Rat heart tissues and cell lysates were prepared using RIPA lysis buffer containing protease inhibitor (c1053-100, Applygen, China), and phosphatase inhibitors (p1260-1, Applygen, China). An equal amount of protein was separated on 10%–15% SDS-polyacrylamide gels and transferred onto PVDF membranes. After blocking with 5% nonfat milk at room temperature for 2 h, the PVDF membranes were incubated with the appropriate primary antibody, followed by a secondary antibody. The PVDF membrane was exposed to ECL reagents (p1020-25, Applygen, China) to visualize the protein signals. The antibodies used for Western blot were as follows: anti-PI3K (1F6A7, Proteintech, USA), anti-Akt (2C5D1, Proteintech, USA), anti-Phospho–PI3K (CY6427, Abways, China), anti-Phospho–Akt (P31749, Abways, China), anti-VEGFA (ab 214424, Abcam, USA), anti-ARG-1 (P05089, Abways, China), anti-mTORC1 (Q6UUV9, Abways, China), anti-CD206 (JF0953, HuaAn, China), anti-rabbit IgG H and L (ab0101, Abways, China), and anti-mouse IgG H and L (ab0102, Abways, China).
Ab0101
Manufactured by Abways
Sourced in China
The AB0101 is a laboratory centrifuge designed for general-purpose applications. It features a compact, durable construction and can accommodate a range of sample volumes and tube sizes. The centrifuge operates at adjustable speeds to meet various separation needs. Further details on the intended use or performance specifications are not available.
Automatically generated - may contain errors
Lab products found in correlation
Ab0102, by Abways (4 mentions)
Pvdf membrane, by Merck Group (3 mentions)
Ab0035, by Abways (2 mentions)
Ab214424, by Abcam (1 mentions)
C1053 100, by Applygen (1 mentions)
P1260 1, by Applygen (1 mentions)
Ab181602, by Abcam (1 mentions)
P0013, by Beyotime (1 mentions)
A12694, by ABclonal (1 mentions)
A16672, by ABclonal (1 mentions)
Wl03325, by Wanlei (1 mentions)
Wl01980, by Wanlei (1 mentions)
Ay0573, by Abways (1 mentions)
Chemidoc xr, by Bio-Rad (1 mentions)
A21010, by Abbkine (1 mentions)
Sc 271428, by Santa Cruz Biotechnology (1 mentions)
Sc 515931, by Santa Cruz Biotechnology (1 mentions)
Ab40772, by Abcam (1 mentions)
Ab76011, by Abcam (1 mentions)
Ab85936, by Abcam (1 mentions)
13 protocols using ab0101
1
Western Blot Analysis of Rat Heart
2
Protein Expression Analysis of Lichen Planus
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Four cases of fresh lichen planus lesion and normal skin tissues were collected and the total protein of tissue samples was lysed in cell lysis buffer (P0013; Beyotime) for western blot. Quantification of protein concentration was performed using BCA Protein Assay Kit (ZJ102, Shanghai Epizyme Biotechnology Co., Ltd.). The major antibodies used in western blot analysis are listed as follows: anti‐TAZ antibody TAZ (V386, Cell Signaling Technology, USA) (dilution 1:1000), anti‐GAPDH antibody (ab181602; Abcam, Cambridge, United Kingdom) (dilution 1:3000) and biotin‐labeled goat anti rabbit body IgG as secondary antibody (AB0101, Abways Technology, China). Western blot procedure was performed as previously described.
10 (link)
10 (link)
3
Protein Expression Analysis in Ischemic Brain
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The protein extracts of brain tissue were prepared as described previously. To determine NLRP3, ASC, and cleaved caspase-1 protein expressions in the ischemic core of the brain (Fig. 2 A), the protein concentrations were measured using BCA assay. Protein samples were separated by SDS-PAGE and were transferred onto polyvinylidene fluoride membranes. The membrane was blocked with 5% nonfat milk in TBST (0.1% Tween 20 in TBS) for 2 hours at room temperature and incubated with primary antibodies of anti-NLRP3 (1:1000, A12694; ABclonal Technology, Wuhan, China), anti-ASC (1:1000, A16672; ABclonal Technology, Wuhan, China), anti-cleaved caspase-1 (1:1000, WL03325; Wanleibio, Shenyang, China), and anti-NF-κB (1:500, WL01980; Wanleibio, Shenyang, China) at 4°C overnight. After being washed 3 times, the membranes were incubated with horseradish peroxidase-labeled goat anti-rabbit IgG (H + L) secondary antibodies (1:5000, AB0101; Abways Technology, Shanghai, China) for 2 hours. Then, the blots were visualized by enhanced chemiluminescence detection system (Chemidoc XRS+, Bio-Rad; Berkeley, CA). The results were normalized to anti-β-actin (1:1000, AY0573; Abways Technology, Shanghai, China) or anti-H3 (1:500, WL09804a; Wanleibio, Shenyang, China) and the intensity of bands was quantified by ImageJ.
