The CDK9 inhibitor, LDC000067, was purchased from Selleck (Houston, TX). LDC000067 was dissolved in dimethyl sulfoxide (DMSO) for in vitro experiments and in CMC-Na (1%) for in vivo study. Atorvastatin was purchased from Pfizer Ireland Pharmaceuticals (New York, USA). Antibodies against NF-κB p65 (sc-8008), Lamin B (sc-56144) and GAPDH (sc-32233) were from Santa Cruz Biotechnology (Santa Cruz, CA, USA); antibodies against CDK9 (2316S), p-CDK9 (2549S) and IκB (4814S) were from Cell Signaling Technology (Danvers, MA); antibodies against α-SMA (ab32575), Vimentin (ab8978), OPN (ab8448), PCNA (ab29), β-actin (ab8227) and CD68 (ab955) were from Abcam (Cambridge, MA, USA). Mouse CDK9 ELISA kit (E-EL-M0378c) was from Elabscience Biotechnology Co., Ltd (Wuhan, China).
Iκb 4814s
Manufactured by Cell Signaling Technology
Sourced in United States
IκB (4814S) is a product offered by Cell Signaling Technology. It is a laboratory equipment used in research applications. The core function of this product is to detect IκB, a regulatory protein that inhibits the activity of NF-κB transcription factors.
Automatically generated - may contain errors
Lab products found in correlation
Gapdh sc 32233, by Santa Cruz Biotechnology (1 mentions)
Ldc000067, by Selleck Chemicals (1 mentions)
Ab8227, by Abcam (1 mentions)
Ab955, by Abcam (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
P65 af5006, by Affinity Biosciences (1 mentions)
P p65 af2006, by Affinity Biosciences (1 mentions)
Imagej software, by Media Cybernetics (1 mentions)
Myd88, by ABclonal (1 mentions)
2 protocols using iκb 4814s
1
CDK9 Inhibitor LDC000067 Protocol
2
Ruminal Epithelial Protein Extraction and Western Blot Analysis
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Total protein from ruminal epithelial cells was extracted using RIPA lysis buffer and quantified using a BCA Protein Assay kit (P0012, Beyotime Biotechnology, China). Protein was separated by SDS-PAGE on 10% gels and transferred to PVDF membranes. Then the membrane was Blocking with 5% skimmed milk or BSA in TBST buffer for 2 h, and followed incubated with TLR4 (ab217274, abcom, Cambridge, the United Kingdom), MyD88 (A0980, Abclonal, Wuhan, Chian), p-p65 (AF2006, Affinity, Jiangsu, China), p65(AF5006, Affinity, Jiangsu, China), p-IκB (9246S, Cell Signaling Technology, Danvers, MA, USA), IκB(4814S, Cell Signaling Technology, Danvers, MA, USA ). antibodies overnight at 4°C, subsequently incubated for 2 h at room temperature with secondary antibodies. The target bands were detected using the chemiluminescence (ECL) system and analyzed using the ImageJ software (Media Cybernetics, Bethesda, MD, United States).
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