IKBKB KO cell lysate available now. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 14 bp deletion in exon 6.
I-kappa-B kinase 2, I-kappa-B-kinase beta, IKK-beta, IKK2, IKKB_HUMAN, IMD15, IkBKB, Inhibitor of kappa light polypeptide gene enhancer in B cells, kinase beta, Inhibitor of nuclear factor kappa-B kinase subunit beta, NFKBIKB, Nuclear factor NF-kappa-B inhibitor kinase beta
IKBKB KO cell lysate available now. Free of charge wild type control included. Knockout achieved by using CRISPR/Cas9, Homozygous: 14 bp deletion in exon 6.
Western blot data indicates that the CRISPR gene edit may have resulted in a truncation of the protein of interest. Please see data images.
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Lysate preparation: Our lysates are made using RIPA buffer to which we add a protease inhibitor cocktail and phosphatase inhibitor cocktail (ratio: 300:100:10). This means that the protein of interest is denatured. If you require a native form of the protein please use the live cell version. Please refer to our lysis protocol for further details on how our lysates are prepared.
User storage instructions: Lyophilizate may be stored at 4°C. After reconstitution, store at -20°C for short-term storage or -80°C for long-term storage.
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Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: IKBKB knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - Anti-IKK beta antibody [EPR6043] ab124957 observed at 90 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-IKK beta antibody [EPR6043] ab124957 Anti-IKK beta antibody [EPR6043] was shown to specifically react with IKK beta in wild-type HeLa cells in western blot. The band observed in the knockout cell line Human IKBKB (IKK beta) knockout HeLa cell line ab264847 (knockout cell lysate ab257228) lane below 90kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and IKK beta knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-IKK beta antibody [EPR6043] ab124957 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 500 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-IKK beta antibody [EPR6043] (Anti-IKK beta antibody [EPR6043] ab124957) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: IKBKB knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 87 kDa
Observed band size: 90 kDa
Lane 1: Wild-type HeLa cell lysate (20µg)
Lane 2: IKBKB knockout HeLa cell lysate (20µg)
Lanes 1- 2: Merged signal (red and green). Green - Anti-IKK beta antibody [Y466] ab32135 observed at 87 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245 observed at 37 kDa.
Anti-IKK beta antibody [Y466] ab32135 Anti-IKK beta antibody [Y466] was shown to specifically react with IKK beta in wild-type HeLa cells in western blot. The band observed in the knockout cell line Human IKBKB (IKK beta) knockout HeLa cell line ab264847 (knockout cell lysate ab257228) lane below 87kDa may represent truncated forms and cleaved fragments. This has not been investigated further. Wild-type and IKK beta knockout samples were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. Anti-IKK beta antibody [Y466] ab32135 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) were incubated overnight at 4°C at 1 in 1000 and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-IKK beta antibody [Y466] (Anti-IKK beta antibody [Y466] ab32135) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: IKBKB knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 87 kDa
Observed band size: 87 kDa
Homozygous: 14 bp deletion in exon 6
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