Proteins were separated on 4–20% gradient SERVA gels and transferred onto a nitrocellulose membrane using a Trans‐Blot® Turbo™ Transfer System (Bio‐Rad). SIGMAFAST™ BCIP®/NBT or SignalFire™ Elite ECL Reagent were used to visualise proteins on C‐DiGit Chemiluminescent Blot Scanner (LI‐COR Biosciences). The following antibodies were used: custom polyclonal antibody raised against Lsr2 in rabbit (Gemini Biosciences); monoclonal murine anti‐polyhistidine antibody (Sigma‐Aldrich); phospho‐threonine antibody (Cell Signaling Technology); monoclonal anti‐MtbGroEL2 (Rv0440), clone IT‐70 (BEIResources); mouse anti‐rabbit IgG antibody:alkaline phosphatase (Sigma‐Aldrich; anti‐mouse IgG (whole molecule:alkaline phosphatase antibody produced in rabbit (Sigma‐Aldrich), and anti‐rabbit IgG, HRP‐linked antibody (Cell Signaling Technology).
C digit chemiluminescent blot scanner
Manufactured by LI COR
Sourced in United States, United Kingdom
The C-DiGit Chemiluminescent Blot Scanner is a laboratory equipment designed to capture and analyze chemiluminescent signals from blots. It provides high-resolution digital imaging and quantitative analysis of chemiluminescent western blots, northern blots, and other similar applications.
Automatically generated - may contain errors
Lab products found in correlation
Trans blot turbo transfer system, by Bio-Rad (4 mentions)
Sigmafast bcip nbt, by LI COR (3 mentions)
Signalfire elite ecl reagent, by LI COR (3 mentions)
Monoclonal murine anti polyhistidine antibody, by Merck Group (2 mentions)
Phospho threonine antibody, by Cell Signaling Technology (2 mentions)
Mouse anti rabbit igg antibody alkaline phosphatase, by Merck Group (2 mentions)
Anti mouse igg whole molecule alkaline phosphatase antibody, by Merck Group (2 mentions)
Anti rabbit igg hrp linked antibody, by Cell Signaling Technology (2 mentions)
Bca protein assay, by Thermo Fisher Scientific (2 mentions)
Image studio software, by LI COR (2 mentions)
Horseradish peroxidase hrp conjugated secondary antibody, by Jackson ImmunoResearch (1 mentions)
Supersignal west femto ecl, by Thermo Fisher Scientific (1 mentions)
Gapdh, by Merck Group (1 mentions)
Pvdf membrane, by Thermo Fisher Scientific (1 mentions)
Halt protease inhibitor cocktail, by Thermo Fisher Scientific (1 mentions)
Ab1191, by Abcam (1 mentions)
Ab15823, by Abcam (1 mentions)
P0448, by Agilent Technologies (1 mentions)
Ab1791, by Abcam (1 mentions)
Amersham protran premium 0.45 μm nitrocellulose membrane, by GE Healthcare (1 mentions)
8 protocols using c digit chemiluminescent blot scanner
1
Immunoblotting of Mycobacterial Proteins
2
Brain Protein Extraction and Analysis
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Proteins were extracted from sagittal half-brain homogenates using a buffer containing 150 mM NaCl, 50 mM Tris-HCl, 1% Triton X-100, 5 mM EDTA, 0.1% SDS, and 1% sodium deoxycolate freshly supplemented with Halt Protease Inhibitor Cocktail (Thermo Scientific, Rockford, IL, USA), and they were measured by BCA Protein Assay (Thermo Scientific). Proteins were separated by gradient SDS-PAGE (4%–20% TGX, Bio-Rad) and blotted onto polyvinylidene fluoride (PVDF) membrane (Invitrogen). Western blots were probed with antibodies to PLP/DM20 (1:2,000, AA3 Hybridoma) and GAPDH (1:50,000, Sigma-Aldrich), followed by horseradish peroxidase (HRP)-conjugated secondary antibody (1:5,000, Jackson ImmunoResearch), and detected using SuperSignal West Femto ECL (Thermo Scientific). Bands were visualized using C-DiGit Chemiluminescent Blot Scanner (LI-COR Biosciences, Lincoln, NE, USA) and quantified using iS Image Studio Version 4.0 (LI-COR Biosciences).
