Ab125687

Manufactured by Abcam

Sourced in United States, United Kingdom

Ab125687 is a laboratory equipment product. It is designed for use in various scientific and research applications.

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Lab products found in correlation

5 protocols using ab125687

Quantitative Western Blot Analysis

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Western blot assay was used to detect total caspase-3, plasminogen activator inhibitor 1 (PAI1), protein kinase B (PKB) commonly known as Akt, p-Akt, 3-nitrotyrosine, and glyceraldehyde phosphate dehydrogenase (GAPDH). After electrophoresis and transfer, the polyvinylidene fluoride membranes were rinsed briefly in Tris-buffered saline, blocked (5% skim milk or 0.5% bovine serum albumin (BSA)) for 1 hr, and washed 3 times with Tris-buffered saline containing 0.05% Tween 20 for 5 min. The membranes were then incubated with anti-caspase-3 (Abcam, ab2171), anti-PAI1 (Abcam, ab125687), anti-p-AKT (Abcam, EP2109Y), anti-AKT (Abcam, ab8805), or anti-3-nitrotyrosine (Abcam, ab61392) in conjunction with GAPDH (Abcam, ab181602) overnight before being washed as described previously and then reacted with horseradish peroxidase-conjugated antibody for 1 hr. Antigen-antibody complexes were visualized by an electrochemiluminescence (ECL) kit (Biotrand, Crystal Lake, Ilinois, US) (12 (link)), and protein levels were normalized to GAPDH.

Liang Q., Wang B., Pang L., Wang Y., Zheng M., Wang Q., Yan J, & Xu J. (2016). Application of citrate as a tricarboxylic acid (TCA) cycle intermediate, prevents diabetic-induced heart damages in mice. Iranian Journal of Basic Medical Sciences, 19(1), 43-48.

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Molecular Mechanisms of PAI-1 Signaling

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Antibodies against PAI-1 (ab222754, 1:500; ab125687, 1:100), anti-VTN (ab45139, 1:2000), anti-LRP1 (ab168454, WB 1:1000, IP 1:100), anti-SMAD2 (phosphor S467) (ab280888), anti-SMAD2(ab40855), anti-SMAD3 (phosphor S423+S425) (ab172202), anti-SMAD3 (AB40854) were purchased from Abcam. Anti-PLAT (sc-515562, 1:50) and anti-PLAU (sc-59727, 1:500) were purchased from Santa cruz. Anti- Phospho-STAT1 (Tyr701) (7649T), anti-STAT1 (14994T), anti-Phospho-p44/42 MAPK (Erk1/2) (Thr202/tyr204) (4370), anti p44/42 MAPK (Erk1/2) (4695), anti-Phospho-Akt (Ser473) (4060), anti-Akt (14702) anti-phospho-PI3 Kinase p85 (Tyr458)/p55 (Tyr199) (4228), anti-PI3 Kinase p85(4257), anti-E-Cadherin (3195T), anti-N-Cadherin (13116T), anti-phospho- NF-κB p65 (Ser536) (3033s), anti- NF-κB p65 (8242) were purchased from CST. Anti-Anti-GAPDH (10494-1-AP) was purchased from Proteintech. His tagged human PAI-1 (10296-H08H) was purchased from Sino Biological. His-PLAU (PLU-H5229) was purchased from Acro. Human Lys-plasminogen (HPG2002), Glu-plasminogen (HPG2001) and S-2251 (100-01) were purchased from Enzyme Research Laboratories. Z-GGR-AMC was purchased from A+ peptide (shanghai). Recombinant Human LRP1-Cluster II-Fc Chimera protein (2368-L2) was purchased from Bio-Techne.

Zheng Y., Meng L., Qu L., Zhao C., Wang L., Liu C, & Shou C. (2023). Anti-PAI-1 Monoclonal Antibody Inhibits the Metastasis and Growth of Esophageal Squamous Cell Carcinoma. Journal of Cancer, 14(1), 114-128.

