Western blotting was performed as described previously (14 (link)). The following primary antibodies were used: MRPL9 (15342-1-AP, 1:1000, Proteintech, China), β-Tubulin (10068-1-AP, 1 : 1,000, Proteintech, China), E-cadherin (24E10, 1:800, CST, US), β-catenin (sc-7199, 1:800, Sants Cruz Biotechnology, US), N-cadherin (22018-1-AP, 1:1000, Proteintech, China). The protein signals were visualized using ECL ((Millipore, Massachusetts, Uinted States) and the signal strength was analyzed using Image Lab V6.0 software.
β tubulin 10068 1 ap
Manufactured by Proteintech
Sourced in China
β-Tubulin (10068-1-AP) is a primary antibody that specifically recognizes the β-tubulin protein. β-Tubulin is a component of microtubules, which are cytoskeletal structures involved in various cellular processes such as cell division, intracellular transport, and the maintenance of cell shape.
Automatically generated - may contain errors
Lab products found in correlation
Gapdh 60004 1 ig, by Proteintech (2 mentions)
Enhanced chemi luminescence (ecl), by Merck Group (1 mentions)
N cadherin, by Proteintech (1 mentions)
Protease inhibitor cocktail, by Apexbio (1 mentions)
Ipvh00010, by Merck Group (1 mentions)
Xirp1 sc 166658, by Santa Cruz Biotechnology (1 mentions)
Bca protein assay kit, by Tiangen Biotech (1 mentions)
Alexa fluor 647 goat anti mouse a0473, by Beyotime (1 mentions)
Tlr4 66350 1 ig, by Proteintech (1 mentions)
Lipopolysaccharides l4391, by Merck Group (1 mentions)
Goat anti rabbit igg h l, by Cell Signaling Technology (1 mentions)
Alexa fluor 488 goat anti rabbit a0423, by Beyotime (1 mentions)
M mlv reverse transcriptase, by Promega (1 mentions)
Qiaamp rna blood mini kit, by Qiagen (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Goat anti mouse igg as003, by ABclonal (1 mentions)
4 protocols using β tubulin 10068 1 ap
1
Protein Expression Analysis by Western Blot
2
Protein Extraction and Western Blot Analysis
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Cells or heart tissue were lysed with RIPA buffer (50 mM Tris pH 7.4, 150 mM NaCl, 1% Triton X-100, 1% sodium deoxycholate, 0.1% SDS) (Beyotime Biotechnology, P0013B) with protease inhibitor cocktail (APExBIO, K1012). Protein concentrations were determined by BCA Protein Assay Kit (TIANGEN, PA115). Equal amounts of protein were loaded on 10% SDS-PAGE. After electrophoresis, proteins were transferred to PVDF membrane (Millipore, IPVH00010). After blocked with 5% skimmed milk at room temperature, membranes were incubated with primary antibodies (Xirp1: sc-166658, Santa Cruz Biotechnology, diluted at 1:1,000; GAPDH: 60,004-1-Ig, Proteintech, diluted at 1:1,000; β-tubulin: 10068-1-AP, Proteintech, diluted at 1:2,000) at 4°C overnight and then incubated with corresponding secondary antibody. Western blots results were detected with Tanon 5200 Multi.
3
Antibody-Based Signaling Pathway Analysis
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Antibodies: AKT (4691), P38 (8690), ERK1/2 (4695), p-AKT (4060), p-P38 (4511), p-ERK1/2 (4370), IκBα (4814), p-IκBα (2859), p-IKKα/β (2697), NF-κB (8242), Rab5a (46449), horse anti-mouse IgG (H&L) (7076), and goat anti-rabbit IgG (H&L) (7074) were purchased from Cell Signaling Technology (Danvers, MA, United States). IKKβ (ab124957), PBR (ab109497) was from Abcam (CA, United States). Alexa Fluor 488 goat anti-rabbit (A0423) and Alexa Fluor 647 goat anti-mouse (A0473) secondary antibodies were obtained from Beyotime Biotechnology (Shanghai, China). TLR4 (66350-1-Ig), GAPDH (60004-1-Ig), and β-Tubulin (10068-1-AP) were purchased from Protein-Tech (Wuhan, Hubei, China). Lipopolysaccharides(L4391) was obtained from Sigma (Louis, MO, United States), Remimazolam Tosilate for Injection (Remimazolam) were obtained from Hengrui (Jiangsu, China).
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4
Isolation and Characterization of Human Norovirus GII.4
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The sequence of HuNoV GII.4 used here (Gene bank: OL721917) was isolated from positive stool samples of diarrhea patients [8 (link)]. Briefly, QIAamp RNA Blood Mini Kit (Qiagen, 52304, Hilden, Germany) was used to isolate the HuNoV genome from the positive stool sample, and then moloney murine leukemia virus (M-MLV) reverse transcriptase (Promega, M170B, Dane County, WI, USA) was used to synthesize the HuNoV cDNA. The progeny viruses were obtained by transfecting HEK293T cells with the plasmid, which encoded the full-length cDNA of HuNoV. Virus stocks were aliquoted and stored at −80 °C until use, and the genome copy was determined. Human colon epithelial cell lines Caco2 cells and HEK293T cells were purchased from ATCC and cultured in Dulbecco’s modified eagle’s medium (DMEM) containing 10% fetal bovine serum (FBS), purchased from Thermo Scientific, Sydney, Australia, and 100 U/mL penicillin/streptomycin (Genom, Hangzhou, China) at 37 °C with 5% CO2 in an incubator. Antibodies against NLRP3 (19771-1-AP), caspase1 (22915-1-AP), N-GSDMD (20770-1-AP), and β-tubulin (10068-1-AP) were purchased from Proteintech, Wuhan, China. Besides, HRP-conjugated goat anti-rabbit IgG (AS063) or goat anti-mouse IgG (AS003) were also purchased from Abclonal, Wuhan, China.
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