SOCS5 (1:500, sc-100858, Santa Cruz); HA-Tag (1:1000, #3724, CST); hnRNP G (1:1000, ab190352, Abcam); FASN (1:1000, #3180, CST); Myc-Tag (1:1000, #2276, CST); ACC (1:1000, #3676, CST); SCD1 (1:1000, ab236868, Abcam); ACLY (1:5000, ab40793, Abcam); SREBP1 (1:1000, 14088-1-AP, Proteintech); IgG (1:1000, #2729, CST); HRP anti-mouse IgG (1:10,000, abs20001, Absin); HRP anti-rabbit IgG (1:10,000, abs20002, Absin); CD36 (1:1000, ab252922, Abcam); FATP4 (1:1000, ab200353, Abcam).
Abs20001
Manufactured by Absin
Sourced in China
The Abs20001 is a laboratory instrument designed to perform spectrophotometric analysis. It measures the absorbance or transmittance of light passing through a sample solution, providing data on the chemical composition and concentration of the sample.
Automatically generated - may contain errors
Lab products found in correlation
Abs20002, by Absin (6 mentions)
Abs132004, by Absin (2 mentions)
Ab236868, by Abcam (1 mentions)
Ab200353, by Abcam (1 mentions)
Ab190352, by Abcam (1 mentions)
Ab252922, by Abcam (1 mentions)
Ab40793, by Abcam (1 mentions)
Sc 100858, by Santa Cruz Biotechnology (1 mentions)
Ab225844, by Abcam (1 mentions)
Ab172967, by Abcam (1 mentions)
Ab19645, by Abcam (1 mentions)
Af5301, by Affinity Biosciences (1 mentions)
Clarity western ecl subs, by Bio-Rad (1 mentions)
Bax 50599 2 ig, by Proteintech (1 mentions)
Bca assay kit, by Solarbio (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Chemidoc imaging system, by Bio-Rad (1 mentions)
Gapdh, by Proteintech (1 mentions)
Precast glgel tris glycine page, by Sangon (1 mentions)
6 protocols using abs20001
1
Lipid Metabolism Protein Analysis
2
Antibody-based Cardiac Protein Analysis
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The antibodies were used as follows: Anti-ANP antibody (ab225844, abcam, WB:1:1000), Anti-BNP antibody (ab19645, abcam, WB:1:500), Anti-β-MHC antibody (ab172967, abcam, WB:1:1000), Anti-Ythdf2 antibody (24744–1-AP, proteintech, IF:1:200; WB:1:1000), Anti-CPT-1a (66039-1-Ig, proteintech, IF:1:200, WB:1:1000), Anti-PPARα (AF5301, Affinity, WB:1:1000) GAPDH antibody (abs132004, absin, WB:1:5000), anti-β-tubulin antibody (10094-1-AP, Proteintech, WB:1:1000), goat anti-rabbit IgG-HRP (abs20002, absin, WB:1:10000), goat anti-mouse IgG-HRP (abs20001, absin, WB:1:10000). AngII (HY-13948) was purchased from MCE (China).
3
Profiling Calpain and Apoptosis Markers in Immune Cells
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AFCs were lysed with RIPA lysis buffer (P0013B, Beyotime), and the protein concentration was determined by BCA assay kit (PC0020, Solarbio); 1/5 volume of loading buffer was added to the lysate, and then the sample was placed in boiling water for 10 min. Proteins are separated by sulfate–polyacrylamide gel electrophoresis (SDS‒PAGE) techniques and transferred to the PVDF membrane. PVDF membrane was blocked by 5% skim milk for 1 h, and then the membrane was incubated with primary antibodies at 4° overnight. Primary antibodies contain calpain1 (10,538–1-AP, Proteintech), calpain2 (11,472–1-AP, Proteintech), Bax (50,599–2-Ig, Proteintech), cleaved-Caspase 3 (9664, Cell Signaling Technology, MA, USA), Piezo1 (5939–1-AP, Proteintech), and Gapdh (60,004–1-Ig, Proteintech). PVDF membrane was washed by TBST and then was incubated with the secondary antibody for 1 h. The secondary antibodies contain Goat Anti-Mouse IgG (H + L) HRP (abs20001, Absin, Shanghai, China) or Goat Anti-Rabbit IgG (H + L) HRP (abs20002, Absin). The expression of the target protein was detected by Clarity Western ECL Subs (1705060SP, Bio-Rad, Universal Hood III, CA, USA) and ChemiDoc imaging system (Bio-Rad) and analyzed by ImageJ software. All results were quantified and normalized to GAPDH.
