Log In Sign up
The largest database of trusted experimental protocols

Pico chemiluminescent substrate

Manufactured by Thermo Fisher Scientific
Visit Supplier
Sourced in United States

The Pico Chemiluminescent Substrate is a laboratory reagent used to detect and quantify proteins in Western blot analysis. It generates a luminescent signal proportional to the amount of target protein present, enabling sensitive and quantitative protein detection.

Automatically generated - may contain errors

Lab products found in correlation

Pvdf membrane, by Merck Group (5 mentions) Bca protein assay kit, by Thermo Fisher Scientific (4 mentions) Protease inhibitor cocktail, by Serva Electrophoresis (3 mentions) Triton x 100, by Serva Electrophoresis (3 mentions) Phenylmethylsulfonyl fluoride, by Merck Group (3 mentions) Gradient sds page gel, by Bio-Rad (2 mentions) Anti rab27, by Santa Cruz Biotechnology (2 mentions) Anti cd63, by BD (2 mentions) Anti bip, by BD (2 mentions) Anti cd81, by Santa Cruz Biotechnology (2 mentions) Anti tsg101, by Abcam (2 mentions) Sigma ge28 9068 38, by Merck Group (2 mentions) Anti cd9, by Abcam (2 mentions) Recombinant egfp, by Abcam (2 mentions) Protease inhibitor, by Roche (2 mentions) Supersignal west femto, by Thermo Fisher Scientific (2 mentions) Hrp conjugated secondary antibody, by Santa Cruz Biotechnology (1 mentions) Ab200997, by Abcam (1 mentions) Ab205718, by Abcam (1 mentions) Ab82411, by Abcam (1 mentions)

18 protocols using pico chemiluminescent substrate

1

Western Blot Analysis of IRF3

Check if the same lab product or an alternative is used in the 5 most similar protocols
Western blots were performed by lysing cells using ice-cooled lysis buffer (20 mM Tris/HCl, pH 7.5, 150 mM NaCl, 1 mM phenylmethylsulfonyl fluoride (Sigma-Aldrich), protease inhibitor cocktail, 1 Triton X-100 (Serva, Mannheim, Germany), and 10 % glycerol). Lysates were analyzed using SDS-PAGE gels (Novex NuPAGE 10 % Bis-Tris Protein Gel, Life Technologies). Proteins were transferred to PVDF membrane (Millipore, Billerica, MA, USA) using wet blotting. IRF3 was detected using the primary antibody clone D83B9 (Cell Signaling Technology, Danvers, MA, USA). Secondary anti-rabbit antibodies were HRP-conjugated (Dianova, Hamburg, Germany). Detection was performed using the Pico Chemiluminescent Substrate from Thermo Scientific (Asheville, NC, USA) and a CCD camera.
Liesche C., Venkatraman L., Aschenbrenner S., Grosse S., Grimm D., Eils R, & Beaudouin J. (2016). Death receptor-based enrichment of Cas9-expressing cells. BMC Biotechnology, 16, 17.
+ Open protocol
+ Expand
2

