The selective IKKβ inhibitors IKKβIII (401480) and IKKβV (401482) were purchased from Calbiochem (Dorset, UK) and dissolved in dimethyl sulfoxide (DMSO) for in vitro and ex vivo studies. All solvents and reagents were purchased from Sigma-Aldrich (Dorset, UK) unless otherwise stated. Tissue culture medium was obtained from Invitrogen (Paisley, UK). Human macrophage colony-stimulating factor (M-CSF, 416-ML-050) was purchased from R&D Systems (Abingdon, UK). RANKL was a gift from Patrick Mollat (Galapagos SASU, France) [16 (link)]. Foetal bovine serum (FBS) was obtained from GE Healthcare. Vitamin C (ascorbic acid) was obtained from BDH Laboratory Supplies (Dorset, UK). The following antibodies were used for Western blotting and immunostaining: anti-pIκB (ser32, 2859), anti-IκB (4812) from Cell Signalling Technology (Boston, MA, USA) and anti-β-actin (A5060) from Sigma-Aldrich (Dorset, UK).
Anti β actin a5060
Manufactured by Merck Group
Sourced in United States, United Kingdom
Anti-β-actin (A5060) is a primary antibody produced in rabbit that specifically binds to the β-actin protein. β-actin is a widely expressed cytoskeletal protein that is commonly used as a reference or loading control in various biological assays.
Automatically generated - may contain errors
Lab products found in correlation
Foetal bovine serum (fbs), by GE Healthcare (1 mentions)
Nitrocellulose, by Cytiva (1 mentions)
Anti chk1, by Cell Signaling Technology (1 mentions)
Enhanced chemi luminescence (ecl), by Cell Signaling Technology (1 mentions)
Pser345 chk1, by Cell Signaling Technology (1 mentions)
Pthr68 chk2, by Cell Signaling Technology (1 mentions)
Okadaic acid, by Merck Group (1 mentions)
2 protocols using anti β actin a5060
1
Selective IKKβ Inhibition Protocol
2
Cell Lysis and Protein Analysis
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Whole-cell extracts were prepared as described previously.44 (link) The cell lysis buffer was supplemented with 500 nM okadaic acid (Merck Chemicals, VWR International, Beeston, UK) when samples were used to analyse Aurora kinase sites of phosphorylation. Proteins were separated by SDS-PAGE and blotted onto nitrocellulose (Schleicher & Schuell). Proteins were detected using the enhanced chemiluminiscence detection system (ECL, Amersham) according to the manufacturer's recommendations using anti-CHK1 (#2345), pAurora: Phospho-Aurora A (Thr288)/Aurora B (Thr232)/Aurora C (Thr198) (#2914), p21 (#2947), pSer345 CHK1 (#2348), CHK1 (#2360), pThr68 CHK2 (#2661), CHK2 (#2662), pTyr15 CDK1 (#9111) from Cell Signaling, Hitchin, UK; anti-CDC45 (sc-20685), CDK1 (sc-54), Cyclin B1 (sc-245), pSer10 H3 (sc-8656), p53 (sc-126) from Santa Cruz Biotechnology (Wembley, UK) and anti-RPA34 (NA19L) from Calbiochem (Feltham, UK); or anti-β-actin (A-5060) from Sigma-Aldrich (Poole, UK).
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