4
Endometrial Protein Expression Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Endometrial tissues frozen at −80°C were homogenized in lysis buffer on ice, and the supernatant was quantified with bicinchoninic acid reagents. A total of 20 µg collected
protein was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto polyvinylidene difluoride (PVDF) membranes. After blocking with 5% BSA for
90 min at room temperature, the membranes were incubated with the primary antibodies integrin alpha V beta 3 (1:1,000; sc56-07, Novus Biologicals, Centennial, CO, USA), leukemia inhibitory
factor (LIF,1:1,000; sc-515931, Santa Cruz Biotechnology, Dallas, TX, USA), homeobox gene A10(HOXA10, 1:1,000; sc-271428, Santa Cruz
Biotechnology), and β-actin (1:10,000; 66009–1-Ig, Proteintech) overnight at 4°C. The membranes were then incubated with a horseradish peroxidase-conjugated goat anti-rabbit antibody
(1:5,000; AB0101, Abways, Shanghai, China) or goat anti-mouse antibody (1:10,000; A21010, Abbkine, Wuhan, China) at room temperature for 1 h. Protein bands were visualized using enhanced
chemiluminescence (ECL) and quantified with the ImageJ software (version 2.1; National Institutes of Health, Bethesda, MD, USA).
protein was separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto polyvinylidene difluoride (PVDF) membranes. After blocking with 5% BSA for
90 min at room temperature, the membranes were incubated with the primary antibodies integrin alpha V beta 3 (1:1,000; sc56-07, Novus Biologicals, Centennial, CO, USA), leukemia inhibitory
factor (LIF,1:1,000; sc-515931, Santa Cruz Biotechnology, Dallas, TX, USA), homeobox gene A10(HOXA10, 1:1,000; sc-271428, Santa Cruz
Biotechnology), and β-actin (1:10,000; 66009–1-Ig, Proteintech) overnight at 4°C. The membranes were then incubated with a horseradish peroxidase-conjugated goat anti-rabbit antibody
(1:5,000; AB0101, Abways, Shanghai, China) or goat anti-mouse antibody (1:10,000; A21010, Abbkine, Wuhan, China) at room temperature for 1 h. Protein bands were visualized using enhanced
chemiluminescence (ECL) and quantified with the ImageJ software (version 2.1; National Institutes of Health, Bethesda, MD, USA).
5
Protein Extraction and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To extract proteins, we treated cells or tissues with RIPA Lysis Buffer (Beyotime, China) which containing phenylmethanesulfonyl fluoride (PMSF) (Beyotime, China). The protein concentration was measured by the BCA kit (Beyotime, China) and 20ug of total protein was added to each well for protein separation by SDS-PAGE. Then the protein was transfered to PVDF membranes (Millipore, USA), blocked with 5% nonfat milk and incubated with primary and secondary antibodies for immunoblotting. Finally, the protein bands were detected by the ECL chemiluminescent substrate kit (Biosharp, china). The primary antibodies were as follows: PAFAH1B3 rabbit monoclonal (1:1000, ab166906, Abcam), E-cadherin rabbit monoclonal (1:10000, ab40772, Abcam), N-cadherin rabbit monoclonal (1:5000, ab76011, Abcam), SNAIL+SLUG rabbit monoclonal (1:1000, ab85936, Abcam). GAPDH mouse monoclonal (1:100000, 60,004–1-Ig, Proteintech). The secondary antibodies were: anti-rabbit secondary antibody (HRP) (1:5000, AB0101, Abways Technology), anti-mouse secondary antibody (HRP) (1:2000, AB0102, Abways Technology).