3
Histone Acetylation Analysis in Drosophila
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For sample preparation Drosophila embryos were homogenized with a plastic pestle in sonication buffer (50 mM Tris-HCl pH7.9, 2 mM EDTA, 50 mM NaCl, 0.5 mM DTT, 10 mM Na-butyrate and 1x Protease inhibitor cocktail set I (Calbiochem)), then sonicated on “high” energy setting for four cycles of 30 sec ON/30 sec OFF in a Bioruptor sonicator (Diagenode). Homogenized samples were boiled for 10 minutes in 2x Laemmli sample buffer containing 5% β-mercaptoethanol, then centrifuged for 10 minutes at 13000 RPM. Sample supernatants containing 30 μg protein were separated by 10% Tris-Tricine-SDS PAGE and electrotransferred to Amersham Protran Premium 0.45 μm nitrocellulose membrane (GE Healthcare Life Sciences). Membranes were blocked in 5% nonfat milk and incubated with the following primary and secondary antibodies in the indicated dilutions: anti-acetyl-H4K5 (ab61236, Abcam, 1:500), anti-acetyl-H4K8 (ab15823, Abcam, 1:1000), anti-acetyl-H4K12 (ab61238, Abcam, 1:1000), anti-acetyl-H3K18 (ab1191, Abcam, 1:500), anti-acetyl-H3K23 (ab47813, Abcam, 1:1000) anti-H3 (ab1791, Abcam, 1:4000), goat-anti-rabbit IgG-HRP (P0448, Dako, 1:4000). Immunoblots were developed with Immobilon Western Chemiluminescent HRP substrate (Millipore) and recorded with a C-DiGit chemiluminescent blot scanner (Li-Cor Biosciences). Band intensities were quantitated with Image Studio software (Li-Cor Biosciences).
4
Western Blot Analysis of Diagnostic Proteins
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The proteins were separated on 4% to 20% gradient SERVA gels and transferred onto a nitrocellulose membrane using a Trans-Blot Turbo transfer system (Bio-Rad) according to the manufacturer’s instructions. SignalFire Elite ECL reagent (Cell Signaling, UK) was used to visualize proteins on a C-DiGit chemiluminescent blot scanner (LI-COR Biosciences), according to the manufacturer’s instructions. All the secondary antibodies were from Cell Signaling, UK. Diagnostic proteins were used for all the cellular fractions: GlnA (membrane protein), GarA (secreted and cytoplasmic protein), RpfB (membrane and cell wall protein). and FtsZ (cytoplasmic protein).
5
Western Blot Analysis of Lsr2 Protein
6
Protein Separation and Visualization
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Proteins were separated on 4%–20% gradient SERVA gels and transferred onto a nitrocellulose membrane using a Trans-Blot® Turbo Transfer System (Bio-Rad). SIGMAFAST BCIP®/NBT or SignalFire Elite ECL Reagent were used to visualize proteins on C-DiGit Chemiluminescent Blot Scanner (LI-COR Biosciences), according to the manufacturer’s instructions.
7
Western Blot Analysis of Exosome Proteins
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Exosome protein lysates were resolved on Tris-Glycine SDS-PAGE and transferred to polyvinylidene difluoride (Immobilon-P; Millipore) membranes. Membranes were incubated in primary antibody (Abcam anti-TSG101 clone 4A10, diluted 1:100 per manufacturer instructions) overnight at 4°C, washed three times with 0.1% TBST, incubated with secondary antibody (HRP-conjugated goat anti-mouse, Thermo Scientific, Pierce, 1:10,000) for 1 h at room temperature, washed three times and detected with enhanced chemiluminescence (PerkinElmer LLC or Life Technologies) on LI-COR C-DiGit Chemiluminescent Blot Scanner using Image Studio Digits software Ver 3.1
8
Western Blot Analysis of YAP Signaling
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Cells were harvested in chilled RIPA Cell Lysis Buffer with EDTA (GenDEPOT, Barker, TX, R4100‐010) with 1% protease and phosphatase inhibitor cocktails (Halt, ThermoFisher, Waltham, MA). After protein determination and normalization between conditions, SDS‐PAGE was performed using Tris‐HCl poured gels. Electrophoretic transfer from gel to nitrocellulose blot was performed with the eBlot L1 Transfer System (GenScript, Piscataway, NJ). Western blotting was performed using antibodies against YAP (Santa Cruz, Dallas, TX, clone H‐9, Cat. No. sc‐271134), phospho‐YAP (Ser127; Cell Signaling Technology, Danvers, MA, Cat. No. 4911), β‐actin (Santa Cruz, Dallas, TX, clone C4, Cat. No. sc‐47778), GAPDH (Cell Signaling Technology, Danvers, MA, 14C10, Rabbit mAb #2118), HA tag (Santa Cruz, Dallas, TX, clone F‐7, Cat. No. sc‐7392), and DYKDDDDK Tag (FLAG; Cell Signaling Technology, Danvers, MA, Cat. No. 2368S). After secondary HRP, Western Sure Chemiluminescent substrate (LI‐COR, Lincoln, NE) was applied. The LI‐COR C‐DiGit chemiluminescent blot scanner was used to scan blots and the Image Studio software (LI‐COR, Lincoln, NE) to quantify band intensities between conditions.
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