Chemiluminescence detection kit

Manufactured by PerkinElmer

Sourced in United States

The Chemiluminescence detection kit is a laboratory product designed for the detection and analysis of chemiluminescent signals. It provides the necessary reagents and materials to facilitate chemiluminescence-based experiments and assays.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Enhanced chemiluminescence detection kit (23 citations) Chemiluminescence kit (17 citations) Enzyme-linked chemiluminescence detection kit (6 citations) Western Lightning Plus-ECL chemiluminescence detection kit (4 citations) Enhanced chemiluminescence Western blot detection kit (3 citations) Western Lights chemiluminescence detection kit (3 citations) Western Lightning Plus-ECL enhanced chemiluminescence detection kit (3 citations) ECL chemiluminescence detection kit (3 citations) Enhanced chemiluminescence Western blotting detection kit (2 citations)

5 protocols using chemiluminescence detection kit

Protein Quantification and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Protein concentrations in the cell lysates were determined by Nanodrop ND-1000 (Gene Co., Ltd., Hong Kong, china). Once protein concentrations were determined, 50 µl of the protein specimens were denatured in 50 µl of 2×sample buffer (125 mmol/L Tris-HCl, pH 6.8, 20% glycerol, 10% β-mercaptoethanol, 0.02% bromophenol blue, and 4% SDS). Briefly, 30 µg of denatured protein were separated using a sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred to a polyvinyl difluoride (PVDF) membrane, which was blocked in 5% milk proteins that were suspended in tris-buffered saline tween-20 (TBST) for 2 h at room temperature. The membrane was rinsed three times with TBST followed by incubation with the primary antibody (Cell Signaling Technology, Inc, USA) overnight at 4°C. The membrane was then washed and treated with anti-rabbit secondary antibodies that were conjugated with horseradish peroxidase at a 1∶5000 dilution (Bioworld Technology Inc, USA). The immunoreactive proteins were visualized with a chemiluminescence detection kit (PerkinElmer, USA), and the expression of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) protein was used as the loading control across all samples analyzed by Western blot.

Yao Z., Song X., Cao S., Liang W., Lu W., Yang L., Zhang Z, & Wei L. (2014). Role of the Exogenous HCV Core Protein in the Interaction of Human Hepatocyte Proliferation and Macrophage Sub-Populations. PLoS ONE, 9(9), e108278.

+ Open protocol

+ Expand

Western Blot Analysis of ATII and RLE Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

After treatment, ATII and RLE cells were washed in ice-cold phosphate-buffered saline (PBS) and solubilized in cell lysis buffer (Cell Signaling). The lysates were cleared by centrifugation for 10 min at 14,000 × g. Protein concentrations were determined by the Bradford assay using a commercial dye reagent (Bio-Rad), and samples containing equal amounts of protein were separated by SDS-PAGE and transferred onto nitrocellulose membranes (GE Healthcare Life Sciences) by using a Trans-Blot Turbo (Bio-Rad) transfer system. The following commercially available antibodies and dilutions were used for Western blotting: mouse anti-Na,K-ATPase subunit α1 (clone 464.6; 1:10,000) was from EMD Millipore; rat anti-STIM1 (1:1000) was from Cell Signaling Technology. Primary antibodies were detected with horseradish peroxidase-conjugated secondary goat anti-mouse antibodies (1:10,000; Bio-Rad) or goat anti-rabbit antibodies (1:2,000; Cell Signaling Technology) by using a chemiluminescence detection kit (Perkin-Elmer Life Sciences). Quantification of protein levels was performed by densitometric scanning with ImageJ 1.29X (NIH).

Keller M.J., Lecuona E., Prakriya M., Cheng Y., Soberanes S., Scott Budinger G.R, & Sznajder J.I. (2014). Calcium Release-Activated Calcium (CRAC) Channels Mediate the β2-Adrenergic Regulation of Na,K-ATPase. FEBS letters, 588(24), 4686-4693.

