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Supersignal substrates

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SuperSignal substrates are chemiluminescent detection reagents designed for quantitative and qualitative analysis of proteins in Western blotting applications. They generate a luminescent signal upon reaction with the horseradish peroxidase (HRP) enzyme, which can be detected using a luminometer or X-ray film.

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Lab products found in correlation

Gorab, by Proteintech (2 mentions) Hrp conjugated secondaryantibodies, by BD (2 mentions) Polyvinylidene fluoride (pvdf), by Merck Group (2 mentions) β actin, by Santa Cruz Biotechnology (2 mentions)

3 protocols using supersignal substrates

1

Western Blot Quantification of Protein Targets

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protein was extracted in cold RIPA lysis buffer (50 mM Tris-HCl pH 7.4,
150 mM NaCl, 1% Triton X-100, 1% sodium deoxycholate, and
0.1% SDS) supplemented with proteinase inhibitors. Tissue or cell
lysates were cleared and separated by 10% SDS-PAGE and transferred to
polyvinylidene fluoride (PVDF, Millipore) or Hybond nitrocellulose (GE
Healthcare) membranes, following standard procedures. Blots were probed with the
primary antibodies which were then detected with HRP-conjugated secondary
antibodies (BD biosciences) and SuperSignal substrates (Thermo Scientific).
Enhanced chemiluminescent (ECL) substrate (Pierce, Rockford, IL USA) and
CL-XPosure film (Thermo Scientific) were used for detection. The following
primary antibodies were used: GORAB, 1:1,000 (Proteintech); β-actin,
1:1,000 (Santa Cruz); GLI1, 1:250 (clone V812, Cell Signaling); GLI3 (1
µg/ml, Cell Signaling). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH)
or β-actin was used as a loading control. Quantification was performed
with densitometry and ImageJ software (1.43u, NIH).
Liu Y., Snedecor E.R., Choi Y.J., Yang N., Zhang X., Xu Y., Han Y., Jones E.C., Shroyer K.R., Clark R.A., Zhang L., Qin C, & Chen J. (2015). Gorab is required for dermal condensate cells to respond to hedgehog signals during hair follicle morphogenesis. The Journal of investigative dermatology, 136(2), 378-386.
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2

Western Blot Analysis of Phospho-IRF3

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Cells were lysed in RIPA lysis buffer supplemented with 1x Halt Protease and Phosphatase Inhibitor Cocktail. Protein samples were separated by SDS-PAGE and then transferred to nitrocellulose membrane. Primary antibodies specific for phospho-IRF3 (1:1000, CST, 4947), IRF3 (1:1000, CST, 4302), IRF3 (1:1000, Millipore, MABF268), C7 antibody (1:500), FLAG (1:1000, Sigma, F3165), GAPDH (1:2000, CST, 2118) and β-actin (1:2000, CST, 4967) were use. β-actin or GAPDH were used as loading controls. Anti-rabbit or mouse HRP-linked IgG antibody was used as a secondary antibody (1:5000, CST, 7074 or 7076). Detection was performed using SuperSignal Substrates (Thermo Fisher, 34577 or 34095).
Yang N., Luna J.M., Dai P., Wang Y., Rice C.M, & Deng L. (2022). Lung type II alveolar epithelial cells collaborate with CCR2+ inflammatory monocytes in host defense against poxvirus infection. Nature Communications, 13, 1671.
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3

Western Blot Quantification of Protein Targets

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protein was extracted in cold RIPA lysis buffer (50 mM Tris-HCl pH 7.4,
150 mM NaCl, 1% Triton X-100, 1% sodium deoxycholate, and
0.1% SDS) supplemented with proteinase inhibitors. Tissue or cell
lysates were cleared and separated by 10% SDS-PAGE and transferred to
polyvinylidene fluoride (PVDF, Millipore) or Hybond nitrocellulose (GE
Healthcare) membranes, following standard procedures. Blots were probed with the
primary antibodies which were then detected with HRP-conjugated secondary
antibodies (BD biosciences) and SuperSignal substrates (Thermo Scientific).
Enhanced chemiluminescent (ECL) substrate (Pierce, Rockford, IL USA) and
CL-XPosure film (Thermo Scientific) were used for detection. The following
primary antibodies were used: GORAB, 1:1,000 (Proteintech); β-actin,
1:1,000 (Santa Cruz); GLI1, 1:250 (clone V812, Cell Signaling); GLI3 (1
µg/ml, Cell Signaling). Glyceraldehyde-3-phosphate dehydrogenase (GAPDH)
or β-actin was used as a loading control. Quantification was performed
with densitometry and ImageJ software (1.43u, NIH).
Liu Y., Snedecor E.R., Choi Y.J., Yang N., Zhang X., Xu Y., Han Y., Jones E.C., Shroyer K.R., Clark R.A., Zhang L., Qin C, & Chen J. (2015). Gorab is required for dermal condensate cells to respond to hedgehog signals during hair follicle morphogenesis. The Journal of investigative dermatology, 136(2), 378-386.
+ Open protocol
+ Expand

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