Thbs1

Manufactured by Santa Cruz Biotechnology

Sourced in United States

THBS1 is a lab equipment product manufactured by Santa Cruz Biotechnology. It is a protein that plays a role in various cellular processes, including cell-to-cell and cell-to-matrix interactions, angiogenesis, and wound healing.

Automatically generated - may contain errors

Lab products found in correlation

α sma, by Santa Cruz Biotechnology (1 mentions) β actin, by Santa Cruz Biotechnology (1 mentions) Protein assay kit, by Beyotime (1 mentions) Mmp 1, by Santa Cruz Biotechnology (1 mentions) Mini protean tgx stain free protein gel, by Bio-Rad (1 mentions) Timp 2, by Abcam (1 mentions) Goat anti mouse hrp, by Merck Group (1 mentions) Criterion stain free imaging system, by Bio-Rad (1 mentions) Serpine1, by Santa Cruz Biotechnology (1 mentions) Col6a1, by Santa Cruz Biotechnology (1 mentions) Ultrascence pico ultra western substrate, by GeneDireX (1 mentions) Goat anti rabbit hrp, by Merck Group (1 mentions) Chemidoc mp imaging system, by Bio-Rad (1 mentions) Trans blot turbo system, by Bio-Rad (1 mentions) Jetsi, by Polyplus Transfection (1 mentions) Imagequant 5, by GE Healthcare (1 mentions) Ponceau red, by Merck Group (1 mentions) Immobilon p, by Merck Group (1 mentions) Enhanced chemiluminescence ecl detection reagents, by GE Healthcare (1 mentions) Las 3000 image analyzer, by GE Healthcare (1 mentions)

6 protocols using thbs1

Investigating Fibrosis Markers in Cells

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Antibodies against CD47 and γ-catenin were obtained from Abcam (Cambridge, UK), while those against Col-1, α-SMA, THBS1, and β-actin were procured from Santa Cruz Biotechnology (Dallas, TX, USA). Rabbit anti-E-cadherin was purchased from Bioss Biotechnology (Beijing, China) and antibodies against vimentin and CD68 were supplied by MXB Biotechnologies (Fuzhou, China). Lipofectamine 2000 was purchased from Science Biotechnology (Invitrogen, Beijing, China) and the Protein Assay Kit was purchased from Beyotime Institute of Biotechnology (Jiangsu, China). Masson's trichrome (Masson) and Van Gieson (VG) staining kits were procured from Zhuhai Besso Biotechnology Institute (Wuhan, China). Leucine-serine-lysine-leucine (LSKL), a competitive TGF-β1 antagonist and an inhibitor of thrombospondin, was procured from MedChemExpress (MCE, Shanghai, China). Kits for the triglyceride (TG) assay, total cholesterol (TC) assay, LDL cholesterol assay, high-density lipoprotein cholesterol assay, and blood urea nitrogen (BUN) assay were purchased from Nanjing Jiancheng Bioengineering Institute.

Gao L., Yang T.T., Zhang J.S., Liu H.X., Cai D.C., Wang L.T., Wang J., Li X.W., Gao K., Zhang S.Y., Cao Y.J., Ji X.X., Yang M.M., Han B., Wang S., He L., Nie X.Y., Liu D.M., Meng G, & He C.Y. (2021). THBS1/CD47 Modulates the Interaction of γ-Catenin With E-Cadherin and Participates in Epithelial–Mesenchymal Transformation in Lipid Nephrotoxicity. Frontiers in Cell and Developmental Biology, 8, 601521.

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Protein Expression Analysis Workflow

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CM were mixed with 4× Laemmli sample buffer (containing 2-mercaptoethanol) and boiled for 10 min at 95°C. 15-μl protein volumes were separated on 4–15% Mini-PROTEAN TGX stain-free protein gels (4568086; Bio-Rad), and whole proteins were detected with the Criterion Stain-free imaging system (Bio-Rad). Next, the proteins were transferred on polyvinylidene difluoride membranes using the Trans-Blot-Turbo system (Bio-Rad). The membranes were blocked in 5% nonfat dry milk/TBS-T for 1 h at RT. The following primary antibodies were diluted 1:1,000 in 5% BSA/TBS-T and incubated overnight at 4°C: MMP1 (sc-30069; Santa Cruz), SERPINE1 (sc-5297; Santa Cruz), TIMP2 (ab53730; Abcam), COL6A1 (sc-377143; Santa Cruz), FN1 (F3648; Sigma-Aldrich), and THBS1 (sc-59887; Santa Cruz). Membranes were 3× washed in TBS-T and the following secondary antibodies were diluted 1:20,000 in 2.5% nonfat dry milk/TBS-T and incubated for 1 h at RT: goat-anti-mouse-HRP (SAB3701073-2; Sigma-Aldrich) or goat-anti-rabbit-HRP (SAB3700878-1; Sigma-Aldrich). HRP was visualized by the UltraScence Pico Ultra Western Substrate (CCH345-B; GeneDireX) and the ChemiDoc MP imaging system (Bio-Rad).

Blache U., Horton E.R., Xia T., Schoof E.M., Blicher L.H., Schönenberger A., Snedeker J.G., Martin I., Erler J.T, & Ehrbar M. (2019). Mesenchymal stromal cell activation by breast cancer secretomes in bioengineered 3D microenvironments. Life Science Alliance, 2(3), e201900304.

