![GPR2/CCR10 was immunoprecipitated using 0.5mg MCF7 whole cell extract, 5µg of Rabbit polyclonal to GPR2/CCR10 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, MCF7 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 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab30718.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 42kDa; GPR2/CCR10](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_2/27206_ab30718-171837-IPV019ab3071820mMod.jpg)
GPR2/CCR10 was immunoprecipitated using 0.5mg MCF7 whole cell extract, 5µg of Rabbit polyclonal to GPR2/CCR10 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, MCF7 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 70°C; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab30718.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 42kDa; GPR2/CCR10
Anti-GPR2/CCR10 antibody (ab30718) at 1/250 dilution + MCF-7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µgSecondaryIRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/15000 dilutionPerformed under reducing conditions.
Image courtesy of Human Protein Atlasab30718 staining GPCR GPR2/CCR10 in Human smooth muscle. The praffin embedded tissue was incubated with ab30718 (1/2000 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. Ab30718 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
ICC/IF image of ab30718 stained human HEK 293 cells. The cells were methanol fixed (5 min), permabilised in 0.1% PBS-Tween (20 min) and incubated with the antibody (ab30718, 5µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to 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).