Would you like to visit your country specific website?Western blot analysis of extracts from various cell lines using RCAS1 (D2B6N) XPWestern blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human RCAS1 (hRCAS1-Myc/DDK; +), using RCAS1 (D2B6N) XPImmunoprecipitation of RCAS1 from HCT 116 extracts using Rabbit (DA1E) mAb IgG XPConfocal immunofluorescent analysis of MCF7 cells, untreated (left) or treated with Brefeldin A #9972 (5 μg/ml, 1 hr; right), using RCAS1 (D2B6N) XPFlow cytometric analysis of Jurkat cells using RCAS1 (D2B6N) XPWestern blot analysis of extracts from various cell lines using RCAS1 (D2B6N) XPWestern blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing Myc/DDK-tagged full-length human RCAS1 (hRCAS1-Myc/DDK; +), using RCAS1 (D2B6N) XPImmunoprecipitation of RCAS1 from HCT 116 extracts using Rabbit (DA1E) mAb IgG XPConfocal immunofluorescent analysis of MCF7 cells, untreated (left) or treated with Brefeldin A #9972 (5 μg/ml, 1 hr; right), using RCAS1 (D2B6N) XPFlow cytometric analysis of Jurkat cells using RCAS1 (D2B6N) XPSupplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Repeat washing step once more.Proceed to one of the following specific set of steps.Achieve higher quality immunofluorescent images using the efficient and cost-effective, pre-made reagents in our Aspirate liquid, then cover cells to a depth of 2–3 mm with 4% formaldehyde diluted in 1X PBS.Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly147 of human RCAS1 protein.Receptor binding cancer antigen expressed on SiSo cells (RCAS1) is also known as estrogen receptor-binding fragment-associated gene 9 (EBAG9). Indeed, it has been shown that RCAS1 overexpression negatively regulates the cytolytic function of CTLs by negatively regulating protein trafficking from the Explore pathways + proteins related to this product. During wound repair, blood monocytes are recruited into the wound bed where they differentiate into macrophages that secrete cytokines and regulate subsequent repair events. Proteins were extracted from plasma, isolated exosomes and supernatant samples using RIPA lysis buffer with protease inhibitor cocktail (Sigma). • Remove the supernatant carefully using a 1 ml pipette, leaving about 50 µl. Note: Always use a pipette rather than decanting the supernatant and leave behind some liquid above the pellet to avoid contamination. Subsequently, equation 2, 3, or 4 was chosen to calculate the pellet volume depending on the geometry of the container. During phase separation, chemicals are converted from a matrix or an aqueous medium to a solvent (for additional chemical or molecular biological analysis). Pellet volumes were measured after the different centrifugations in JA14 and JA18 rotor containers. Do not aliquot the antibody.For western blots, incubate membrane with diluted primary antibody in 5% w/v BSA, 1X TBS, 0.1% TweenFrom sample preparation to detection, the reagents you need for your Western Blot are now in one convenient kit: This protocol is intended for immunoprecipitation of native proteins for analysis by western immunoblot or kinase activity utilizing Protein A magnetic separation.A cell lysate pre-clearing step is highly recommended to reduce non-specific protein binding to the Protein A Magnetic beads. Suspend cell pellet in 5 ml of complete spinner medium-5 by pipetting up and down to disrupt clumps. By continuing you agree to the Copyright © 2020 Elsevier B.V. or its licensors or contributors. Supernatant in the largest biology dictionary online. Fix for 15 min at room temperature (20-25°C). Initial studies indicated that RCAS1 was secreted from cancer cells and functioned as a ligand for a putative receptor expressed on NK cells, as well as T and B lymphocytes, inducing their apoptosis, which enabled cancer cells to evade immune surveillance (5,6). Centrifuge for 10 minutes at 2,000 x g at 4°C to separate debris and dead cells. Centrifuge at 13,000 rpm for 10 min at 4°C to pellet insoluble contents.