Recombinant mouse CXCL10 was obtained from PeproTech EC (London, UK). To detect specific protein expression by western blot, antibodies against P65 (cat. #8242), IκBα (cat. #4812) and α-tubulin (cat. #2144) were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against Lamin B (cat. #sc-6217) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA). The antibody against β-actin (cat. #A5441) was obtained from Sigma-Aldrich (St Louis, MO, USA). Secondary antibodies conjugated with horseradish peroxidase were purchased from Sigma-Aldrich.
Secondary antibodies conjugated with horseradish peroxidase
Manufactured by Merck Group
Sourced in United States
Secondary antibodies conjugated with horseradish peroxidase are laboratory reagents used for the detection and visualization of target proteins in various immunoassays, such as Western blotting and ELISA. These antibodies recognize and bind to the primary antibodies that are specific to the target proteins, and the conjugated horseradish peroxidase enzyme catalyzes a colorimetric or chemiluminescent reaction, allowing for the indirect detection of the target proteins.
Automatically generated - may contain errors
Lab products found in correlation
Recombinant mouse cxcl10, by Thermo Fisher Scientific (1 mentions)
Gata4, by Santa Cruz Biotechnology (1 mentions)
Beta actin actb, by Merck Group (1 mentions)
Connexin43 cx43, by Abcam (1 mentions)
Matrigel basement membrane matrix, by Corning (1 mentions)
P gsk3β s9, by Cell Signaling Technology (1 mentions)
Enhanced chemi luminescence (ecl), by GE Healthcare (1 mentions)
Antibodies against n cadherin, by Cell Signaling Technology (1 mentions)
Proliferating cell nuclear antigen (pcna), by Cell Signaling Technology (1 mentions)
Gsk3β, by Cell Signaling Technology (1 mentions)
E cadherin, by Cell Signaling Technology (1 mentions)
Laminin, by Abcam (1 mentions)
Wet system, by Bio-Rad (1 mentions)
Fibronectin, by Abcam (1 mentions)
β tubulin, by Abcam (1 mentions)
Type 1 collagen, by Abcam (1 mentions)
Chemidoc, by Bio-Rad (1 mentions)
Clarity western ecl substrate kit, by Bio-Rad (1 mentions)
4 protocols using secondary antibodies conjugated with horseradish peroxidase
1
Protein Expression Detection Protocol
2
Western Blotting Analysis of Cardiac Markers
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Western Blotting was performed as previously described [22] (link). Briefly, the protein samples were mixed with Laemmli sample buffer (1×) and boiled for 5 min. Proteins were subjected to 10% SDS-PAGE and electroblotted onto BioRad, 0.22 µM nitrocellulose membrane (BioRad Laboratories, USA), followed by blocking for 1 h at room temperature using Tris-buffered saline with 0.2% Tween 20 (TBS-T) containing 3% BSA. The membrane was incubated with primary antibodies at 4°C overnight and subsequently incubated with secondary antibodies conjugated with horseradish peroxidase (Sigma Aldrich, USA) for 2 h at RT and washed again with TBS-T. Antibody-reactive proteins were detected by enhanced chemiluminescence using Pierce ECL Plus western blotting detection reagents (Pierce, USA). Primary antibodies for GATA4 (1∶150; Santa Cruz Biotechnology, USA), Connexin43 (CX43; 1∶500; Abcam, USA), Alpha sarcomeric actin (ACTA1; 1∶500; Sigma Aldrich, USA) and Beta Actin (ACTB; 1∶1000; Sigma Aldrich, USA) were used along with goat anti-rabbit and anti-mouse horseradish peroxidase conjugated secondary antibodies (1∶10,000; Sigma Aldrich, USA).
3
Antibody-based Protein Expression Analysis
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All reagents were of analytical grade and were purchased from Sigma (St. Louis, MO, USA) unless otherwise specified. Cell culture dishes and flasks were from TPP (Switzerland). ECL was from GE Healthcare (Wauwatosa, WI, USA). Antibodies against PRDX6 were from Mybiosource (Cambridge, UK). Antibodies against N-cadherin, caspase-3, E-cadherin, PCNA, GSK3β, pGSK3βS9, and pAKTS473 were purchased from Cell Signaling (Danvers, MA, USA). Antibodies against AKT and actin were from Santa Cruz (Dallas, TX, USA). HA-flag antibody was from Thermo Fisher Scientific (Waltham, MA, USA), and complex I antibody and secondary antibodies conjugated with horseradish peroxidase were from Sigma. Secondary antibodies conjugated with Alexa Fluor 488 and 647 fluorophores and Alexa Fluor 488 phalloidin were from Abcam (Cambridge, UK), and Matrigel® Matrix basement membrane was from Corning (New York, NY, USA).
4
Western Blot Analysis of Extracellular Proteins
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Proteins were separated on a 12.5% SDS-PAGE gel and then transferred to a polyvinylidene difluoride membrane overnight at +4 °C at a constant voltage of 20 V using the Biorad wet system. The Toubin buffer system was used as the transfer buffer. Non-specific binding sites were blocked using 5% BSA in TBST (20 mM Tris-HCl, pH 7.6, 150 mM NaCl, and 0.1% Tween 20) for 1 h. After blocking, the membrane was incubated with primary antibodies specific for ALIX (Abcam, Cambridge, UK; dilution 1/1000), beta-tubulin (Abcam, Cambridge, UK; dilution 1/1000), fibronectin (Abcam, Cambridge, UK; dilution 1/2000), HSP70 (Biocat, Heidelberg, Germany; dilution 1/1000), CD63 (Merck Millipore, Burlington, MA, USA; dilution 1/1000), CD81 (BioLegend, San-Diego, CA, USA; dilution 1/500), collagen type I (Abcam, Cambridge, UK; dilution 1/1000), collagen type IV (Thermo Fischer Scientific, Waltham, MA, USA; dilution 1/1000), or laminin (Abcam, Cambridge, UK; dilution 1/2000) in the blocking solution overnight at +4 °C. The membrane was then washed with TBST and incubated with secondary antibodies conjugated with horseradish peroxidase (Sigma-Aldrich, St. Louis, MO, USA) in the blocking solution. Signal visualization was performed using the ClarityTM Western ECL Substrate kit (BioRad, Hercules, CA, USA) on ChemiDoc (BioRad, Hercules, CA, USA).
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