MA1-19141

antibody from Invitrogen Antibodies

Targeting: B2M

Western blot (Supportive data in Antibodypedia)

Antibody Data

Product number

MA1-19141

Provider

Invitrogen Antibodies

Product name

beta-2 Microglobulin Monoclonal Antibody (B2M-01)

Antibody type

Monoclonal

Antigen

Purifed from natural sources

Description

This antibody reacts with beta2-microglobulin associated with cell-surface MHC Class I molecules and other membrane antigens as well as with soluble beta2-microglobulin. It will not cross-react with mouse, rabbit, canine, chicken or bovine.

Antibody clone number

B2M-01

Concentration

1 mg/mL

References

Submitted references

Presentation and binding affinity of equine infectious anemia virus CTL envelope and matrix protein epitopes by an expressed equine classical MHC class I molecule.

McGuire TC, Leib SR, Mealey RH, Fraser DG, Prieur DJ
Journal of immunology (Baltimore, Md. : 1950) 2003 Aug 15;171(4):1984-93

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of beta-2 Microglobulin using a monoclonal antibody (Product # MA1-19141).

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blotting analysis (non-reducing conditions) of whole cell lysate of various cell lines using anti-human β2-microglobulin (B2M-01) Monoclonal antibody (Product # MA1-19141). Lane 1: RAJI human Burkitt lymphoma cell line; Lane 2: EL4 mouse lymphoblastic lymphoma cell line; Lane 3: U937 human histiocytic lymphoma cell line.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockout of B2M was achieved by CRISPR-Cas9 genome editing using LentiArray™ Lentiviral sgRNA (Product # A32042, Assay ID CRISPR983707_LV) and LentiArray Cas9 Lentivirus (Product # A32064). Western blot analysis of B2M was performed by loading 50 µg of HeLa Wild Type (Lane 1), HeLa Cas9 (Lane 2) andHeLa B2M KO (Lane 3) membrane enriched extracts. The samples were electrophoresed using Novex™ 16%, Tricine, 1.0 mm, Mini Protein Gel (Product # EC66952BOX). Resolved proteins were then transferred onto a nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with Anti-beta-2 Microglobulin Monoclonal Antibody (B2M-01) (Product # MA1-19141, 5 µg/mL dilution) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:5,000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Atto Ultimate Sensitivity Substrate (Product # A38556). Loss of signal upon CRISPR mediated knockout (KO) using the LentiArray™ CRISPR product line confirms that antibody is specific to B2M.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Knockdown of beta-2 Microglobulin was achieved by transfecting HeLa with beta-2 Microglobulin specific siRNAs (Silencer® select Product # s1854, s1853). Western blot analysis (Fig. a) was performed using Whole cell extracts from the beta-2 Microglobulin knockdown cells (lane 3), non-targeting scrambled siRNA transfected cells (lane 2) and untransfected cells (lane 1). The blot was probed with beta-2 Microglobulin Monoclonal Antibody (B2M-01) (Product # MA1-19141, 3 microgram/ml) and Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution). Densitometric analysis of this western blot is shown in histogram (Fig. b). Decrease in signal upon siRNA mediated knock down confirms that antibody is specific to beta-2 Microglobulin.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot was performed using Anti-beta-2 Microglobulin Monoclonal Antibody (B2M-01) (Product # MA1-19141) and a 12 kDa band corresponding to beta-2 Microglobulin was observed across all cell lines tested except for SH-SY5Y and NTERA2 cl.D1, which are reported to express low levels of beta-2 Microglobulin. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), THP-1 (Lane 2), U-937 (Lane 3), SH-SY5Y (Lane 4) and NTERA-2 cl.D1 (Lane 5) were electrophoresed using Novex™ 16% Tricine Protein Gel (Product # EC6695BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23001) by iBlot® 2 Dry Blotting System (Product # IB21001). The blot was probed with the primary antibody (3 µg/mL) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177,1:4000 dilution) using the iBright FL1000 (Product # A32752). Chemiluminescent detection was performed using SuperSignal™ West Dura Extended Duration Substrate (Product # 34076).