Extracellular Vesicle Protein Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
EV-containing fractions were lysed in 1X RIPA lysis buffer. Protein concentrations were determined by BCA protein assay kit (Thermo Fisher). Equivalent total protein amounts from BH and EVs were separated on 4 − 15% stain-free pre-cast SDS-PAGE gradient gels (Bio-Rad) under non-reducing conditions and transferred onto PVDF membranes (Sigma Aldrich). After 1 h blocking in 5% non-fat milk solution (Bio-Rad 170–6404) at room temperature, membranes were incubated with anti-CD63 (1:1000 dilution), anti-Bip (1:1000 dilution) (BD Biosciences 556019 and 610978, respectively), anti-CD81 (1:1000 dilution), anti-Rab27 (1:1000 dilution) (Santa Cruz Biotechnology sc23962, sc74586), anti-TSG101 (1:500 dilution), anti-CD9 (1:500 dilution), anti-Syntenin (1:500 dilution), anti-Calnexin (1:2000 dilution), or anti-GM130 (1:1000 dilution) (the last five antibodies were Abcam ab125011, ab92726, ab133267, ab22595, and ab76154) overnight at 4°C. The membrane was washed 3 times for 8 min in PBST while shaking, then incubated with HRP-conjugated secondary antibody (1:10000 dilution) (Santa Cruz Biotechnology sc-2357, sc-516102) at room temperature for 1 h. After washing again in PBST, the enzyme-linked antibody was detected by incubation with Pico chemiluminescent substrate (Thermo Fisher 34580) and recording on film (Millipore Sigma GE28-9068-38).
Huang Y., Cheng L., Turchinovich A., Mahairaki V., Troncoso J.C., Pletniková O., Haughey N.J., Vella L.J., Hill A.F., Zheng L, & Witwer K.W. (2020). Influence of species and processing parameters on recovery and content of brain tissue-derived extracellular vesicles. Journal of Extracellular Vesicles, 9(1), 1785746.
+ Open protocol
+ Expand
3

Quantitative Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Total protein concentration in the plasma samples was determined using Pierce BCA Protein Assay (Thermo Fisher, Waltham, MA). General protein separation was carried out by electrophoresis on 4–20% gradient SDS-PAGE gels at 110 V for 1.25 h and transferred to nitrocellulose membranes (Bio-Rad Laboratories, Inc., Hercules, CA). Transfer membranes were then incubated in blocking buffer (1% BSA in TBST) at 4 °C under gentle agitation overnight. Blocked membranes were probed with primary recombinant monoclonal antibodies for trypsin (ab200997, 1:1000, Abcam, Cambridge, MA), pancreatic lipase (ab124915,1:30000, Abcam), and Transferrin (ab82411, 1:10,000, Abcam) before incubated in the corresponding secondary antibody (ab205718, 1/5000, Abcam). Membranes were incubated with Pico Chemiluminescent Substrate (Thermo Fisher Scientific) and then imaged via photo-sensitive autoradiography film (Genesee Scientific, San Diego, CA). Western blot images were digitally analyzed (gel analysis tool, Image J; NIH). The band density values for each of the western blots for trypsin and lipase were normalized to their respective transferrin values.
Courelli V., Ahmad A., Ghassemian M., Pruitt C., Mills P.J, & Schmid-Schönbein G.W. (2021). Digestive Enzyme Activity and Protein Degradation in Plasma of Heart Failure Patients. Cellular and Molecular Bioengineering, 14(6), 583-596.
+ Open protocol
+ Expand
4

Quantification of Endogenous Proteins by Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Western blot samples were lysed with lysis buffer [20 mM tris-HCl (pH 7.5), 150 mM NaCl, 1 mM phenylmethylsulfonyl fluoride (Sigma-Aldrich), protease inhibitor cocktail, 1% Triton X-100 (Serva), and 10% glycerol]. Lysates were analyzed with SDS–polyacrylamide gel electrophoresis gels (Invitrogen). The different time points were loaded in a random manner to avoid quantification bias (figs. S7 and S8). Proteins were transferred onto a polyvinylidene difluoride (PVDF) membrane (Millipore) by wet blotting. Detection was performed with the Pico Chemiluminescent Substrate from Thermo Scientific and a charge-coupled device camera (Intas).
In probe-expressing CD95-HeLa and wild-type HeLa cell lines that were used for time series experiments, we quantified endogenous FADD, p55, and BID concentrations using calibrated GFP fusion proteins (figs. S9 to S11). First, GFP fusion protein lysates were calibrated to recombinant eGFP (BioVision). We combined equal amounts of lysates of nontransfected cells with lysates of the fusion protein–expressing cells. This mixed lysate was combined with different amounts (0.1 and 30 ng) of GFP and then loaded onto Western blot gels. Different amounts of these calibrated fusion protein lysates were loaded onto Western blot gels along with lysates from the cleavage probe–expressing cell lines that were used for time series experiments.
Kallenberger S.M., Beaudouin J., Claus J., Fischer C., Sorger P.K., Legewie S, & Eils R. (2014). Intra- and Interdimeric Caspase-8 Self-Cleavage Controls Strength and Timing of CD95-Induced Apoptosis. Science signaling, 7(316), ra23.
+ Open protocol
+ Expand
5

