All lanes : Anti-PACT (PKR activating protein) / PRKRA antibody (ab31967) at 1 µg/mlLane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell LysateLane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate with Human PACT (PKR activating protein) / PRKRA peptide (ab30768) at 1 µg/mlLysates/proteins at 10 µg per lane.SecondaryIRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilutionPerformed under reducing conditions.
ICC/IF image of ab31967 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in TBS-T (20 min) and incubated with the antibody (ab31967, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. 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). DAPI was used to stain the cell nuclei (blue).
All lanes : Anti-PACT (PKR activating protein) / PRKRA antibody (ab31967) at 1 µg/mlLane 1 : Testis (Mouse) Tissue Lysate - normal tissue Lane 2 : PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate Lysates/proteins at 10 µg per lane.SecondaryIRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilutionPerformed under reducing conditions.
![PACT (PKR activating protein) / PRKRA was immunoprecipitated using 0.5mg Mouse Testis tissue, 5µg of Rabbit polyclonal to PACT (PKR activating protein) / PRKRA 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, Mouse Testis tissue lysate 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 ab31967.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 34kDa; PACT (PKR activating protein) / PRKRA](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_4/6415_PACT-PKR-activating-protein-PRKRA-Primary-antibodies-ab31967-3.jpg)
PACT (PKR activating protein) / PRKRA was immunoprecipitated using 0.5mg Mouse Testis tissue, 5µg of Rabbit polyclonal to PACT (PKR activating protein) / PRKRA 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, Mouse Testis tissue lysate 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 ab31967.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 34kDa; PACT (PKR activating protein) / PRKRA