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Immunoblotting Protein Quantification Protocol

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Immunoblotting was done by a method as described previously [14 (link)]. Protein quantification was done by Bradford assay and equal quantity of protein was loaded onto each lane. The primary antibodies used in the study were mouse anti-α-SMA (ab7817, 1:1000, Abcam, Cambridge, MA, USA), anti-vimentin (D21H3, 1:1000, Cell Signalling Technology, Beverly, MA, USA) rabbit anti-fibronectin (AF5335, 1:1000, Affinity Biosciences, Brisbane, Queensland, Australia), mouse anti-fibulin-5 (ab66339, 1:800, Abcam, Cambridge, MA, USA), mouse anti-PAI-1 (ab125687, 1:1000, Abcam, Cambridge, MA, USA), rabbit anti-Fibrillin (AF0429, 1:1000, Affinity Biosciences, Brisbane, Queensland, Australia) and rabbit anti-pSMAD3(C25A9, 1:1000, Cell Signalling Technology, Beverly, MA, USA). Rabbit anti-GAPDH (1:10,000; Abcam, Cambridge, MA, USA) was used as the loading control.

Chakraborty M., Sahay P, & Rao A. (2021). Primary Human Trabecular Meshwork Model for Pseudoexfoliation. Cells, 10(12), 3448.

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STZ-Induced Diabetic Kidney Fibrosis

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Dulbecco's modified Eagle's medium (DMEM) and foetal bovine serum (FBS) were from Gibco (Grand Island, NY). Streptozotocin (STZ) was obtained from Sigma Chemical (St. Louis, MO). Primary antibodies for immunohistochemistry and Western blot analyses included PTPN2 (ab180764; Abcam); P‐STAT1 (#9167; Cell Signaling); P‐STAT3 (#9145; Cell Signaling); Arginase I (16001‐1‐AP; Proteintech Group Inc); Arginase II (14825‐1‐AP; Proteintech Group Inc); CD11c (17342‐1‐AP; Proteintech Group Inc); CD206 (18704‐1‐AP; Proteintech Group Inc); F4/80 (27044‐1‐AP; Proteintech Group Inc); CD3 (17617‐1‐AP; Proteintech Group Inc); fibronectin (ab2413; Abcam); Collagen I (#84336; Cell Signaling); Collagen IV (ab6586; Abcam); TGF‐β (#3711; Cell Signaling); PAI‐1 (ab125687; Abcam); α‐SMA (#19245; Cell Signaling); MCP‐1(#2029; Cell Signaling); TNF‐α (ab6671; Abcam); IL‐6 (ab208113; Abcam); ICAM‐1 (ab171123; Abcam); vascular endothelial growth factor (VEGF; 19003‐1‐AP; Proteintech Group Inc); CD31 (#77699; Cell Signaling) and β‐actin (ab8226; Abcam). ELISA kits for MCP‐1 were from ab208979, Abcam.

Li Y., Zhou H., Li Y., Han L., Song M., Chen F., Shang G., Wang D., Wang Z., Zhang W, & Zhong M. (2019). PTPN2 improved renal injury and fibrosis by suppressing STAT‐induced inflammation in early diabetic nephropathy. Journal of Cellular and Molecular Medicine, 23(6), 4179-4195.

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Vein Endothelial Cell Protein Analysis

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Vein endothelial cells were homogenized in lysis buffer containing protease inhibitor and centrifuged at 8,000 rpm at 4°C for 10 min. The supernatant of the mixture was used for analysis of the relevant protein using sodium dodecyl sulfate (SDS) assay according to the manufacturer's instructions (26 (link)). The primary goat anti-rat antibodies [anti-TAFI (1:1,000; ab181990), anti-PAI-1 (1:1,000; ab125687), anti-vWF (1:1,000; ab6994), anti-ADP (1:1,000; ab22554), anti-MMP9 (1:1,000; ab73734), anti-NF-κB (1:1,000; ab32360) (all from Abcam, Cambridge, UK)] were added after blocking (5% skimmed milk) for 60 min at 37°C and then washing with PBS three times. Subsequently, incubation with the secondary rabbit anti-goat antibody (1:1,000; ab6741; Abcam, UK) was carried out for 24 h at 4°C. The results were visualized using a chemiluminescence detection system.

Ma J., Li X., Wang Y., Yang Z, & Luo J. (2017). Rivaroxaban attenuates thrombosis by targeting the NF-κB signaling pathway in a rat model of deep venous thrombus. International Journal of Molecular Medicine, 40(6), 1869-1880.

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