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Chemiluminescent

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Chemiluminescent is a type of lab equipment that generates light through a chemical reaction. It is used to detect and measure the presence of specific molecules or substances in a sample.

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Lab products found in correlation

G box ichemi xt imager, by Syngene (4 mentions) Anti α tubulin, by Merck Group (4 mentions) Anti cyr61, by Cell Signaling Technology (2 mentions) Ma5 14154, by Thermo Fisher Scientific (2 mentions) Anti id2, by Cell Signaling Technology (2 mentions) Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Acetonitrile, by Avantor (1 mentions) Methanol, by Avantor (1 mentions) Amino acids, by Iris Biotech (1 mentions) Streptavidin horseradish peroxidase conjugate, by Thermo Fisher Scientific (1 mentions) Biotinylated human vegf a165, by R&D Systems (1 mentions) Formic acid, by Merck Group (1 mentions) Ammonium formate, by Merck Group (1 mentions) Anti atg 12 d88h11, by Cell Signaling Technology (1 mentions) Anti lc3bi 2 3868, by Cell Signaling Technology (1 mentions) Anti cleaved parp, by Cell Signaling Technology (1 mentions) Anti β actin, by Santa Cruz Biotechnology (1 mentions) Anti rabbit igg hrp, by Santa Cruz Biotechnology (1 mentions) Anti goat igg hrp, by Santa Cruz Biotechnology (1 mentions)

7 protocols using chemiluminescent

1

Peptide-receptor binding assay protocol

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Unless otherwise specified, reagents were obtained from commercial sources. LC-MS grade water was purchased from Huberlab (Aesch, Switzerland); methanol and acetonitrile (both HPLC gradient grade) were purchased from VWR (Darmstadt, Germany). Ammonium formate and formic acid were purchased from Sigma-Aldrich (Buchs, Switzerland). Other solvents and reagents were purchased from Merck (Darmstadt, Germany). Fmoc-Arg(Pbf) Wang resin was obtained from Activotec (Cambridge, UK). Amino acids and coupling reagents were purchased from Iris Biotech (Marktredwitz, Germany). Recombinant human receptors and biotinylated human VEGF-A165 were purchased from R&D Systems (Minneapolis, MN, USA). Chemiluminescent, streptavidin-horseradish peroxidase conjugate and DPBS were obtained from Thermo Scientific (Waltham, MA, USA).
Puszko A.K., Sosnowski P., Rignault-Bricard R., Hermine O., Hopfgartner G., Pułka-Ziach K., Lepelletier Y, & Misicka A. (2020). Urea-Peptide Hybrids as VEGF-A165/NRP-1 Complex Inhibitors with Improved Receptor Affinity and Biological Properties. International Journal of Molecular Sciences, 22(1), 72.
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2

Western Blot Analysis of Autophagy Markers

Cited 2 times
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Cells were lysed using 1% Triton in TBS containing protease and phosphatase inhibitors. Tumors were lysed using 1X RIPA buffer containing protease and phosphatase inhibitors. Equal amounts of total protein were separated by electrophoresis on a 4–12% Bis-Tris gel and transferred to a PVDF membrane (23 (link)). Blots were blocked in 5% BSA, and then probed with antibodies against anti- NURR1 (sc-376984, SCB), anti- ATG 7 (10088-2AP, Proteintech), anti-ATG 12(D88H11, Cell Signaling Technology), anti-LC3BI/II(3868, Cell Signaling Technology), anti-Cleaved PARP (9532S, Cell Signaling Technology) and anti-α-tubulin (Sigma). Chemiluminescent (32106, Thermo Fisher Scientific) signal was captured using a Syngene G-BOX iChemi XT imager (26 (link)).
Zarei M., Shrestha R., Johnson S., Yu Z., Karki K., Vaziri-Gohar A., Epps J., Du H., Suva L., Zarei M, & Safe S. (2021). Nuclear Receptor 4A2 (NR4A2/NURR1) Regulates Autophagy and Chemoresistance in Pancreatic Ductal Adenocarcinoma. Cancer Research Communications, 1(2), 65-78.
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3

Immunoblotting Antibody Detection Protocol

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Antibodies used for standard immunoblotting were: anti-p21 (Santa Cruz Biotechnology, sc-469; 1:250), anti- β-actin (Santa Cruz Biotechnology, sc-1616; 1:500), anti-rabbit IgG-HRP (Santa Cruz Biotechnology, sc-2004; 1:5,000), anti-goat IgG-HRP (Santa Cruz Biotechnology, sc-2020; 1:5,000). Signals were detected with “chemiluminescent” (Thermo Fisher Scientific). After the detection of p21, the same blot was stripped and reprobed with the anti-β-actin antibody as a loading control (see Supplementary Fig. 7b for uncropped scans of the blot).
Kogut I., McCarthy S.M., Pavlova M., Astling D.P., Chen X., Jakimenko A., Jones K.L., Getahun A., Cambier J.C., Pasmooij A.M., Jonkman M.F., Roop D.R, & Bilousova G. (2018). High-efficiency RNA-based reprogramming of human primary fibroblasts. Nature Communications, 9, 745.
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4