Western Blot Analysis of Connexin 43 and pSMAD1/5/9

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were lysed in RIPA buffer (20mM Tris-HCl, 0.1% SDS, 1% Triton X-100, 1% sodium deoxycholate) containing protease inhibitor (Roche). The resulting lysates were run on 10% Mini-PROTEAN® TGX™ Gel (Bio-Rad) and transferred to Amersham Hybond-P membrane (GE Healthcare). The following antibodies were used (all are from Cell Signaling Technology): rabbit anti-connexin 43 (1:500, Cat. no. 3512, RRID: AB_2294590), rabbit anti-pSMAD1/5/9 (1:1000, Cat. no. 13820, RRID: AB_2493181), and rabbit anti-GAPDH (1:2000, Cat. no. 2118, RRID: AB_1031003). SuperSignal West Femto Chemiluminescent substrate was used to detect anti-connexin 43 and anti-pSMAD1/5/9, and Pico Chemiluminescent substrate (Thermo Scientific) was used to detect anti-GAPDH.
Shi C., Zhang H., Louie K., Mishina Y, & Sun H. (2016). BMP Signaling Mediated by BMPR1A in Osteoclasts Negatively Regulates Osteoblast Mineralization Through Suppression of Cx43. Journal of cellular biochemistry, 118(3), 605-614.
+ Open protocol
+ Expand
6

Western Blot Protein Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Preparation of protein lysates, SDS-PAGE, transfer to PVDF membranes (Hybond-P; Amersham), and antibody incubations were performed using standard procedures. Blots were developed using SuperSignal West Femto or Pico Chemiluminescent Substrate (both from Thermo Scientific) and scanned using a ChemiDoc Touch Imaging System (Bio Rad). Densitometric analysis was performed with Image-J software (National Institutes of Health, Maryland, USA).
Nguyen C.H., Glüxam T., Schlerka A., Bauer K., Grandits A.M., Hackl H., Dovey O., Zöchbauer-Müller S., Cooper J.L., Vassiliou G.S., Stoiber D., Wieser R, & Heller G. (2019). SOCS2 is part of a highly prognostic 4-gene signature in AML and promotes disease aggressiveness. Scientific Reports, 9, 9139.
+ Open protocol
+ Expand
7

Quantitative Immunoblot Analysis of EpoR-GFP

Check if the same lab product or an alternative is used in the 5 most similar protocols
Immunoblot samples were lysed with lysis buffer (20 mM Tris/HCl, pH 7.5, 150 mM NaCl, 1 mM phenylmethylsulfonyl fluoride (Sigma-Aldrich), protease inhibitor cocktail, 1% Triton X-100 (Serva, Mannheim, Germany), and 10% glycerol). Cell lysates were analyzed using SDS PAGE gels (Invitrogen). Proteins were transferred to PVDF membrane (Merck Millipore) using wet blotting. Detection was performed using the Pico Chemiluminescent Substrate from Thermo Scientific and a CCD camera (Intas, Göttingen, Germany). EpoR-GFP concentrations in EpoR-GFP-expressing H838 cells were quantified utilizing recombinant eGFP (BioVision, Mountain View, CA, USA). Cell lysates were combined with different amounts of GFP ranging from 0.2 to 10 ng and then loaded onto gels (S3 Fig). To detect EpoR-GFP and GFP in immunoblots, we used an antibody recognizing GFP (clones 7.1 and 13.1) from Roche (Basel, Switzerland). Horseradish peroxidase-conjugated secondary antibodies (Southern Biotech, Birmingham, AL, USA) were used for detection.
Kallenberger S.M., Unger A.L., Legewie S., Lymperopoulos K., Klingmüller U., Eils R, & Herten D.P. (2017). Correlated receptor transport processes buffer single-cell heterogeneity. PLoS Computational Biology, 13(9), e1005779.
+ Open protocol
+ Expand
8