6
Western Blot Analysis of CX3CR1 and CX3CL1
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The cells and liver tissues were lysed with RIPA containing completed protease and phosphatase inhibitor, collected for centrifugation. The protein concentration was measured by a BCA Protein Assay Kit (Pierce Biotechnology, Waltham, MA). After denaturation with loading buffer, the cell lysates were separated with 10%-15% sodium dodecyl sulfate (SDS) –polyacrylamide gel electrophoresis (PAGE) gel electrophoresis and transferred to a nitrocellulose membrane. The membrane was blocked with 5% (m/v) skim milk and incubated overnight at 4°C with primary antibodies: CX3CR1(1 : 1000, Proteintech,13885-1-AP), CX3CL1 (1 : 1000, Proteintech, 10108-2-AP), and GAPDH (1 : 10000, Abways, AB2000). Then, the membrane was washed 3 times in TBST (10 min. each time) at room temperature and incubated with secondary antibodies: goat-anti-mouse (1 : 10000, Abways, AB0102) and goat-anti-rabbit (1 : 10000, Abways, AB0101) at room temperature for 1 hour. Finally, the membrane exposure was performed using enhanced chemiluminescence (ECL) by the Bio-Rad system.
7
Protein Expression Analysis in Cardiomyocytes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total protein was extracted by Whole-Cell Lysis Assay (KeyGEN BioTECH) from NRCMs and heart tissues, and the concentration was defined by the BCA Protein Assay Kit (KeyGEN BioTECH). Protein extractions were boiled and denatured, separated by 10% SDS–PAGE and then transferred onto PVDF membranes (Millipore). After blocking with 5% skimmed milk powder, the PVDF membranes were incubated overnight at 4 °C with the following rabbit-sourced primary antibodies, including anti-METTL3 antibody (Abcam, ab195352, 1:1000), anti-ANP antibody (Abcam, ab189921, 1:1000), anti-BNP antibody (Abcam, ab239510, 1:1000), anti-DKK2 antibody (Abcam, ab95274, 1:1000), anti-β-catenin antibody (Abcam, ab32572, 1:1000), anti-c-Myc antibody (Abcam, ab32072, 1:1000), and anti-β-actin antibody (ABways, AB0035, 1:5000). Subsequently, the membranes were washed with TBST and incubated with HRP-conjugated secondary antibody (Abways, AB0101, 1:5000; AB0102, 1:2000) at room temperature for 1 h. Finally, bands were detected using ECL (KeyGEN BioTECH) with a chemiluminescence system (Bio-Rad).
8
Protein Extraction and Western Blot Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein was extracted from mouse kidney tissues using RIPA lysis buffer (Solarbio, R0010). Equal amounts of protein were separated by 12.5% sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and then transferred to 0.45 μm polyvinylidene difluoride (PVDF) membranes. The membranes were blocked with 5% fat-free milk in Tris-buffered saline with Tween 20 (TBST) for 2 h at room temperature and then incubated overnight at 4°C with a primary antibody, followed by incubation with the HRP-conjugated secondary antibody (1:20000, AB0101, Abways) for 1 h at room temperature. The following primary antibodies were used: α-SMA (ab124964, Abcam, 1:30000), PYCARD (67824, CST, 1:1000), Glucocorticoid receptor (1:1000, CY6612, Abways), SIRT3 (10099-1-AP, Proteintech, 1:3000), and GAPDH (10494-1-AP, Proteintech, 1:30000). The protein blots were visualized with ECL.
9
Western Blot Analysis of STAT1 Pathway
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The following antibodies were used: GAPDH (1:5000; 5174S, CST, USA), PARP9 (1:250; ab53796 Abcam, USA), total STAT1 (1:1000, A19563, Abclonal, China), Phospho-STAT1-Y701 (1:1000, AP0054, Abclonal) and horseradish peroxidase-conjugated goat anti-rabbit secondary antibody (1:5000, AB0101, Abways). The relative densitometric analysis was conducted using the Image J.
10
Western Blot Analysis of Protein Expression
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were lysed with RIPA lysis buffer (Beyotime, P0013B) supplemented with 1 mM of the protease inhibitor phenylmethylsulfonyl fluoride (Beyotime, ST506) for 20 min on ice. After centrifugation at 12,000 rpm, the supernatant was collected, loaded onto a 10% polyacrylamide SDS gel, and subsequently transferred to a PVDF membrane (Millipore, IPVH00010). Membranes were blocked with TBST containing 5% skimmed milk (BD Pharmingen™, 232100) for 1 h at room temperature, followed by overnight incubation at 4°C with primary antibodies. After 5 washes with TBST, the membranes were incubated with secondary antibody at room temperature for 2 h. The membranes were then scanned and analyzed using the ChemiDoc™ system (Bio-Rad). The following antibodies were used: TGIF2 (11522-1-AP, Proteintech); GAPDH (EPR16891, Abcam); N-Cadherin (22018-1-AP, Proteintech); secondary antibody (AB0101, Abways).
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!