Anti GST

Immunoprecipitation of anti GST antibody. Lane 1: T98G cells incubated with GST Pin1. Lane 2: T98G cells incubated with GST CDK4/cyclinD1. Lane 3: T98G cells incubated with GST Pin1 and GST CDK4/cyclinD1. Immunoprecipitated with pRb antibody. Load: 25 ug per lane. Primary antibody: anti GST 1:400 for overnight at 4C. Secondary antibody: IRDye800TM secondary antibody at 1:10,000 for 45 min at RT. Block: 5% BLOTTO overnight at 4C.

ELISA results of purified Goat Anti-GST Antibody tested against GST. Each well was coated in 1.0 ug of antigen. The starting dilution of antibody was 5 ug/mL and the X-axis represents the Log10 of a 3-fold dilution. This titration is a 4-parameter curve fit where the IC50 is defined as the titer of the antibody. Assay performed using Buffer (p/n MB-060-1000), Substrate (p/n TMB-8000), and Donkey Anti-Goat IgG Antibody HRP (p/n 605-703-125) at 1:15,000.

Western Blot of Goat Anti-GST Antibody. Lane 1: Opal Pre-stained Molecular Weight Marker (p/n MB-210-0500). Lane 2: rec GST [0.1ug]/ HeLa Whole Cell Lysate [10ug]. Lane 3: rec GST [0.05ug]/ HeLa Whole Cell Lysate [10ug]. Lane 4: rec GST [0.02ug]/ HeLa Whole Cell Lysate [10ug]. Lane 3: HeLa Whole Cell Lysate (p/n W09-000-364) [10ug]. Primary Antibody: Goat anti-GST antibody (p/n 600-101-200) at 1.0ug/mL for overnight at 2-8C. Secondary Antibody: Rabbit Anti-Goat IgG (H&L) Antibody Peroxidase Conjugated (p/n 605-4302) at 1:40,000 for 30 min at RT. Block: (p/n MB-070) at RT for 30 min. Predicted/Observed size: ~26kDa.

Western Blot of Goat anti-GST antibody. Marker: Opal Pre-stained ladder (p/n MB-210-0500). Lane 1: HEK293 lysate (p/n W09-000-365). Lane 2: HeLa Lysate (p/n W09-000-364). Lane 3: CHO/K1 Lysate (p/n W07-000-357). Lane 4: MDA-MB-231 (p/n W09-001-GK6). Lane 5: A431 Lysate (p/n W09-000-361). Lane 6: Jurkat Lysate (p/n W09-001-370). Lane 7: NIH/3T3 Lysate (p/n W10-000-358). Lane 8: E-coli HCP Control (p/n 000-001-J08). Lane 9: FLAG Positive Control Lysate (p/n W00-001-383). Lane 10: Red Fluorescent Protein (p/n 000-001-379). Lane 11: Green Fluorescent Protein (p/n 000-001-215). Lane 12: Glutathione-S-Transferase Protein (p/n 000-001-200). Lane 13: Maltose Binding Protein (p/n 000-001-385). Load: 10 ug of lysate or 50ng of purified protein per lane. Primary antibody: GST antibody at 1ug/mL overnight at 4C. Secondary antibody: Peroxidase goat secondary antibody (p/n 605-4302) at 1:30,000 for 60 min at RT. Blocking Buffer: 1% Casein-TTBS (p/n MB-082) for 30 min at RT. Predicted/Observed size: 26 kDa for GST.