Tissues from left atrium were used for western blot, and proteins including Nrf2 (1:1,000; Abcam), HO-1 (1:1,000; Abcam), Cav1.2 (1:1,000, Abcam), Kv1.5 (1:1000, Bioss), Kv4.2 (1:1000, Abcam), Kv4.3 (1:1000, Bioss), Cx40 (1:1000, Bioss), TH (1:1000, Abcam), GAP43 (1:1000, Abcam), TGF-β1 (1:1000, Abcam), Collagen I (1:1000, Bioss), Collagen III (1:1000, Bioss) and GAPDH (1:1000, Servicebio) were measured based on our previous study (Chen et al., 2020 (link)).
Collagen 3
Manufactured by Bioss Antibodies
Sourced in China
Collagen III is a type of collagen protein found in various connective tissues in the body. It plays a role in the structural integrity and function of these tissues. This lab equipment product can be used for research and analysis related to collagen III.
Automatically generated - may contain errors
Lab products found in correlation
Collagen 1, by Abcam (2 mentions)
Bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Gapdh, by Wuhan Servicebio Technology (1 mentions)
Collagen 1, by Bioss Antibodies (1 mentions)
Tgf β1, by Abcam (1 mentions)
Gap43, by Abcam (1 mentions)
P akt, by Cell Signaling Technology (1 mentions)
Gapdh, by Abcam (1 mentions)
Phosstop, by Roche (1 mentions)
Tubulin, by Abcam (1 mentions)
Calponin, by Abcam (1 mentions)
Elastin, by Bioss Antibodies (1 mentions)
Lamc1, by Cell Signaling Technology (1 mentions)
Goat anti rabbit cy3, by Proteintech (1 mentions)
Mmp 9, by Abcam (1 mentions)
Vimentin, by Abcam (1 mentions)
Quantity one software, by Bio-Rad (1 mentions)
Gapdh, by Proteintech (1 mentions)
3 protocols using collagen 3
1
Atrial Protein Expression Analysis
2
Protein Expression Analysis in Atrial Tissue
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Using RIPA lysis buffer with PhosSTOP (#04906845001, Roche, IN), the total proteins were extracted from the atrial tissue and quantified using the BCA Protein Assay Kit (Thermo Fisher Scientific, MA). Next, the samples (20 μg protein) were separated on the SDS-PAGE gel, and these proteins were transferred onto a PVDF membrane. After blocking with TBST buffer with 5% bovine serum albumin, the PVDF membranes were incubated overnight at 4°C with the following primary antibodies: tubulin (1 : 2000, Abcam, USA), GAPDH (1 : 5000, Abcam), collagen I (1 : 500, Abcam), collagen III (1 : 1000, Bioss, China), ɑ-SMA (1 : 1000, Abcam), PTEN (1 : 1000, Abcam), p-AKT (1 : 1000, Cell Signaling Technology, USA), PI3K (1 : 1000, Cell Signaling Technology), AKT (1 : 1000, Cell Signaling Technology), CACNA1C (1 : 1000, Bioss, China), KCNA5 (1 : 1000, Bioss, China), and CX43 (1 : 1000, Bioss, China). These membranes were washed thrice with 1× TBST buffer followed by incubation with HRP-labeled secondary antibodies (goat anti-rabbit and anti-mouse) for 1 h at room temperature. ImageJ was used to digitize and measure the optical intensities of the protein bands relative to those of tubulin or GAPDH.
3
Protein Expression Analysis of Stem Cells
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A radio-immunoprecipitation assay (RIPA) lysis buffer (Yisheng Biological Technology Co., Ltd., Shanghai, China) including proteinase suppressors was applied to isolate total protein. The concentration of total protein was detected by employing bicinchoninic acid (BCA) method (ComWin Biotech Co., Ltd., Beijing, China). A 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) was used to separate 20 µg of protein samples. Next, the samples were put onto a polyvinylidene difluoride (PVDF) membrane. After being blocked with 5% non-fat milk for 1 hour, the membrane was incubated with the antibodies overnight at 4 °C. The results were scanned by Quantity One software (version 4.6.9, Bio-Rad Laboratories, Hercules, CA, USA). The antibodies used in this part were as following, CD90 (Invitrogen Catalog #MA1-80650); CD44 (Invitrogen Catalog #MA5-17520); CD45 (Invitrogen Catalog #12-0461-82); Cav1 (Bioss,bs-1453R); MyHC (proteintech, 22281-1-AP); Goat anti-rabbit cy3 (proteintech SA00009-2); GAPDH (proteintech, 60004-1-Ig); MyoD (proteintech, 18943-1-AP); Collagen III (Bioss, bs-0549R); Collagen I (abcam, ab260043); LAMC1 (cell signaling, #92921); MMP1 (Affinify, DF6325); MMP9 (abcam, ab76003); HOXA11 (Bioss, bs-666R); Elastin (Bioss, bs-1756R); Calponin (abcam, ab46794); Vimentin (abcam, ab92547); GFP (abcam, ab1218).
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