Anti-Calpain 1 Antibody (monoclonal, 2E3)

• Catalog Number: orb763101
• Category: Antibodies
• Description: Anti-Calpain 1 Antibody (monoclonal, 2E3). Tested in Flow Cytometry, WB applications. This antibody reacts with Human.
• Clonality: Monoclonal
• Species/Host: Mouse
• Isotype: Mouse IgG2b
• Conjugation: Unconjugated
• Reactivity: Human
• Form/Appearance: Lyophilized
• Concentration: Adding 0.2 ml of distilled water will yield a concentration of 500 μg/ml.
• Immunogen: E.coli-derived human Calpain 1 recombinant protein (Position: Q396-A555). Human Calpain 1 shares 86% amino acid (aa) sequence identity with both mouse and rat Calpain 1.
• UniProt ID: P07384
• MW: 82 kDa
• Tested applications: FC, WB
• Application notes: Western blot, 0.25-0.5μg/ml, Human Flow Cytometry (Fixed), 1-3μg/1x106 cells, Human. Add 0.2ml of distilled water will yield a concentration of 500ug/ml
• Antibody Type: Primary Antibody
• Clone Number: 2E3
• 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: Structural maintenance of chromosomes protein 3; S
• Note: For research use only

Flow Cytometry analysis of A549 cells using anti-Calpain 1 antibody. Overlay histogram showing A549 cells (Blue line). The cells were fixed with 4% paraformaldehyde and blocked with 10% normal goat serum. And then incubated with mouse anti-Calpain 1 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 without incubation with primary antibody and secondary antibody (Red line) was used as a blank control.

Western blot analysis of Calpain 1 using anti-Calpain 1 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 Jurkat whole cell lysates, Lane 2: human Hela whole cell lysates, Lane 3: human K562 whole cell lysates, Lane 4: human PC-3 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 mouse anti-Calpain 1 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 1:10000 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 Calpain 1 at approximately 82 KD. The expected band size for Calpain 1 is at 82 KD.