Protein levels were measured by western blot analysis. Cells were washed several times with PBS before collection and lysed with modified RIPA buffer. Cells were completely lysed after repeated vortexing and supernatants were acquired though centrifugation at 14,000 × g for 20 min. Proteins were resolved by sodium dodecyl sulfate–polyacrylamide gel (SDS-PAGE) and afterward transferred to a polyvinylidenedifluoride (PVDF) membrane (IPVH00010; Millipore, Boston, MA, USA) before incubating with primary antibodies (α-SA, Catalog No. ab72592, UK; cTnT, Catalog No. ab45932, UK; HIF-1α, Catalog No. GTX 127309, USA; apelin, Catalog No. ab125213, UK; APJ, Catalog No. LS-C149246, USA). The membranes were subjected to three 5-min washes with TBST and incubated with anti-IgG horseradish peroxidase-conjugated secondary antibody (Southern Biotech, Birmingham, AL, USA) for 60 min at room temperature. After extensive washing, bands were detected by enhanced chemiluminescence. The band intensities were quantified using image software (image J 2×, version 2.1.4.7).
Anti igg horseradish peroxidase conjugated secondary antibody
Manufactured by Southern Biotech
Sourced in United States
The Anti-IgG horseradish peroxidase-conjugated secondary antibody is a laboratory reagent used for the detection and quantification of immunoglobulin G (IgG) in various biological samples. The antibody is conjugated to the enzyme horseradish peroxidase, which catalyzes a colorimetric reaction that can be measured spectrophotometrically. This allows for the indirect detection and quantification of target IgG proteins in the sample.
Automatically generated - may contain errors
Lab products found in correlation
5 protocols using anti igg horseradish peroxidase conjugated secondary antibody
1
Western Blot Analysis of Protein Levels
2
Cytomegalovirus Seroprevalence Assessment via ELISA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
As described by Kilgour et al, CMV ELISA testing (University of Birmingham, Birmingham UK) was used to ascertain participant’s CMV status. Briefly, mock and viral lysate were used to coat 95 well plate overnight at 4 °C. Using plasma from 3 CMV positive donors, a standard was prepared and added to the plate in a 1 in 4 serial dilution, alongside 1ul of healthy donor samples. After 1 h incubation at room temperature, the plate was washed and anti-IgG horseradish peroxidase conjugated secondary antibody (Southern Biotech, Alabama, USA) added to each well and incubated for a further 1 h in the dark, RT. After repeat wash steps, 100 μL of TMB ELISA peroxidase substrate (Rockland Immunochemicals, Pennsylvania, USA) was added and incubated for 10 min in the dark, RT. To stop the reaction, 100 μL of 1 mM HCl was added, prior to reading on an ELISA plate reader at 450 nm [44 (link)].
3
Quantification of Protein Levels by Western Blot
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein levels were measured by western blot. Cells were washed several times with PBS before collection and lysed with modified RIPA buffer. Cells were completely lysed after repeated vortexing, and supernatants were acquired though centrifugation at 14,000 × g for 20 min. Proteins were resolved by sodium dodecyl sulfatepolyacrylamide gel (SDS-PAGE) and transferred to a polyvinylidenedifluoride (PVDF) membrane (IPVH00010, Millpore, Boston, USA) before incubation with the primary antibodies overnight at 4 °C. The membranes were subjected to three 5-min washes with TBST and incubated with anti-IgG horseradish peroxidase–conjugated secondary antibody (Southern biotech, Birmingham, USA) for 60 min at room temperature. After extensive washing, bands were detected by enhanced chemiluminescence. The band intensities were quantified by using image software (image J 2×, version 2.1.4.7).
4
Protein Expression Profiling of MSCs
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein levels of BCL2, BAX, tumor necrosis factor alpha (TNFα), TUSC-2, vascular endothelial growth factor (VEGF) alpha, platelet-derived growth factor (PDGF) beta and transforming growth factor beta-1 (TGFβ1) were measured by western blot. After 24 hours of reoxygenation, MSCs were washed several times with phosphate-buffered saline before collection and lysing with modified RIPA buffer. Cells were completely lysed after repeated vortexing and supernatants were acquired though centrifugation at 14,000 × g for 20 minutes. Proteins were then resolved by SDS-PAGE and subsequently transferred to a polyvinylidenedifluoride membrane (IPVH00010; Millipore, Boston, MA, USA) before incubating with primary antibodies overnight at 4°C. The membranes were subjected to three 5-minute washes with Tris-buffered saline–Tween and then incubated with anti-IgG horseradish peroxidase-conjugated secondary antibody (Southern Biotech, Birmingham, Alabama, USA) for 60 minutes at room temperature. After extensive washing, bands were detected by enhanced chemiluminescence. The band intensities were quantified using image software (image J 2×, version 2.1.4.7).
5
Western Blot for VEGFA Protein Quantification
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Protein levels were measured by western blot as reported previously [5 (link), 18 (link), 19 (link)]. After washing several times with PBS, cells were collected and lysed with modified RIPA buffer. Cells were completely lysed after repeated vortexing. Supernatants were obtained though centrifugation at 14,000×g for 20 min. Proteins were resolved by sodium dodecyl sulfate polyacrylamide gel (SDS-PAGE) and transferred to a polyvinylidenedifluoride (PVDF) membrane (IPVH00010; Millipore, Boston, MA, USA) before incubating with the primary anti-VEGFA rabbit polyclone antibody (Abcam, UK) overnight at 4 °C. The membranes were incubated with anti-IgG horseradish peroxidase-conjugated secondary antibody (Southern Biotech, Birmingham, AL, USA) for 60 min at room temperature following three 5-min washes with TBST. The bands were detected by enhanced chemiluminescence after extensive washing, and band intensities were quantified using image software (image J 2×, version 2.1.4.7) [5 (link), 18 (link), 19 (link)].
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!