The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration.
Use a concentration of 1.25 µg/ml. Detects a band of approximately 57 kDa (predicted molecular weight: 57 kDa).Can be blocked with Human RIP3 peptide (ab178834). Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
Use a concentration of 3 - 12 µg/ml.
Detected at lower levels in heart, placenta, lung and kidney. Isoform 3 is significantly increased in colon and lung cancers.
Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. Contains 1 protein kinase domain.
Immunohistochemical analysis of Formalin/PFA-fixed paraffin-embedded human pancreas sections labelling RIP3 with ab56164 at 3 - 12 µg/mL. The secondary antibody used was a donkey anti-rabbit antibody at 1/200. The exposure time was 0.2 seconds at magnification of 20X.
RIP3 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to RIP3 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, Jurkat 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 ab56164. Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697). Band: 65kDa; RIP3.
Western blot - RIP3 antibody (ab56164)
All lanes : Anti-RIP3 antibody (ab56164) at 1.25 µg/ml
Lane 1 : Molecular weight markers Lane 2 : Jurkat cell lysate at 10 µg
Secondary HRP conjugated anti-Rabbit IgG at 1/50000 dilution
Predicted band size : 57 kDa Observed band size : 57 kDa Gel concentration: 12%