Anti-Topoisomerase I antibody

• Supplier: Abcam
• Catalog: ab85038
• Prices: $385.00
• Sizes: 100 µg
• Host: Rabbit
• Clonality: Polyclonal
• Isotype: IgG
• Applications: ICC/IF ICC/IF WB IHC-P IP
• Species Reactivities: Mouse, Rat, Human, Chicken, Hamster, Bovine, Monkey
• Antigen: Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human Topoisomerase I
• Description: Rabbit Polyclonal
• Gene: TOP1
• Conjugate: Unconjugated

Product images

All lanes : Anti-Topoisomerase I antibody (ab85038) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell LysateLane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell LysateLane 4 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 6 : Caco 2 (Human colonic carcinoma cell line) Whole Cell LysateLane 7 : SHSY-5Y (Human neuroblastoma cell line) Whole Cell LysateLysates/proteins at 10 µg per lane.SecondaryGoat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

All lanes : Anti-Topoisomerase I antibody (ab85038) at 1 µg/mlLane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate Lane 2 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate Lysates/proteins at 10 µg per lane.SecondaryGoat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilutiondeveloped using the ECL techniquePerformed under reducing conditions.

Topoisomerase I was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Rabbit polyclonal to Topoisomerase I and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Hek293 whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab85038.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 90kDa; Topoisomerase I

ICC/IF image of ab85038 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab85038, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 4% PFA fixed (10 min) Hek293, HepG2 and MCF7 cells at 1µg/ml, and in 100% methnaol fixed (5 min) HeLa cells at 1µg/ml.

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