Proteins were isolated from ovine SVFs using a lysis buffer supplemented with protease inhibitors (Solarbio, Beijing, China), phosphatase inhibitors (Solarbio, Beijing, China), and phenylmethylsulfonyl fluoride (PMSF, Solarbio, Beijing, China). Equal amounts of protein were separated by SDS-PAGE, transferred to nitrocellulose filter membranes (Solarbio, Beijing, China), and sealed in 5% skim milk for 1 h. Membranes were then incubated with primary antibodies (anti-β-actin: Immunoway, Beijing, China; anti-MEOX2: Proteintech, Wuhan, China; anti-adiponectin: Proteintech, Wuhan, China; anti-C/EBPα: Proteintech, Wuhan, China; anti-FABP4: Proteintech, Wuhan, China; anti-PPARγ: Proteintech, Wuhan, China) and secondary antibodies (LI-COR, Lincoln, NE, USA). After washing three times, the membranes were imaged using an Odyssey Clx imaging system (LI-COR, Lincoln, NE, USA).
Anti β actin
Manufactured by Immunoway
Sourced in China, United States
Anti-β-actin is a laboratory reagent used to detect and quantify the expression of the β-actin protein, which is a widely expressed and abundant cytoskeletal protein. It is commonly used as a reference or control in various analytical techniques, such as Western blotting, immunohistochemistry, and flow cytometry, to normalize and compare protein expression levels across samples.
Automatically generated - may contain errors
Lab products found in correlation
Protease inhibitor, by Solarbio (3 mentions)
Phosphatase inhibitor, by Solarbio (3 mentions)
Odyssey clx imaging system, by LI COR (3 mentions)
Anti bcl 2, by Immunoway (2 mentions)
Nitrocellulose membrane, by Solarbio (2 mentions)
Anti hsd17b12, by Thermo Fisher Scientific (2 mentions)
Image pro plus software, by Media Cybernetics (2 mentions)
Ripa buffer, by Solarbio (2 mentions)
Phenylmethylsulfonyl fluoride, by Solarbio (2 mentions)
Bicinchoninic acid protein assay kit, by Thermo Fisher Scientific (2 mentions)
Ab7090, by Abcam (2 mentions)
Nitrocellulose membrane, by Merck Group (2 mentions)
Ripa lysis buffer, by Meilun (2 mentions)
Anti collagen 1, by Abcam (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Nitrocellulose filter membranes, by Solarbio (1 mentions)
Anti adiponectin, by Proteintech (1 mentions)
Anti c ebpα, by Proteintech (1 mentions)
Anti fabp4, by Proteintech (1 mentions)
Anti pparγ, by Proteintech (1 mentions)
15 protocols using anti β actin
1
Western Blot Analysis of Ovine Stromal Vascular Fraction Proteins
2
Stattic Modulates Apoptosis Markers in CD133+ Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Sorted CD133+ cells were treated with McCoy's 5A containing 0 μM, 0.625 μM, or 1.25 μM stattic for 24 h. Thereafter, protein lysates were collected and a protein assay (BioRad) was performed. The concentration of total protein in cells was quantified using the bicinchoninic acid (BCA) protein assay reagent following the manufacturer's instructions. Cells were lysed with MPER (Pierce) supplemented with protease and phosphatase inhibitors for 30 min on ice, and lysates were clarified by centrifugation for 5 min at 12,000 /g. Twenty-five to fifty milligrams of lysates were loaded onto 5% SDS-PAGE gels and were transferred onto PVDF membranes. The membranes were blocked with 5% skim milk for 1 h and were incubated overnight at 4°C with anti-Bcl-2 (ImmunoWay, USA), anti-Bcl-xl (ImmunoWay, USA), anti-P-STAT3 (ImmunoWay, USA), anti- STAT3 (ImmunoWay, USA) antibody, anti-β-actin (ImmunoWay, USA) antibody (1/1000 dilution) was used as the internal loading control. The membranes were washed three times and then were incubated with a secondary antibody (1/1000 dilution) at room temperature for 1 h. Finally, the membranes were washed three times and were exposed in the dark room; ImageJ software was used for image analysis.
