Primary antibodies against the following were used: HIF-1α (mouse monoclonal, 1:1 500, MAB5382; Chemicon Inc, Billerica, MD); BNIP3 (mouse monoclonal, 1:3000, B7931; Sigma, St. Louis, MI); BCL-xL (rabbit polyclonal, SAB3500349; 1:1000, Sigma, St. Louis, MI); VEGF (rabbit polyclonal, 1:500, PB0084; Boster, Wuhan, China); GAPDH (mouse monoclonal, 1:10000, KC-5G4; KangCheng, Shanghai, China); and β-tubulin (mouse monoclonal, 1:1000, A06868; Boster, Wuhan, China). Horseradish peroxidase-labeled secondary antibodies were obtained from Zymed Laboratories Inc (San Francisco, CA). Western blot assays were performed as previously described [16 (link)].
β tubulin
Manufactured by Boster Bio
Sourced in China, United States
β-Tubulin is a key structural protein found in the cytoskeleton of eukaryotic cells. It is an essential component of microtubules, which play crucial roles in various cellular processes such as cell division, intracellular transport, and cell motility.
Automatically generated - may contain errors
Lab products found in correlation
P ampk, by Cell Signaling Technology (2 mentions)
Bcl xl, by Merck Group (1 mentions)
Bnip3, by Merck Group (1 mentions)
Horseradish peroxidase labeled secondary antibodies, by Zymo Research (1 mentions)
Mab5382, by Merck Group (1 mentions)
B7931, by Merck Group (1 mentions)
Pb0084, by Boster Bio (1 mentions)
Vascular endothelial growth factor (vegf), by Boster Bio (1 mentions)
Hrp conjugated anti rabbit and anti mouse secondary antibodies, by Boster Bio (1 mentions)
Lipopolysaccharides (lps), by Merck Group (1 mentions)
Trizol, by Merck Group (1 mentions)
Compound c, by Selleck Chemicals (1 mentions)
P ulk, by Cell Signaling Technology (1 mentions)
3 methyladenine 3 ma, by Selleck Chemicals (1 mentions)
P erk, by Cell Signaling Technology (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
Leptin, by Santa Cruz Biotechnology (1 mentions)
Pgc 1α, by Santa Cruz Biotechnology (1 mentions)
Pparγ, by Abcam (1 mentions)
Cyclin b, by Santa Cruz Biotechnology (1 mentions)
6 protocols using β tubulin
1
Evaluating Hypoxia Signaling Pathways
2
Autophagy Regulation in Cell Signaling
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TRIzol, LPS and phenylmethanesulfonyl fluoride (PMSF) were purchased from Sigma (Saint Louis, MO, USA). Deh was purchased from Shanghai Yuanye Bio-Technology Co. CCK8 was purchased from Saint-Bio Co. (Shanghai, China). Compound C (CC) and 3-methyladenine (3-MA) were purchased from Selleckchem (Shanghai, China). p-Beclin, Beclin, p-ULK, ULK, AMPK, p-AMPK and LC3B were purchased from Cell Signaling Technology (Boston, MA, USA). P62 was purchased from Proteintech (Rosemont, IL, USA). β-Tubulin was purchased from Bosterbio in Dallas (Pleasanton, USA). IgG was purchased from Beyotime (Shanghai, China). HRP-conjugated anti-mouse and anti-rabbit secondary antibodies were purchased from Bosterbio.
3
Western Blot Analysis of Cellular Signaling
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Protein extraction and Western blot analysis of tissues and cells were performed according to our previous methods [14 (link)]. Antibodies against PCNA, cyclin B, cyclin D1, cyclin E, p27, leptin, and PGC-1α were from Santa Cruz Biotechnology. Antibodies against p-PI3K, PI3K, p-AKT, AKT, p-mTOR, mTOR, p-ERK, ERK, p-p38, and p38 were from Cell Signaling Technology. Antibodies against FASN (BOSTER, wuhan, China), FABP4 (BOSTER, China), PPARγ (abcam, Cambridge, MA, USA), and β-tubulin (Boster, China) were used. BODIPY staining was performed according to our published method; the intensity of fluorescence was analyzed by ImageJ [14 (link)].