4
Integrated Omic and Protein Analysis
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Four cytokines (IL-1β, TNF-α, IL-17 and IL-6), four DEPs (BPGM, GPD1, APOE and APOC2) were identified by the integrated Omic analysis, and five key proteins (LRP5, DVL1, β-catenin, Cacybp, Skp1) involved in the Wnt/β-catenin signaling pathway were further validated by Western blot analysis. Briefly, total proteins were extracted using Cell lysis buffer (PMSF 1:100) for Western and IP (Beyotime, China). The total protein concentrations were quantified with a BCA assay (Bio-Rad, Hercules, CA). Next, the target proteins were separated by 8–20% Precast-Glgel Tris-Glycine PAGE (Sangon, Biotech), transferred to PVDF membrane. Then the PVDF membrane was blocked for 1 h. Next, the PVDF membrane was incubated with β-actin, IL-1β, TNF-α, IL-17, IL-6, BPGM, GPD1, APOE, APOC, LRP5, DVL1, β-catenin, GSK-3β, Cacybp and Skp1 overnight at 4 °C. Subsequently, the PVDF membrane was washed three times in TBST and incubated with either secondary antibody goat anti-mouse IgG-HRP (diluted 1: 2000, absin, abs20001, China) or goat anti-rabbit IgG-HRP (diluted 1: 2000, absin, abs20002, China) for 1 h. Protein bands were detected with the help of a FluorChem Q scanner (Proteinsimple, CA, USA).
5
Western Blot Analysis of Neurological Markers
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Equal amount of protein from the right hemisphere of the control or HI groups was separated on 4–12% SDS–PAGE gels (Beyotime) and transferred to polyvinyl difluoride membranes (Sigma Merck). The blots were then blocked with 5% bovine serum albumin in TBS buffer for 1 h at room temperature and probed overnight at 4°C by incubation with the following primary antibodies: Src (1:1,000, CST, catalog number: #2123), phosphor-Src (Tyr 416, 1:1,000, CST, catalog number: #6943), NMDAR subunit NR2B (1:1,000, CST, catalog number: #14544), phospho-NR2B (Tyr 1472, 1:1,000, CST, catalog number: #4208), myelin basic protein (MBP) (1:1,000, Biolegend, SMI99), Iba-1 (1:1,000, Abcam, ab178847), glial fibrillary acidic protein (GFAP) (1:1,000, CST, 12389), β-actin (1:5,000, Absin, abs137975), and GAPDH (1:1,000, Absin, abs132004), followed by appropriate secondary HRP-conjugated antibodies (anti-rabbit, 1:5,000, Absin, abs20002; anti-mouse, 1:5,000, Absin, abs20001). Blots were visualized with enhanced chemiluminescence reagents (BD Pharmingen) and Bio-Rad Image software.
6
Western Blot Analysis of Spleen Proteins
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Proteins were extracted from the spleen and quantified using the BCA method. Equal amounts of protein per sample were resolved with 10% SDS-PAGE and transferred to a 0.45-mm nitrocellulose membrane (EMD Millipore, Billerica, MA, USA) for 1 h. The membranes were blocked with 5% skim milk for 2 h at room temperature and then incubated overnight with the specific primary antibodies [diluted 1:1,000 in 5% skim milk in TBST (10 mM Tris-HCl, pH 8.0, 150 mM NaCl, 0.15% Tween-20)] against GRB2 (ab32037; Abcam), β-Actin (bsm-33036M; Bioss Bioscience), and IE2 (MAB8131; Millipore). The membranes were washed and separately placed in secondary antibodies. The membranes of GRB2 and β-actin were placed in anti-mouse IgG-HRP (abs20001; Absin Bioscience), and the membrane of IE2 was placed in anti-rabbit IgG-HRP (abs20002; Absin Bioscience) (both diluted 1:2,000 in 5% skim milk in TBST) for 2 h at room temperature. After membranes were washed three times with TBST at room temperature, positive bands were developed using the SuperSignal West Pico Trial kit (Thermo Fisher Scientific, Inc.) and detected with the VilberLourmat imaging system (VilberLourmat, Marne-la-Vallée, France). (Beckman Coulter). The results were analyzed with the software FlowJo Version 10 (TreeStar).
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