3
Western Blot Analysis of BMPER
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
WB was carried out as described previously [4 (link)–6 (link)]. Anti-BMPER and anti-β-actin antibodies were purchased from ImmunoWay (USA) and Wanleibio (Shenyang, China).
4
Western Blot Analysis of TRPV1, NF-κB p65
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein concentration was quantified using a BCA kit (Beyotime, China). Equivalent amounts of protein were separated by 12% SDS-PAGE and then transferred onto PVDF membranes. The membrane was blocked by 5% BSA in TBST for 1 h. The membrane was incubated with primary antibodies against TRPV1 (1 : 1,000, Abcam, UK), NF-κB p65 (1 : 1,000, Proteintech, China), and anti-β-actin (1 : 2,000, ImmunoWay, USA) at 4°C overnight. After washing, membranes were incubated with HRP-coupled secondary antibodies (1 : 2,000, Zhongshan Golden Bridge, China). After washing with TBST, immunoblots were visualized with ECL-Plus kit (Beyotime, China). Grey values of immunoreactive bands were normalized to β-actin in each sample.
5
Western Blot Analysis of Cellular Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blotting was carried out as described before.[35 ] Image Lab 4.0 software (Bio‐Rad) was used for the analysis of blots. Primary antibodies used for western blotting were listed as follows: anti‐BCAT1 (1:1000, Proteintech, 13640‐1‐AP), anti‐β‐Tubulin (1:2000, Immunoway, YT4780), anti‐GPX4 (1:1000, ABclonal, A11243), anti‐β‐actin (1:2000, Immunoway, YM3028), anti‐H3K9me3 (1:2000, ABclonal, A2360), anti‐Histone H3 (1: 2000, ABclonal, A2348), anti‐EGR1 (1:2000, ABclonal, A2722), anti‐HSPB1 (1:1000, ABclonal, A16332), anti‐HSF1 (1:1000, ABclonal, A13765), anti‐FLAG (1:2000, ABclonal, AE005), anti‐PTGS2 (1:1000, ABclonal, A1253), anti‐SLC7A11 (1:1000, ABclonal, A2413), anti‐ATG5 (1:1000, ABclonal, A19677), and anti‐ATG7 (1:1000, ABclonal, A19604).
6
Comprehensive Western Blot Analysis of Key Cellular Regulators
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analysis was performed as previously described [63 (link)]. The primary antibodies used in this study were anti-β-actin (immunoway), anti-NR4A1 (immunoway), anti-METTL3 (abcam), anti-YTHDF1(Saier Biotechnology), anti-YTHDF2 (abcam), anti-YTHDF3(Saier Biotechnology), anti-AKT1 (Saier Biotechnology), anti-p-AKT1(S473) (Wanlei Biotechnology), anti-DDX6 (Saier Biotechnology), anti-LSD1 (Saier Biotechnology), anti-CoREST (Saier Biotechnology), anti-HDAC1 (Saier Biotechnology), anti-EGFP (Saier Biotechnology), anti-Vimentin (abcam), anti-N-Cadherin (abcam), anti-SP1 (abcam), anti-E- Cadherin (abcam) and anti-Flag (MBL). β-actin was the internal reference.