4
Western Blot Analysis of Protein Levels
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Liver cancer cell lines and gastric cancer cell lines with a culture density of approximately 80% were washed twice with phosphate-buffered saline (PBS) buffer. Cells were completely lysed with phenylmethylsulfonyl fluoride-spiked high-performance lysis solution (RIPA Lysis Buffer) (Boster, Wuhan), centrifuged (12000 × rpm, 4 ℃, 10 min) and the supernatant collected. Protein concentration was measured using a BCA protein assay kit (Solarbio, Beijing). Each protein sample of 20 μg was separated by SDS-PAGE and transferred to polyvinylidene difluoride membranes (Millipore, United States). The membranes were subsequently closed with protein-free fast closure solution (Boster, Wuhan) and rinsed three times for 5 min each time with TBST rinse solution. The membranes were incubated with primary antibodies of PDHB (Proteintech Group, Wuhan) and β-Tubulin (Boster, Wuhan) overnight on a 4 ℃ shaker, followed by incubation with secondary antibodies for 1 h, three rinses with TBST rinse solution for 10 min each time and then with an enhanced chemiluminescence kit (Boster, Wuhan) to visualize the blots. The total grey values of the protein bands were analyzed using ImageJ software to quantify the protein expression levels of the genes.
5
Immunoblotting Signaling Pathway Proteins
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AMPK (1:1000, CAT: 5832S), p-AMPK (1:1000, CAT: 2535S), ERK1/2 (1:1000, CAT: 4695S), p-ERK1/2 (1:1000, CAT: 9101L), JNK1/2 (1:1000, CAT: 9252S), p-JNK1/2 (1:1000, CAT: 9251L), P38 (1:1000, CAT: 9212S), p-P38 (1:1000, CAT: 4631S) and NF-κB-p65 (1:1000, CAT: 8242S) were purchased from Cell Signaling Technology (Boston, MA, USA), P62 (WB: 1:1000, Co-IP: 1:50, CAT: 66184-2-2g), LC3B (1:1000, CAT: 18725-1-AP) and Keap1 (1:1000, CAT: 10503-2-AP) were purchased from Proteintech (Rosemont, IL, USA), HO-1(1:1000, CAT: GK284419-11), NRF-2 (WB: 1:1000, IF: 1:150, CAT: Gk298097-1) were purchased from Abcam (Cambridge, UK), GPR109A (WB: 1:200, IHC: 1:50, CAT: NBP1-92180) were purchased from Novus Biologicals (Shanghai, China), β-Tubulin (1:6000, CAT: M05613-4) were purchased from Bosterbio (Pleasanton, CA, USA), IgG (Co-IP: 1 μg, CAT:A7028) were purchased from Beyotime (Shanghai, China), (HRP)-conjugated anti-mouse (1:10,000, CAT: BA1051) or anti-rabbit secondary antibodies (1:10,000, CAT: BA1055) were purchased from Bosterbio (CA, USA).
6
Western Blot Analysis of NLRP3 Inflammasome
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Protein samples were collected and prepared. After electrophoresis and membrane transfer, the sample was incubated with primary antibodies. The primary antibodies were NLRP3 (1:1000; #ab91413; Abcam), caspase‐1 (1:500; #sc‐56036; Santa), ASC (1:1000; #sc‐514414; Santa), cathepsin B (1:1000; CST, #31718), β‐actin(1:1000; #BM0627; BOSTER, China), and β‐tubulin (1:1000; #A05397‐1; BOSTER). And then, membranes were treated with anti‐rabbit IgG (1:1500; #5127; CST) or anti‐mouse IgG (1:1500; #93702; CST). All protein bands were quantitated by Image J software (NIH, Bethesda, USA).
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