+ Open protocol

+ Expand

Western Blot Quantification of ELAVL1 and GSK3β

Check if the same lab product or an alternative is used in the 5 most similar protocols

Protein concentration was quantified by Bradford assay (Bio-Rad, Hercules/CA USA), and proteins were resolved in 12% polyacrylamide gel. Thereafter, proteins were transferred to nitrocellulose membranes (Santa Cruz Biotechnology Dallas/TX, USA) using a semi-dry transfer apparatus (Bio-Rad, Munich, Germany). Incubation with ELAVL-1/HuR (HuR 3A2) antibody 1:5000 (sc-5261, Santa Cruz Biotechnology) respectively GSK3β antibody 1: 2000 (cell signaling #9315) were performed overnight at 4°C. Blots were developed with a chemiluminescence detection kit (Perkin Elmer Inc. Waltham/MA USA), as recommended by the manufacturer.

Hoffman O., Burns N., Vadász I., Eltzschig H.K., Edwards M.G, & Vohwinkel C.U. (2017). Detrimental ELAVL-1/HuR-dependent GSK3β mRNA stabilization impairs resolution in acute respiratory distress syndrome. PLoS ONE, 12(2), e0172116.

+ Open protocol

+ Expand

Immunoblot Analysis of Runx2 and Cell Cycle

Check if the same lab product or an alternative is used in the 5 most similar protocols

Runx2 and cell cycle markers were analyzed by immuno-blot analysis as described previously (Galindo et al., 2005 (link); Galindo et al., 2007 (link)). Briefly, equal amounts of total cellular protein collected in the presence of the proteasome inhibitor MG132 (Calbiochem, San Diego, CA, USA) and Complete® cocktail of protease inhibitor (Roche Diagnostics, Mannhein, Germany) were resolved in 10% SDS-PAGE and transferred to polyvinylidene difluoride membranes (Immobilon-P; Millipore, Billerica, MA.). Blots were incubated with a 1:2,000 dilution of each primary antibody for 1 h. Rabbit polyclonal antibodies (Cdk4, sc-260; cyclin A, sc-596), mouse monoclonal antibody (cyclin D1, sc-20044), and goat polyclonal antibody (actin, sc-1615) were acquired commercially (Santa Cruz Biotechnology, Inc.). Runx2-specific mouse monoclonal 8G5 antibody was obtained from MBL International (Woburn, MA). Membranes with bound primary antibodies were incubated with horseradish peroxidase-conjugated secondary antibodies (Santa Cruz Biotechnology, Inc.) for 1 h. Immuno-reactive protein bands were visualized on a film (BioMax, Kodak) using a chemiluminescence detection kit (PerkinElmer Life Sciences), and signal intensities were quantitated by densitometry. Each experiment was repeated at least three times.

Varela N., Aranguiz A., Lizama C., Sepulveda H., Antonelli M., Thaler R., Moreno R.D., Montecino M., Stein G.S., van Wijnen A.J, & Galindo M. (2015). Mitotic inheritance of mRNA facilitates translational activation of the osteogenic-lineage commitment factor Runx2 in progeny of osteoblastic cells. Journal of cellular physiology, 231(5), 1001-1014.

+ Open protocol

+ Expand

Western Blot Analysis of Lung Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were homogenized in lysis buffer (Cell Signaling Technology). Lung samples were homogenized on ice with cold lysis buffer in a 10-fold (w/w) excess of lysis buffer with pH of 7.6 (Roche) using a Polytron PT 10–35 homogenizer (Thermo Fisher Scientific). The protein concentration was quantified by the Bradford assay (Bio-Rad), and proteins were resolved on 7.5 or 12.5% polyacrylamide gels. Thereafter, proteins were transferred to nitrocellulose membranes (Bio-Rad) using a Trans-Blot Turbo transfer system (Bio-Rad). Incubation with specific antibodies was performed overnight at 4°C. Blots were developed with a chemiluminescence detection kit (PerkinElmer Life Sciences), as recommended by the manufacturer. The bands were quantified by densitometric scanning (ImageJ). Anti-β-tubulin (Santa Cruz Biotechnology) and total Mlc (Cell Signaling Technology) antibodies were used as loading controls.

Shigemura M., Lecuona E., Angulo M., Homma T., Rodríguez D.A., Gonzalez-Gonzalez F.J., Welch L.C., Amarelle L., Kim S.J., Kaminski N., Budinger G.R., Solway J, & Sznajder J.I. (2018). Hypercapnia increases airway smooth muscle contractility via caspase-7-mediated miR-133a-RhoA signaling. Science translational medicine, 10(457), eaat1662.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!