Western blot - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
  • Immunoprecipitation - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
  • Flow Cytometry (Intracellular) - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
  • 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

Conjugates logo Related conjugates and formulations

Carrier Free

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Anti-Sonic Hedgehog antibody [EP1190Y] - BSA and Azide free (ab175180)
Knockout
Product image
Human B2M (beta 2 Microglobulin) knockout Hep G2 cell line (ab262325)

View more associated products

Overview

  • 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.

  • 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.
  • 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
    For more information see here.

    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

  • 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...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR21752-214
  • Isotype

    IgG
  • Research areas

    • Cardiovascular
    • Blood
    • Serum Proteins
    • Immunology
    • Adaptive Immunity
    • MHC
    • Class I
    • Cancer
    • Tumor biomarkers
    • Globulins
    • Cardiovascular
    • Blood
    • Blood Cell Antigens
    • WBC Antigens
    • HLA
    • Kits/ Lysates/ Other
    • Kits
    • ELISA Kits
    • ELISA Kits
    • Blood cell antigens ELISA kits

Associated products

  • Alternative Versions

    • Anti-beta 2 Microglobulin antibody [EPR21752-214] - BSA and Azide free (ab237032)
  • Compatible Secondaries

    • VeriBlot for IP Detection Reagent (HRP) (ab131366)
    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human B2M (beta 2 Microglobulin) knockout Hep G2 cell line (ab262325)
  • KO cell lysates

    • Human B2M (beta 2 Microglobulin) knockout Hep G2 cell lysate (ab256846)
  • KO cell pellets

    • Human B2M (beta 2 Microglobulin) knockout Hep G2 cell pellet (ab278904)
  • Recombinant Protein

    • Recombinant Human beta 2 Microglobulin protein (ab98116)
  • Related Products

    • Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

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
WB
1/1000. Detects a band of approximately 12 kDa (predicted molecular weight: 14 kDa).
IHC-P (2)
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.
Notes
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

  • 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.
  • Target information above from: UniProt accession P61769 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 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
    • Unigene: 702947 Human
    see all
  • 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

Images

  • Western blot - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
    Western blot - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
    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.

  • Western blot - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
    Western blot - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
    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 kDa



    Lanes 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)

    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.

  • Flow Cytometry (Intracellular) - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
    Flow Cytometry (Intracellular) - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)

    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. 

     

     

  • Western blot - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
    Western blot - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
    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 kDa



    Blocking/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.

  • Immunoprecipitation - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
    Immunoprecipitation - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)

    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)

    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.

  • Flow Cytometry (Intracellular) - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
    Flow Cytometry (Intracellular) - Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)

    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. 

     

     

  • Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)
    Anti-beta 2 Microglobulin antibody [EPR21752-214] (ab218230)

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.

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (1)

Publishing research using ab218230? Please let us know so that we can cite the reference in this datasheet.

ab218230 has been referenced in 1 publication.

  • 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

Customer reviews and Q&As

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1-2 of 2 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-beta 2 Microglobulin antibody [EPR21752-214]

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
Permeabilization
No
Specification
kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: RT°C
Fixative
10% NBF
Read More

Abcam user community

Verified customer

Submitted May 16 2019

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-beta 2 Microglobulin antibody [EPR21752-214]

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (kidney)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA pH 9.0
Permeabilization
No
Specification
kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 15% · Temperature: RT°C
Fixative
10% NBF
Read More

Abcam user community

Verified customer

Submitted May 16 2019

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