Phospho-SMC1 (S957) Monoclonal Antibody

• Catalog Number: orb548532
• Category: Antibodies
• Description: Phospho-SMC1 (S957) Monoclonal Antibody
• Species/Host: Rabbit
• Clonality: Monoclonal
• Clone Number: FCI-19
• Tested applications: ICC, IF, IHC, WB
• Reactivity: Human
• Isotype: Rabbit IgG
• Immunogen: A synthesized peptide derived from human Phospho-SMC1 (S957) Structural maintenance of chromosomes 1 (SMC1) protein is a chromosomal protein member of the cohesin complex that enables sister chromatid cohesion and plays a role in DNA repair. ATM/NBS1-dependent phosphorylation of SMC1 occurs at Ser957 and Ser966 in response to ionizing radiation (IR) as part of the intra-S-phase DNA damage checkpoint. SMC1 phosphorylation is ATM-independent in cells subjected to other forms of DNA damage, including UV light and hydroxyurea treatment.
• Concentration: Actual concentration vary by lot. Use suggested dilution ratio to decide dilution procedure.
• Form/Appearance: Liquid
• Conjugation: Unconjugated
• UniProt ID: Q14683
• 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: WB 1:500-1:2000IHC 1:50-1:200ICC/IF 1:50-1:200
• Expiration Date: 12 months from date of receipt.

Western blot analysis of SMC1A expression in (1) HeLa cell lysate; (2) HeLa cells lysate treated with AP using anti-SMC1A antibody (orb548532). 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 50ug of sample under reducing conditions. 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 rabbit anti-SMC1A antigen affinity purified polyclonal antibody (Catalog # orb548532) at 0.5 ug/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 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 (Catalog # orb90444) with Tanon 5200 system. A specific band was detected for SMC1A.

IHC analysis of SMC1A using anti-SMC1A antibody (orb548532). SMC1A was detected in paraffin-embedded section. 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 1ug/ml rabbit anti-SMC1A Antibody (orb548532) overnight at 4°C. Biotinylated goat anti Rabbit IgG IgG antibody was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using Strepavidin-Biotin-Complex (SABC) (Catalog # orb90444) with DAB as the chromogen.