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Targeted siRNA Delivery for Nerve Injury

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Mice were deeply sedated with isoflurane and stereotactically injected with 1.2 μl of an 8-µM siRNA solution using JetSi (Polyplus Transfection) within the left S1 cortex hindlimb region (0.5 mm posterior to the bregma, 1.5 mm lateral to the midline, 0.15 mm from the pial surface). siRNA solutions were prepared according to the manufacturer’s protocols. All siRNAs were purchased from Santa Cruz Biotechnology (control siRNA, sc 37007; Thbs1, sc-3666; Gpc4 siRNA, sc-145457; Sparcl1 siRNA, sc-153239). siRNA injection experiments were performed immediately before the sciatic nerve ligation surgery.

Danjo Y., Shigetomi E., Hirayama Y.J., Kobayashi K., Ishikawa T., Fukazawa Y., Shibata K., Takanashi K., Parajuli B., Shinozaki Y., Kim S.K., Nabekura J, & Koizumi S. (2022). Transient astrocytic mGluR5 expression drives synaptic plasticity and subsequent chronic pain in mice. The Journal of Experimental Medicine, 219(4), e20210989.

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Quantitative Western Blot Analysis of THBS1 and TSP1

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Western blot analyses were performed utilizing standard procedures. Chondrocytes from 3 OA donors different from those selected for the proteomics strategy were used. Briefly, 20 μg of secreted proteins were loaded and resolved using 8% SDS-polyacrylamide gel electrophoresis (SDS-PAGE). The separated proteins were then transferred to polyvinylidene fluoride (PVDF) membranes (Immobilon P, Millipore Co., Bedford, MA, USA) by electro-blotting and probed with specific antibodies against THBS1 or TSP1 (Santa Cruz). Immunoreactive bands were detected by chemiluminescence using corresponding horseradish peroxidase (HRP)-conjugated secondary antibodies and enhanced chemiluminescence (ECL) detection reagents (GE Healthcare), then digitized using the LAS 3000 image analyzer. Equivalent loadings were verified by Ponceau Red (Sigma) staining after transference (data not shown). Quantitative changes in band intensities were evaluated using ImageQuant 5.2 software (GE Healthcare).

Calamia V., Mateos J., Fernández-Puente P., Lourido L., Rocha B., Fernández-Costa C., Montell E., Vergés J., Ruiz-Romero C, & Blanco F.J. (2014). A pharmacoproteomic study confirms the synergistic effect of chondroitin sulfate and glucosamine. Scientific Reports, 4, 5069.

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Western Blot Analysis of Protein Targets

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Proteins were extracted from the untreated, mock, and lenti-miR-1290 groups. Bradford assay was used to normalize the concentration of the extracted samples. 20 μg of each protein sample was subjected to SDS–PAGE in 14% acrylamide gels and transferred to a polyvinylidene difluoride membrane (Millipore, Bedford, MA, USA). A combination of 5% non-fat dry milk and 0.05% Tween-20 was used for blocking and washing in TBS and TBST, respectively. The blots were probed with primary antibodies, including and GAPDH, THBS1, DKK3, and SCAI (Santa Cruz, USA), which were diluted in TBST (1:1000) and incubated for 2 h at room temperature. The membranes were incubated with goat anti-mouse HRP-conjugated secondary IgG for 1 hour at room temperature. Afterward, the immunoreactive bands were detected using an ECL western blotting substrate (ECL, Amersham, Bucking Hampshire, UK).

Soheilifar M.H., Pornour M., Saidijam M., Najafi R., Azizi Jalilian F., Keshmiri Neghab H, & Amini R. (2021). miR-1290 contributes to oncogenesis and angiogenesis via targeting of THBS1, DKK3 and, SCAI. BioImpacts : BI, 12(4), 349-358.

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Western Blot Analysis of Exosomal Markers

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Samples in different groups were lysed by the RIPA buffer containing 1% protease inhibitor PMSF. The lysates were centrifuged and then supernatants were collected. Equal proteins were added on an 8–12% Bis–Tris gel and then transferred to a polyvinylidene difluoride membrane. After transfer, membrane was treated with blocking solution for 2 h and probed with primary antibody against P53 (Abcam, USA), CD9 (Abcam, USA), CD63 (Abcam, USA), CD81 (Abcam, USA), HSP70 (Abcam, USA), Ang-2 (CST, USA), VEGF (Novus, USA), THBS1 (Santa Cruz, USA), STAT1 (Novus, USA), and LIF (Abcam, USA) overnight at 4 °C, followed by horseradish peroxidase-conjugated secondary antibody for 1 h. The loading control was the constitutively expressed protein β-actin (Sigma, USA) or lamin B (Abcam, USA). The blots were visualized with enhanced chemiluminescence system and quantitated using ImageJ.

Chen L., Han Y., Li Y., Chen B., Bai X., Belguise K., Wang X., Chen Y., Yi B, & Lu K. (2019). Hepatocyte-derived exosomal MiR-194 activates PMVECs and promotes angiogenesis in hepatopulmonary syndrome. Cell Death & Disease, 10(11), 853.

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