The reference standard of glucose and uronic acid were provided by Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China). Moreover, metformin hydrochloride tablets were purchased from Sino-American Shanghai Squibb Pharmaceuticals Ltd. (Shanghai, China). STZ was obtained from Sigma-Aldrich (Sacramento, CA, USA). All test assay kits and enzyme-linked immunosorbent assay (ELISA) kits were supplied by NanJing JianCheng Bioengineering Institute (Nanjing, China). Tumor necrosis factor-alpha (TNF-α) was obtained from PeproTech Inc. (Cranbury, NJ, USA). Lipopolysaccharides (LPS) were provided by Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). NF-κB p65 (1:2,000), phospho-NF-κB p65(Ser536; 1:2,000), and phospho-IκBα (Ser32; 1:2,000) were purchased from Cell Signaling Technology (Beverly, MA, USA). The radioimmunoprecipitation assay (RIPA) lysis buffer with protease/phosphatase inhibitor cocktail, bicinchoninic acid (BCA) protein assay, anti-β-actin antibodies (1:2,000), and horseradish peroxidase (HRP)-conjugated secondary antibodies was purchased from Beijing ComWin Biotech Co., Ltd. (Beijing, China). All other chemical reagents were of analytical grade.
Horseradish peroxidase hrp conjugated secondary antibody
Manufactured by CoWin Biotech
Sourced in China
Horseradish peroxidase (HRP)-conjugated secondary antibodies are a type of immunoassay reagent. They are used to detect and quantify the presence of specific target proteins in biological samples through enzyme-linked immunosorbent assay (ELISA) techniques.
Automatically generated - may contain errors
Lab products found in correlation
Tumor necrosis factor alpha tnf α, by Thermo Fisher Scientific (2 mentions)
Phospho nf κb p65 ser536, by Cell Signaling Technology (2 mentions)
Phospho iκbα ser32, by Cell Signaling Technology (2 mentions)
Uronic acid, by Yuanye Bio-Technology (2 mentions)
Bicinchoninic acid bca protein assay, by CoWin Biotech (2 mentions)
Anti β actin antibodies, by CoWin Biotech (2 mentions)
Protease phosphatase inhibitor cocktail, by CoWin Biotech (2 mentions)
Lipopolysaccharides (lps), by Solarbio (2 mentions)
Streptozotocin (stz), by Merck Group (2 mentions)
Nf κb p65, by Cell Signaling Technology (2 mentions)
Cpt1a, by Santa Cruz Biotechnology (1 mentions)
Srebf1, by Abcam (1 mentions)
β actin, by CoWin Biotech (1 mentions)
Hmgcr, by Abcam (1 mentions)
4 protocols using horseradish peroxidase hrp conjugated secondary antibody
1
Antidiabetic and Anti-inflammatory Effects
2
Metformin Attenuates Inflammatory Response
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The reference standard of glucose and uronic acid were provided by Shanghai Yuanye Biotechnology Co., Ltd. (Shanghai, China). Moreover, metformin hydrochloride tablets were purchased from Sino-American Shanghai Squibb Pharmaceuticals Ltd. (Shanghai, China). STZ was obtained from Sigma-Aldrich (Sacramento, CA, USA). All test assay kits and enzyme-linked immunosorbent assay (ELISA) kits were supplied by NanJing JianCheng Bioengineering Institute (Nanjing, China). Tumor necrosis factor-alpha (TNF-α) was obtained from PeproTech Inc. (Cranbury, NJ, USA). Lipopolysaccharides (LPS) were provided by Beijing Solarbio Science & Technology Co., Ltd. (Beijing, China). NF-κB p65 (1:2,000), phospho-NF-κB p65(Ser536; 1:2,000), and phospho-IκBα (Ser32; 1:2,000) were purchased from Cell Signaling Technology (Beverly, MA, USA). The radioimmunoprecipitation assay (RIPA) lysis buffer with protease/phosphatase inhibitor cocktail, bicinchoninic acid (BCA) protein assay, anti-β-actin antibodies (1:2,000), and horseradish peroxidase (HRP)-conjugated secondary antibodies was purchased from Beijing ComWin Biotech Co., Ltd. (Beijing, China). All other chemical reagents were of analytical grade.
3
Immunohistochemical Analysis of E-cadherin and Vimentin
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The sections of lung tissues were deparaffinized in xylene and rehydrated in graduated ethanol solutions. Following microwave-based antigen retrieval with citric acid pretreatment, the sections were incubated in 1% hydrogen peroxide for 15 min to block endogenous peroxidase. Subsequently, the specimens were incubated with a mouse polyclonal antibody to E-cadherin (1:200) or a rabbit polyclonal antibody to vimentin (1:200) at 4°C overnight, respectively. The sections were then incubated with anti-mouse (cat. no. CW01025) or anti-rabbit (cat. no. CW01035) horseradish peroxidase (HRP)-conjugated secondary antibody (1:100; Beijing Com Win Biotech Co., Ltd.) for 30 min at room temperature, followed by staining with DAB (22 (link)). For the negative control, the primary antibody was replaced with PBS. The sections were observed under a light microscope (magnification, ×400). E-cadherin was mainly expressed in the cell membrane and cytoplasm, whereas vimentin was mainly expressed in the cytoplasm. The brown staining of a cell membrane or cytoplasm is a positive signal in protein immunohistochemistry. The mean integrated OD (IOD) was then detected using the Image Pro Plus 6.0 image analysis system for all the sections. The IOD value represented the expression of each protein. The mean IOD values of each group were compared.
4
Western Blot Analysis of Cholesterol Metabolism
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Protein samples were prepared from the cells according to the manufacturer’s instructions (ComWin Biotech, China) and quantified using the bicinchoninic acid (BCA) method. Ten micrograms of each protein sample was separated by SDS-polyacrylamide gel electrophoresis and then transferred onto CN membranes. Membranes were subsequently blocked with 5% skim milk and incubated with primary antibodies against SREBF1 (1:4000; Abcam, UK), LSS (1:4000; Abcam), HMGCR (1:4000; Abcam), FDFT1 (1:1000; Santa Cruz Biotech, Santa Cruz, CA), CPT1a (1:1000; Santa Cruz Biotech), SCARB2 (1:1000; Santa Cruz Biotech), and β-actin (1:4000, Beijing ComWin Biotech Co., Ltd.) overnight at 4°C. The membranes were then washed three times with TBS-T buffer and incubated with a horseradish peroxidase (HRP)-conjugated secondary antibody (1:4,000, Beijing ComWin Biotech Co., Ltd.) for 1 h at room temperature. The membranes were washed three times, followed by immunodetection using an enhanced chemiluminescence kit (Beijing ComWin Biotech Co., Ltd.).
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