Anti-Beta 2 Microglobulin B2M Antibody (monoclonal, 2H10)

• Catalog Number: orb507558
• Category: Antibodies
• Description: Anti-Beta 2 Microglobulin B2M Antibody (monoclonal, 2H10). Tested in Flow Cytometry, IF, IHC, ICC, WB applications. This antibody reacts with Human, Monkey.
• Species/Host: Mouse
• Clonality: Monoclonal
• Clone Number: 2H10
• Tested applications: FC, ICC, IF, IHC, WB
• Reactivity: Human, Monkey
• Isotype: Mouse IgG2b
• Immunogen: E.coli-derived human Beta 2 Microglobulin recombinant protein (Position: Q22-M119). Human Beta 2 Microglobulin shares 69.4% and 74.5% amino acid (aa) sequence identity with mouse and rat Beta 2 Microglobulin, respectively.
• Antibody Type: Primary Antibody
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Form/Appearance: Lyophilized
• Conjugation: Unconjugated
• MW: 12 kDa
• UniProt ID: P61769
• Storage: Maintain refrigerated at 2-8°C for up to 2 weeks. For long term storage store at -20°C in small aliquots to prevent freeze-thaw cycles.
• Alternative names: Beta-2-microglobulin; Beta-2-microglobulin form pI
• Note: For research use only
• Application notes: Western blot, 0.1-0.5μg/ml Immunohistochemistry (Paraffin-embedded Section), 0.5-1μg/ml Immunocytochemistry/Immunofluorescence, 2μg/ml Flow Cytometry (Fixed), 1-3μg/1x106 cells. Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml
• Expiration Date: 12 months from date of receipt.

Flow Cytometry analysis of A431 cells using anti-Beta 2 Microglobulin antibody. Overlay histogram showing A431 cells (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with mouse anti-Beta 2 Microglobulin Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-mouse IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was mouse IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

IF analysis of Beta 2 Microglobulin using anti-Beta 2 Microglobulin antibody. Beta 2 Microglobulin was detected in immunocytochemical section of A431 cells. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2 µg/mL mouse anti-Beta 2 Microglobulin Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Mouse IgG was used as secondary antibody at 1:100 dilution and incubated for 30 minutes at 37°C. The section was counterstained with DAPI. Visualize using a fluorescence microscope and filter sets appropriate for the label used.

IHC analysis of Beta 2 Microglobulin using anti-Beta 2 Microglobulin antibody. Beta 2 Microglobulin was detected in paraffin-embedded section of human intestinal cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-Beta 2 Microglobulin Antibody overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

IHC analysis of Beta 2 Microglobulin using anti-Beta 2 Microglobulin antibody. Beta 2 Microglobulin was detected in paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-Beta 2 Microglobulin Antibody overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

IHC analysis of Beta 2 Microglobulin using anti-Beta 2 Microglobulin antibody. Beta 2 Microglobulin was detected in paraffin-embedded section of human mammary cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml mouse anti-Beta 2 Microglobulin Antibody overnight at 4°C. Biotinylated goat anti-mouse IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) with DAB as the chromogen.

Western blot analysis of Beta 2 Microglobulin using anti-Beta 2 Microglobulin antibody. Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 50 ug of sample under reducing conditions. Lane 1: COS-7 whole cell lysates, Lane 2: HELA whole cell lysates, Lane 3: HL-60 whole cell lysates, Lane 4: HEPG2 whole cell lysates, Lane 5: K562 whole cell lysates, Lane 6: 293T whole cell lysates. After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT. The membrane was incubated with mouse anti-Beta 2 Microglobulin antigen affinity purified monoclonal antibody at 0.5 µg/mL overnight at 4°C, then washed with TBS-0.1% Tween 3 times with 5 minutes each and probed with a goat anti-mouse IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system. A specific band was detected for Beta 2 Microglobulin at approximately 12KD. The expected band size for Beta 2 Microglobulin is at 12KD.