Cx43MP (250 µg/mL) was prepared in HCl (1 M) and NaOH (1 M), respectively. Aliquots were kept at 37 °C for 24 h (Thermo Scientific Heraeus® microbiological incubator, USA) and 80 °C (AccuBlock™ Digital Dry Baths, Labnet international, USA) for 12 h.
Accublock digital dry bath
Manufactured by Labnet
Sourced in United States
The AccuBlock™ Digital Dry Bath is a laboratory instrument designed for precise temperature control. It provides a consistent and reliable heating platform for a variety of sample types and applications.
Automatically generated - may contain errors
Lab products found in correlation
Geneatlas imaging station, by Thermo Fisher Scientific (3 mentions)
Geneatlas fluidics station, by Thermo Fisher Scientific (2 mentions)
Gene chip human genome u219 microarrays, by Thermo Fisher Scientific (2 mentions)
Heraeus, by Thermo Fisher Scientific (1 mentions)
Genechip 3 ivt plus reagent kit, by Thermo Fisher Scientific (1 mentions)
8 protocols using accublock digital dry bath
1
Thermal Stability Evaluation of Cx43MP
2
Rapid E. coli DNA Extraction via Boiling
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Escherichia coli was resuscitated from glycerol stock and plated on EMB plates. Single colonies were picked from the individual EMB plates and inoculated into the nutrient broth and incubated for 48 hours at 37°C. DNA was extracted by the boiling method [10 ]. Briefly, 1 ml of broth solution containing E. coli was transferred to an Eppendorf tube and centrifuged (Thermo Fisher Scientific, Germany) for 15 mins at a speed of 13000 rpm. The supernatant was discarded, and the pellet was retained. Again another 1 ml of the broth was added and centrifuged at the same speed and duration; this was done five times to obtain a sizeable pellet. To wash the pellet; 200 μL of distilled water was added to the pellet and discarded. Again a volume of 200 μL of distilled water was added to the washed pellet, vortexed and centrifuged at a speed of 13000 rpm for 5 mins and the supernatant was discarded. The pellet was then placed on AccuBlock™ Digital Dry Baths (Labnet International, USA) at 100°C for 15 mins to lyse the cell. Cell debris was removed by centrifugation at 13000 rpm for 10 mins while the supernatant was stored as the DNA template.
3
Affymetrix GeneChip U219 Microarray Protocol
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The samples were loaded onto and hybridized to Affymetrix GeneChip Human Genome U219 microarrays, together with control cRNA and oligo B2. Hybridization was conducted at 45°C for 16 hours. using an AccuBlock™ Digital Dry Bath (Labnet International, Inc. NY, USA) hybridization oven Next, the microarrays were washed and stained according to manufacturer's protocol using an Affymetrix GeneAtlas™ Fluidics Station (Affymetrix, Santa Clara, CA, USA), and the chips were scanned using an Affymetrix GeneAtlas™ Imaging Station (Affymetrix, Santa Clara, CA, USA). The scans of the microarrays were saved as *.CEL files on hard disks.
4
Affymetrix GeneChip Array Protocol
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Biotin-labelled and fragmented target cRNA samples were loaded into Affymetrix GeneChip (Human Genome U219) Array Strips together with controls cRNAs and oligo B2. The hybridisation procedure was conducted at 45°C for 16 h in an AccuBlock Digital Dry Bath (Labnet international, Inc.) hybridisation oven. The washing and staining procedure was performed using an Affymetrix GeneAtlas Fuidics Station according to the instructions in the technical manual. An Affymetrix GeneAtlas Imaging Station was used for scanning the arrays.
5
Microarray Gene Expression Profiling Protocol
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To conduct microarray experiments, 100 ng of total RNA of each condition was used as input. Samples were reversely transcribed into cDNA and then in vitro transcribed according to the manufacturer’s protocol using GeneChip™ 3′ IVT PLUS Reagent Kit (Applied Biosystems, Thermo Fisher Scientific Inc.). Next, samples were fragmented and hybridized on Affymetrix GeneChip human genome U219 microarrays, together with control cRNA and oligo B2. Hybridization was conducted at 45 °C for 16 h, using an AccuBlock™ Digital dry bath (Labnet International, Inc., New York, NY, USA) hybridization oven. Further, the microarrays were washed and stained according to the manufacturer’s protocol using an Affymetrix GeneAtlas™ Fluidics Station (Affymetrix, Santa Clara, CA, USA). In the final step, all microarrays were scanned using an Affymetrix GeneAtlas™ imaging station (Affymetrix). The scans of the microarrays were saved as *.CEL files on local storage. Downstream analysis was conducted in R/RStudio IDE (R – version 4.0.2, RStudio – version 1.3.1056), Bioconductor (version 1.3.10) as well as Transcriptome Analysis Software 4.0.1.36. Microarray results were stored in the GEO database under ID GSE140996.
6
Activating Latent TGF-β via Heat
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It became necessary to verify the integrity of the latent TGF-β, and to test whether or not it was possible to release the active TGF-β from the small latent complex using only heat. To this end, latent TGF-β was split into two groups: an experimental group that would be heated on an AccuBlock Digital Dry Bath (Labnet International, Inc., USA) at 80 °C for 10 min, and a control group kept at room temperature. The resulting release of active TGF-β was then quantified via ELISA.
7
Fluorescent Peptide Biosensor Assay
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The emission spectrometry measurements of the 7-amino-4-methylcoumarin (AMC) peptide substrate were obtained using Shimazdu Scientific RF-5301 spectrofluorophotometer (Portland, OR, USA). The substrate solutions 0–0.5 μmol/mL were prepared using the PBS solution and 100 μL were added to the centrifuge tube. A 2 mg sample of biosensor tNFC-Pep was placed into a centrifuge tube with 100 μL of PBS. A volume of 100 μL of the elastase enzyme 1 U/mL was added to the wells containing the substrate and biosensors for a total volume of 200 μL. The centrifuge tube was placed onto an Accu block digital dry bath (Labnet International, Edison, NJ, USA) for 30 min at 37 °C. These samples were diluted (15×) with PBS, 3 mL of each sample were placed in a cuvette, and fluorescence measurements were scanned from 405–625 nm with an excitation at 390 nm.
8
Affymetrix GeneChip® Hybridization Workflow
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Biotin-labeled and fragmented target cRNA samples were loaded into Affymetrix GeneChip® (Human Genome U219) Array Strip together with control cRNAs and oligo B2. Hybridization procedure was conducted at 45 °C, for 16 h in AccuBlock™ Digital Dry Bath (Labnet international, Inc.) hybridization oven. Washing and staining procedure was performed in Affymetrix GeneAtlas™ Fuidics Station according to the instructions in the technical manual. Affymetrix GeneAtlas™ Imaging Station was used for scanning the arrays.
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