TGFBR2 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift = 97%.
AAT3, FAA3, LDS1B, LDS2, LDS2B, MFS2, RIIC, TAAD2, TGF beta receptor type IIB, TGF-beta receptor type II, TGF-beta receptor type-2, TGF-beta type II receptor, TGF-beta-R2, TGFB R2, TGFR2_HUMAN, TGFbeta - RII, TbetaR-II, Transforming growth factor beta receptor II, Transforming growth factor beta receptor type IIC, Transforming growth factor, beta receptor II (70/80kDa), Transforming growth factor-beta receptor type II, transforming growth factor, beta receptor II alpha, transforming growth factor, beta receptor II beta, transforming growth factor, beta receptor II delta, transforming growth factor, beta receptor II epsilon, transforming growth factor, beta receptor II gamma
TGFBR2 KO cell line available to order. KO validated by Next Generation Sequencing, Western blot. Free of charge wild type control provided. Knockout achieved by CRISPR/Cas9 X = 1 bp insertion Frameshift = 97%.
Upon arrival, the vial should be stored in liquid nitrogen vapor phase and not at -80°C. Storage at -80°C may result in loss of viability.
1. Thaw the vial in 37°C water bath for approximately 1-2 minutes.
2. Transfer the cell suspension (0.8 mL) to a 15 mL/50 mL conical sterile polypropylene centrifuge tube containing 8.4 mL pre-warmed culture medium, wash vial with an additional 0.8 mL culture medium (total volume 10 mL) to collect remaining cells, and centrifuge at 201 x g (rcf) for 5 minutes at room temperature. 10 mL represents minimum recommended dilution. 20 mL represents maximum recommended dilution.
3. Resuspend the cell pellet in 5 mL pre-warmed culture medium and count using a haemocytometer or alternative cell counting method seed all remaining cells into a T25.
4. Incubate the culture at 37°C incubator with 5% CO2. Check the culture one day after revival and continue to check until 80% confluent. Media change can be given if needed.
5. Once confluent passage into an appropriate flask at a density of 2x104 cells/cm2. Seeding density is given as a guide only and should be scaled to align with individual lab schedules. Cultures should be monitored daily.
We will provide viable cells that proliferate on revival.
This product is subject to limited use licenses from The Broad Institute, ERS Genomics Limited and Sigma-Aldrich Co. LLC, and is developed with patented technology. For full details of the licenses and patents please refer to our limited use license and patent pages.
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Full details and terms and conditions can be found here:
Terms & Conditions.
Anti-TGF beta Receptor II antibody [EPR14673] ab184948 was shown to specifically react with in wild-type A549 cells as signal was lost in TGFBR2 knockout cell line ab261863 (knockout cell lysate Human TGFBR2 (TGF beta Receptor II) knockout A549 cell lysate ab261672). Wild-type and TGFBR2 knockout samples were subjected to SDS-PAGE. Anti-TGF beta Receptor II antibody [EPR14673] ab184948 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4° at 1/2000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-TGF beta Receptor II antibody [EPR14673] (Anti-TGF beta Receptor II antibody [EPR14673] ab184948) at 1/2000 dilution
Lane 1: Wild-type A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 2: TGFBR2 knockout A549 (Human lung carcinoma cell line) whole cell lysate at 20 µg
Lane 2: Western blot - Human TGFBR2 (TGF beta Receptor II) knockout A549 cell line (ab261863)
Lane 3: HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate at 20 µg
Lane 4: HT-1080 whole cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 65 kDa
Observed band size: 80 kDa
X = 1 bp insertion
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