Western Blot Analysis of IDH1 Protein

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Cells were lysed using 1X ice-cold RIPA buffer supplemented with protease and phosphatase inhibitors. Protein concentration was quantified using the Pierce™ B.C.A. Protein Assay kit (Thermo Fischer Scientific). Equal amounts of whole protein extracts were loaded and separated by electrophoresis on a 4–12% Bis-Tris gel (Life Technologies) and transferred to a PVDF membrane (Thermo Fischer Scientific). Blots were blocked in 5% (wt/vol) non-fat milk and then probed with antibodies against anti-IDH1 (Invitrogen, OTI2H9) and anti-beta-actin (Invitrogen, 15739-BTIN). Chemiluminescent (32106, Thermo Fisher Scientific) signal was captured using a digital imager (Odyssey Infrared Imaging system).
Zarei M., Hajihassani O., Hue J.J., Graor H.J., Rothermel L.D, & Winter J.M. (2023). Targeting wild-type IDH1 enhances chemosensitivity in pancreatic cancer. bioRxiv.
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5

MITF, CYR61, and ID2 Protein Expression

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Cells were lysed using 1% Triton in TBS containing protease and phosphatase inhibitors. Tumors were lysed using 1X RIPA buffer containing protease and phosphatase inhibitors. Equal amounts of total protein were separated by electrophoresis on a 4–12% Bis-Tris gel and transferred to a PVDF membrane. Blots were blocked in 5% BSA, and then probed with antibodies against anti-MITF (MA5-14154, ThermoFisher Scientific), anti-CYR61 (14479, Cell Signaling Technology), anti-ID2 (3431, Cell Signaling Technology), and anti-α-tubulin (Sigma). Chemiluminescent (32106, Thermo Fisher Scientific) signal was captured using a Syngene G-BOX iChemi XT imager.
Zarei M., Giannikou K., Du H., Liu H.J., Duarte M., Johnson S., Nassar A.H., Widlund H.R., Henske E.P., Long H.W, & Kwiatkowski D.J. (2020). MITF is a driver oncogene and potential therapeutic target in kidney angiomyolipoma tumors through transcriptional regulation of CYR61. Oncogene, 40(1), 112-126.
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6

MITF, CYR61, and ID2 Protein Expression

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Cells were lysed using 1% Triton in TBS containing protease and phosphatase inhibitors. Tumors were lysed using 1X RIPA buffer containing protease and phosphatase inhibitors. Equal amounts of total protein were separated by electrophoresis on a 4–12% Bis-Tris gel and transferred to a PVDF membrane. Blots were blocked in 5% BSA, and then probed with antibodies against anti-MITF (MA5-14154, ThermoFisher Scientific), anti-CYR61 (14479, Cell Signaling Technology), anti-ID2 (3431, Cell Signaling Technology), and anti-α-tubulin (Sigma). Chemiluminescent (32106, Thermo Fisher Scientific) signal was captured using a Syngene G-BOX iChemi XT imager.
Zarei M., Giannikou K., Du H., Liu H.J., Duarte M., Johnson S., Nassar A.H., Widlund H.R., Henske E.P., Long H.W, & Kwiatkowski D.J. (2020). MITF is a driver oncogene and potential therapeutic target in kidney angiomyolipoma tumors through transcriptional regulation of CYR61. Oncogene, 40(1), 112-126.
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7

Immunoblotting Protein Detection Protocol

Cited 1 time
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Immunoblotting to detect protein detection was performed using SDS-PAGE as previously described (Lal et al., 2014 (link)). Membranes for immunoblotting were probed with antibodies against anti-phospho-CTD-RNAPII-S2 (04-1571; Millipore), anti-phospho-CTD-RNAPII-S5 (04-1572; Millipore), anti-CDK7 (2916; Cell Signaling Technology), anti-CDK13 (A301-458A; Bethyl), anti-cleaved poly(ADP ribose) polymerase (Asp214; 9541; Cell Signaling Technology), anti-cleaved caspase 3 (Asp175; 9664; Cell Signaling Technology), NRF2 (12721; Cell Signaling Technology), and anti-α-tubulin (Sigma-Aldrich). Chemiluminescent (32106; Thermo Fisher Scientific) signal was captured using a Syngene G-BOX iChemi XT imager.
Zarei M., Du H., Nassar A.H., Yan R.E., Giannikou K., Johnson S.H., Lam H.C., Henske E.P., Wang Y., Zhang T., Asara J, & Kwiatkowski D.J. (2019). Tumors with TSC mutations are sensitive to CDK7 inhibition through NRF2 and glutathione depletion. The Journal of Experimental Medicine, 216(11), 2635-2652.
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