Supersignaltm west pico femto chemiluminescent substrate kit

Manufactured by Thermo Fisher Scientific

The SuperSignal West Pico/Femto Chemiluminescent Substrate kit is a lab equipment product designed for the detection and quantification of proteins in Western blot analysis. The kit provides a chemiluminescent substrate that reacts with the enzyme-labeled secondary antibody, producing a luminescent signal that can be captured and measured using a compatible imaging system.

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Lab products found in correlation

Bca kit, by Thermo Fisher Scientific (6 mentions) Phenyl methyl sulfonyl fluoride (pmsf), by Solarbio (3 mentions) Pvdf membrane, by Merck Group (3 mentions) Ab153904, by Abcam (1 mentions) Ab51072, by Abcam (1 mentions) Ab181602, by Abcam (1 mentions) Ripa lysis buffer, by Solarbio (1 mentions) Pa5 99584, by Thermo Fisher Scientific (1 mentions) Amersham imager 600, by Thermo Fisher Scientific (1 mentions) Ab52640, by Abcam (1 mentions)

3 protocols using supersignaltm west pico femto chemiluminescent substrate kit

Western Blot Analysis of Protein Markers

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Proteins were extracted using RIPA lysis buffer (Solarbio, R0020) containing PMSF (Solarbio, P0100), phosphatase inhibitor cocktail I and II (MCE, HY-K0021 and HY-K0022). In total, lysate containing 10–20 μg of protein was separated on SDS-PAGE and transferred to PVDF membranes (Millipore). Antibodies against MMP13 (ab51072), HOXC10 (ab153904) and GAPDH (ab181602) were from Abcam while antibodies against YEATS2 (24717-1-AP) and NECAB2 (12257-1-AP) were from Proteintech. The signal was captured with a SuperSignal^TM West Pico/Femto Chemiluminescent Substrate kit (Thermo Fisher, 34580) analyzedthrough the Amersham Imager 600.

Xi Y., Lin Y., Guo W., Wang X., Zhao H., Miao C., Liu W., Liu Y., Liu T., Luo Y., Fan W., Lin A., Chen Y., Sun Y., Ma Y., Niu X., Zhong C., Tan W., Zhou M., Su J., Wu C, & Lin D. (2022). Multi-omic characterization of genome-wide abnormal DNA methylation reveals diagnostic and prognostic markers for esophageal squamous-cell carcinoma. Signal Transduction and Targeted Therapy, 7, 53.

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Protein Extraction and Western Blot Analysis

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Cells were lysed with RIPA lysis buffer (Solarbio, R0020) containing PMSF (Solarbio, P0100), phosphatase inhibitor cocktail I (MCE, HY-K0021) and phosphatase inhibitor cocktail II (MCE, HYK0022). Total protein levels in cell lysates were determined by the BCA kit (Thermo Fisher Scientific). Lysates containing 10-20 μg of protein were subjected to SDS-PAGE separation and transferred to the PVDF membrane (Millipore) for determination. The antibody used for A3A (PA5-99584) was from Thermo Fisher. Vinculin (ab219649), STAT1 (ab234400), γH2AX (phospho S139, ab26350) antibodies were from Abcam, while antibodies against cGAS (#79978), STING (#13647), phospho-STING (p-STING) (Ser366; #50907), IRF3 (#11904), phospho-IRF3 (p-IRF3) (Ser396; #29047), TBK1 (#3504), phospho-TBK1 (p-TBK1) (Ser172; #5483), and phospho-STAT1 (p-STAT1) (Tyr701; #7649) were from Cell Signaling Technology. Antibodies against for PD-L1 (66248-1-Ig) was from Proteintech. All the antibodies were diluted in Universal Antibody Diluent (NCM Biotech, WB500D). The membrane was incubated overnight at 4°C with primary antibody and 2 hours with secondary antibody. The signal was detected with a SuperSignalTM West Pico/Femto Chemiluminescent Substrate kit (Thermo Fisher, 34580) through the Amersham Imager 600.

Yang J., Xiang T., Zhu S., Lao Y., Wang Y., Liu T., Li K., Ma Y., Zhong C., Zhang S., Tan W., Lin D, & Wu C. (2023). Comprehensive Analyses Reveal Effects on Tumor Immune Infiltration and Immunotherapy Response of APOBEC Mutagenesis and Its Molecular Mechanisms in Esophageal Squamous Cell Carcinoma. International Journal of Biological Sciences, 19(8), 2551-2571.

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Protein Expression Analysis with Inhibitors

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Cells were lysed with RIPA lysis buffer (Solarbio, R0020) containing PMSF (Solarbio, P0100), phosphatase inhibitor cocktail I (MCE, HY-K0021) and phosphatase inhibitor cocktail II (MCE, HY-K0022). The protein concentration of each sample was determined by BCA assay using the BCA kit (Thermo Fisher Scientific). Lysate containing 10–20 µg of protein was separated on SDS-PAGE and transferred to PVDF membranes (Millipore). Antibody against VAV2 (ab52640), Ku70 (ab92450), STAT1 (ab234400), phospho-STAT1 (S727) (ab109461), ATF7 (ab183507), GTF2E1 (ab140634), GAPDH (ab181602), HIS (ab213204), DNA PKcs (ab32566) or DNA PKcs (phosphor S2056) (ab124918) was from Abcam while antibodies against γ-H2AX (Ser139) (#2577), Ku80 (#2180), FLAG (#14793) or MYC (#2278) antibody was from Cell Signaling Technology. The signal was detected with a SuperSignal^TM West Pico/Femto Chemiluminescent Substrate kit (Thermo Fisher, 34580) through the Amersham Imager 600. The protein bands were quantified by gray scanning using ImageJ software.

Liu W., Miao C., Zhang S., Liu Y., Niu X., Xi Y., Guo W., Chu J., Lin A., Liu H., Yang X., Chen X., Zhong C., Ma Y., Wang Y., Zhu S., Liu S., Tan W., Lin D, & Wu C. (2021). VAV2 is required for DNA repair and implicated in cancer radiotherapy resistance. Signal Transduction and Targeted Therapy, 6, 322.

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