Anti-CSL (CBF1, RBP-Jk, J kappa-Recombination Signal-Binding Protein, Suppressor of Hairless, Lag-1)

CSL (CBF1/RBP-Jk, Su(H)/Suppressor of Hairless, Lag-1) is a 56kD member of the Su(H) family of transcription factors. It is constitutively bound to DNA where it participates in gene repression. Upon ligand binding, the intracellular domain of Notch translocates to the nucleus and interacts with CSL, allowing transcription of Notch target genes. Human CSL is 500 amino acids (aa) in length. It contains three DNA binding sites (aa # 58-65, # 192-201, and # 265-297) and one IPT domain (aa # 355-445). Multiple splice variants exist that involve the first 89 N-terminal amino acids. One variant (isoform 2) is 487 aa in length and shows a 6 aa substitution for amino acids # 1-19. Over amino acids # 195-487, human CSL isoform 2 shares more than 98% aa sequence identity with mouse, rat, and canine CSL. Applications: Western Blot: This antibody can be used at 0.1-0.2ug/ml with the appropriate secondary reagents to detect human CSL. The detection limit for rhCSL is approximately 10 ng/lane under non-reducing and reducing conditions. , Immunocytochemistry: This antibody can be used at a concentration of 10ug/ml to detect human CSL on HASMC (human aortic smooth muscle) cells. Cells were fixed with PBS containing 4% paraformaldehyde for 20 minutes at room temperature and blocked with PBS containing 10% normal donkey serum, 0.1% Triton X-100, and 1% BSA for 45 minutes at room temperature. After blocking, cells were incubated with diluted primary antibody overnight at 4° C followed by NorthernLightsTM 557-coupled donkey anti-goat IgG at room temperature in the dark for one hour. Between each step, cells were washed with PBS containing 0.1% BSA. , Direct ELISA: This antibody can be used at 0.5-1.0ug/ml with the appropriate secondary reagents to detect human CSL. The detection limit for rhCSL is approximately 5 ng/well. , Optimal dilutions should be determined by each laboratory for each application. Storage and Stability: Lyophilized powder may be stored at 4°C for short-term only. Reconstitute to nominal volume by adding sterile PBS with 40-50% glycerol and store at -20°C. Reconstituted product is stable for 12 months at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.