Western Blotting Protocol for Protein Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Whole-cell lysates were obtained by resuspending cell pellets in RIPA buffer (50 mM Tris pH 7.4, 150 mM NaCl, 1% Triton X-100) with freshly added protease inhibitor (Roche). The concentration of protein was measured by a BCA protein assay kit (Pierce, Rockford, IL, USA) according to the manufacturer’s instructions. 50 µg of protein lysate was fractionated by SDS-PAGE and then transferred onto a PVDF membrane. The membrane was blocked with 5% milk at room temperature for 1 h then probed with primary antibodies overnight at 4°C. The membrane was washed with TBST and then incubated with corresponding secondary antibody for 1 h at room temperature. The targeted proteins were visualized with super signal West Pico Chemiluminescent Substrate (Thermo Scientific, USA). The anti-β-actin antibody (Sigma, Cat #A2228) was used as an internal control. Densitometry was performed using Image J (National Institutes of Health, USA). Protein levels were normalized to β-actin.
Wu G., Xu G., Chen D.W., Gao W.X., Xiong J.Q., Shen H.Y, & Gao Y.Q. (2018). Hypoxia Exacerbates Inflammatory Acute Lung Injury via the Toll-Like Receptor 4 Signaling Pathway. Frontiers in Immunology, 9, 1667.
+ Open protocol
+ Expand
9

Protein Detection by Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cell lysate with equal amount of protein was resolved by SDS-PAGE, and then transferred to NC membrane. After being blocked by 5% nonfat milk, the membrane was incubated with primary and secondary antibodies sequentially. Signals were developed by Pico Chemiluminescent Substrate (Thermo Fisher Scientific, USA) on films.
Feng H., Wang J., Chen W., Shan B., Guo Y., Xu J., Wang L., Guo P, & Zhang Y. (2016). Hypoxia-induced autophagy as an additional mechanism in human osteosarcoma radioresistance. Journal of Bone Oncology, 5(2), 67-73.
+ Open protocol
+ Expand
10

Western Blot Analysis of Epigenetic Regulators

Check if the same lab product or an alternative is used in the 5 most similar protocols
For western blot, proteins were separated on a denaturing SDS-PAGE gel and transferred to PVDF membrane. Membranes were blocked in PBST +5% nonfat dry milk at room temp for >10 min or at 4°C overnight. Primary antibodies used for western blot were: FLAG M2 monoclonal antibody (Sigma Aldrich F1804), TET2 monoclonal antibody (Millipore MABE462), TET3 polyclonal antibody (Millipore ABE383), OGT polyclonal antibody (Santa Cruz sc32921), OGT monoclonal antibody (Cell Signaling D1D8Q), His6 monoclonal antibody (Thermo MA1-21315), JL8 GFP monoclonal antibody (Clontech), and O-GlcNAc RL2 monoclonal antibody (Abcam ab2739). Secondary antibodies used were goat anti-mouse HRP and goat anti-rabbit HRP from BioRad. Blots were incubated with Pico Chemiluminescent Substrate (ThermoFisher) and exposed to film in a dark room.
Hrit J., Goodrich L., Li C., Wang B.A., Nie J., Cui X., Martin E.A., Simental E., Fernandez J., Liu M.Y., Nery J.R., Castanon R., Kohli R.M., Tretyakova N., He C., Ecker J.R., Goll M, & Panning B. (2018). OGT binds a conserved C-terminal domain of TET1 to regulate TET1 activity and function in development. eLife, 7, e34870.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!