Anti-Beta 2 Microglobulin/B2M Antibody Picoband®

Figure 1. Western blot analysis of Beta 2 Microglobulin using anti-Beta 2 Microglobulin antibody (A00456-1).
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 (Catalog # A00456-1) 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 (Catalog # EK1002) 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.

Figure 2. IHC analysis of Beta 2 Microglobulin using anti- Beta 2 Microglobulin antibody (A00456-1).
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 1μg/ml rabbit anti- Beta 2 Microglobulin Antibody (A00456-1) 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)(Catalog # SA1022) with DAB as the chromogen.

Figure 3. IHC analysis of Beta 2 Microglobulin using anti- Beta 2 Microglobulin antibody (A00456-1).
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 1μg/ml rabbit anti- Beta 2 Microglobulin Antibody (A00456-1) 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)(Catalog # SA1022) with DAB as the chromogen.

Figure 4. IF analysis of Beta 2 Microglobulin using anti- Beta 2 Microglobulin antibody (A00456-1).
Beta 2 Microglobulin was detected in immunocytochemical section of A431 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti- Beta 2 Microglobulin Antibody (A00456-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.

Figure 5. IF analysis of Beta 2 Microglobulin using anti- Beta 2 Microglobulin antibody (A00456-1).
Beta 2 Microglobulin was detected in immunocytochemical section of A431 cell. Enzyme antigen retrieval was performed using IHC enzyme antigen retrieval reagent (AR0022) for 15 mins. The cells were blocked with 10% goat serum. And then incubated with 2μg/mL rabbit anti- Beta 2 Microglobulin Antibody (A00456-1) overnight at 4°C. DyLight®488 Conjugated Goat Anti-Rabbit IgG (BA1127) 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.

Figure 6. Flow Cytometry analysis of U251 cells using anti- Beta 2 Microglobulin antibody (A00456-1).
Overlay histogram showing U251 cells stained with A00456-1 (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 (A00456-1,1μg/1x10⁶ cells) for 30 min at 20°C. DyLight488 conjugated goat anti-rabbit IgG (BA1127, 5-10μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1μg/1x10⁶) used under the same conditions. Unlabelled sample without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Figure 7. 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) (EZ1691).