7
Huaier Modulates Cellular Signaling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were treated with a series concentrations of Huaier (0, 5, 10 and 15mg/ml) for 24h, and then lysed by cell lysis buffer for Western and IP (Beyotime, P0013). Protein concentration was detected with BCA Protein Assay Kit (KeyGen BioTECH, KGP902). After that, 20µg proteins were separated by 10% or 12% SDS-PAGE and transferred onto Pure Nitrocellulose Blotting Membrane (Pall Corporation, P/N 66485). Then, the blots were blocked for nonspecific binding with 5% non-fat milk in PBST (PBS, Tween-20, pH7.4) at room temperature for 1h. Next, the blots were incubated overnight at 4℃ with 5% non-fat milk containing primary antibodies which are listed as follows: anti-PCNA (ImmunoWay, YM3031), anti-Ki-67 (ImmunoWay, YT2467), anti-β-actin (ImmunoWay, YM3028), anti-Bcl-2 (ImmunoWay, YM3041), anti-Bax (ImmunoWay, YT0455), anti-E-cadherin (ImmunoWay, YT1454), anti-N-cadherin (ImmunoWay, YT2988), anti-YAP1 (abcam, ab52771), anti-p-YAP1 (ab76252), anti-CyclinD1 (abcam, ab134175), anti-Cleaved Caspase Substrate Motif (Cell Signaling, #8698) and anti-YAP/TAZ (Cell Signaling, #8418). After that, incubating blots with secondary antibodies conjugated with HRP (Cell Signaling, #7074 or #7076) for 1h at room temperature. Finally, after washing three times with PBST, the blots were visualized by New Super ECL Assay (KeyGen BioTECH, KGP1128).
8
Western Blot Analysis of HSD17B12
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total proteins were collected from the cell samples after lysis with a lysis buffer (RIPA buffer, Solarbio, Beijing, China) supplemented with protease inhibitors (Solarbio, Beijing, China), phosphatase inhibitors (Solarbio, Beijing, China), and phenylmethylsulfonyl fluoride (Solarbio, Beijing, China). Sodium dodecyl sulfate–polyacrylamide gel electrophoresis was performed on a 10% gel. After filtration through a nitrocellulose membrane (Solarbio), the membrane was sealed with 5% skim milk and treated with primary antibodies (anti-HSD17B12, 1:500, Thermo Fisher Scientific, Shanghai, China; anti-β-actin, 1:3000, Immunoway, Beijing, China) and secondary antibodies. Finally, the membranes were imaged using an Odyssey Clx Imaging System (LICOR, Lincoln, NE, USA) and quantified using Image-Pro Plus software (Media Cybernetics, Rockville, MD, USA).
9
Western Blot Analysis of CTSH Protein
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Using a RIPA lysis buffer (Meilun Biotechnology, China), total protein was extracted from cells. A bicinchoninic acid protein assay kit (#23227; Thermo Fisher Scientific, Waltham, USA) was used to measure protein concentration. Denatured proteins were subject to 10% SDS-PAGE and blotted onto nitrocellulose membranes (Millipore, Bedford, USA). After being blocked with 5% skimmed dry milk for 2 h, the membranes were subsequently incubated with primary antibodies, including anti-CTSH (1:1000, Immunoway), and anti-β-Actin (1:1000, Immunoway) overnight at 4 °C, then incubation with horseradish peroxidase labeled secondary antibodies (ab7090, 1:5000; Abcam) followed. β-Actin was used to normalize the expression of target proteins.
10
Western Blot Analysis of HSD17B12
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total proteins were collected from the cell samples after lysis with a lysis buffer (RIPA buffer, Solarbio, Beijing, China) supplemented with protease inhibitors (Solarbio, Beijing, China), phosphatase inhibitors (Solarbio, Beijing, China), and phenylmethylsulfonyl fluoride (Solarbio, Beijing, China). Sodium dodecyl sulfate–polyacrylamide gel electrophoresis was performed on a 10% gel. After filtration through a nitrocellulose membrane (Solarbio), the membrane was sealed with 5% skim milk and treated with primary antibodies (anti-HSD17B12, 1:500, Thermo Fisher Scientific, Shanghai, China; anti-β-actin, 1:3000, Immunoway, Beijing, China) and secondary antibodies. Finally, the membranes were imaged using an Odyssey Clx Imaging System (LICOR, Lincoln, NE, USA) and quantified using Image-Pro Plus software (Media Cybernetics, Rockville, MD, USA).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!