Anti-Beta 2 Microglobulin/B2M Antibody

• Catalog Number: orb18580
• Category: Antibodies
• Description: Anti-Beta 2 Microglobulin/B2M Antibody
• Species/Host: Rabbit
• Clonality: Polyclonal
• Tested applications: ELISA, FC, ICC, IF, IHC, WB
• Reactivity: Human
• Isotype: Rabbit IgG
• 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: 14 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.
• Note: For research use only
• Application notes: Western blot, 0.1-0.5μg/ml, Human Immunohistochemistry(Paraffin-embedded Section), 0.5-1 μg/ml, Human Immunocytochemistry/Immunofluorescence, 2μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human ELISA(Cap), 1-5 μg/ml, Human. Adding 0.2ml of distilled water will yield a concentration of 500ug/ml
• Expiration Date: 12 months from date of receipt.

Flow Cytometry analysis of U251 cells using anti-Beta 2 Microglobulin antibody. Overlay histogram showing U251 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 rabbit anti-Beta 2 Microglobulin Antibody (1 µg/1x10^6 cells) for 30 min at 20°C. DyLight488 conjugated goat anti-rabbit IgG (5-10 µg/1x10^6 cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 µg/1x10^6) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

IF analysis of Beta 2 Microglobulin using anti-Beta 2 Microglobulin antibody. Beta 2 Microglobulin was detected in immunocytochemical section of A431 cell. 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 rabbit anti-Beta 2 Microglobulin Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit 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.

IF analysis of Beta 2 Microglobulin using anti-Beta 2 Microglobulin antibody. Beta 2 Microglobulin was detected in immunocytochemical section of A431 cell. 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 rabbit anti-Beta 2 Microglobulin Antibody overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit 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 tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-Beta 2 Microglobulin Antibody overnight at 4°C. Biotinylated goat anti-rabbit 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 tissues. Heat mediated antigen retrieval was performed in citrate buffer (pH6, epitope retrieval solution) for 20 mins. The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 1 µg/ml rabbit anti-Beta 2 Microglobulin Antibody overnight at 4°C. Biotinylated goat anti-rabbit 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.

Sandwich ELISA - Recombinant human Beta 2 Microglobulin/B2M protein standard curve. Use in combination with reagents from Human Beta 2 Microglobulin/B2M ELISA Kit EZ-Set (DIY Antibody Pairs).

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 30 ug of sample under reducing conditions. Lane 1: human U251 whole cell lysates, Lane 2: human THP-1 whole cell lysates, Lane 3: human Hela whole cell lysates, Lane 4: human A431 whole cell lysates. After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Beta 2 Microglobulin antigen affinity purified polyclonal 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-rabbit 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 14 kDa. The expected band size for Beta 2 Microglobulin is at 14 kDa.