To investigate the effects of the treatments on protein expression, posttreatment psoriatic rat skin (3 rats/group) was collected from the 10% sericin (the most effective dose of sericin), nontreatment, betamethasone, and calcitriol treatment groups. The samples were individually stored at −80 °C, and then, the samples were incubated in liquid nitrogen and homogenized in a mortar. The grind samples were solubilized in 300 μl of lysis buffer (1% SDS, 1% Triton-X, 0.5% NaCl) and ultrasonicated for 2 min with a 5-sec pulse on/off on ice. The samples were centrifuged at 10,000 g for 10 min at 4 °C, and the soluble proteins were collected. Total proteins were measured by a Bradford protein assay (Bio-Rad®, USA) using a spectrophotometer (Thermo Scientific, USA). Thirty micrograms of each crude protein sample was subjected to sodium dodecyl sulfate–polyacrylamide gel electrophoresis (SDS-PAGE) (4% stacking and 12% separating gels). The gel electrophoresis unit (Bio-Rad®, USA) was run with a constant voltage at 120 V for 90 min.
Gel electrophoresis unit
Manufactured by Bio-Rad
Sourced in United States
A gel electrophoresis unit is a laboratory instrument used to separate and analyze biomolecules, such as DNA, RNA, or proteins, based on their size and charge. The unit typically includes a gel-casting tray, an electrophoresis chamber, and a power supply. Samples are loaded into the gel, and an electrical current is applied, causing the biomolecules to migrate through the gel at different rates based on their characteristics. This process allows for the identification and quantification of specific biomolecules within a sample.
Automatically generated - may contain errors
5 protocols using gel electrophoresis unit
1
Sericin Impacts Psoriatic Skin Protein
2
Genotyping of Recombinant Inbred Lines
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Total genomic DNA from the RILs was extracted from young leaves using modified cetyl trimethyl ammonium bromide (CTAB) method [18 (link)]. The RILs were genotyped with the AhTE markers, which were polymorphic between TMV 2 and TMV 2-NLM (S1 Table ). The PCR was carried out in a reaction volume of 10 μl with 50 ng of template DNA, 5 pmol of each primer, 10X of Taq polymerase buffer [500 mM KCl, 100 mM Tris-HCI (pH 8.5], 2.0 mM of MgCl2, 0.25 mM of dNTPs and 0.15 U of Taq polymerase. PCR was performed in 96-wellplates using Veriti 96-Well Thermal Cycler (Applied Biosystem) with the temperature profile of 95°C for 5 min and 35 cycles of 95°C for 1 min, 53°C for 1 min and 72°C for 1.30 min, and 72°C for 8 min for final extension. The PCR products were analyzed by loading them on 1.8% agarose gel and electrophoresing in 1X TAE at 80 V for 2 h using Bio-Rad gel electrophoresis unit. The amplicons were visualized using ethidium bromide staining method. Specific PCR product was identified for each marker [9 (link), 12 (link)] and the alleles differing for 205 bp (equal to the size of AhMITE1) were scored. RILs were scored as A [homozygote as the first parent (TMV 2)], B [homozygote as the second parent (TMV 2-NLM)], H (heterozygote), C [not genotype a (b-allele is dominant)] and D [not genotype b (a-allele is dominant)] as per JoinMap 4 format [19 ],
3
Extending Coconut Water Shelf-life
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
In order to increase the shelf-life of coconut water, we intended to interact DNA with Ag nanoparticles, latter being regarded as a powerful antibacterial agent. At first, DNA was extracted from the coconut water, followed by isolation and then, subjected to run in the gel electrophoresis unit (BioRad, USA). The DNA isolation was carried out by following a standard protocol with certain modifications(Angeles et al., 2005 ). It is worth mentioning here that, since the mature endosperm has a high content of lipids and galactomannan contaminants, young leaves were chosen for DNA isolation by using a higher salt concentration (2 M) in the extraction buffer and polyvinyl poly pyrrolidone. The extracted DNA is then allowed to interact with different concentrations of green synthesized silver nano-particles (AgNPs). The concentration of AgNPs was varied in the range of 200–1000 ng/ml. Using Gel Electrophoresis, DNA-Ag NPs conjugates, native coconut DNA and control AgNPs were run simultaneously in different lanes for nearly 30 min at a potential difference of 120 mV.
After 30 min. of gel-run, it was placed in the non-UV based Gel-Doc instrument (BioRad, USA) for taking the image of the bands for analyses.
After 30 min. of gel-run, it was placed in the non-UV based Gel-Doc instrument (BioRad, USA) for taking the image of the bands for analyses.
4
Mulberry Leaves Impact on Plasma Proteome
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
To investigate the effect of mulberry leaves on protein expression in the plasma of people with impaired glucose metabolism, such as obese persons with prediabetes and patients with early-stage T2DM, the plasma specimens were used to quantify the concentration of total protein using a Bradford protein assay (Bio-Rad®, Hercules, CA, USA). For the SDS polyacrylamide gel electrophoresis (SDS-PAGE), a 4% stacking and 12% separating gel was loaded with 30 µg of each crude protein from the plasma samples and run with a constant voltage at 120 V for 80 min in the gel electrophoresis unit (Bio-Rad®, USA), until the bromophenol blue dye reached the bottom of the gel.
5
Transposable Element Genotyping in Peanut
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The genomic DNA from 60 genotypes was isolated from healthy young leaves using modified CTAB method (53 (link)). A total of 100 Arachis hypogaea transposable element (AhTE) markers were used to screen these genotypes (Supplementary Table 1 ). The PCR was carried out in a reaction volume of 10 μl with 50 ng of template DNA, 5 pmol of each primer, 10X of Taq polymerase buffer [500 mM KCl, 100 mM Tris-HCI (pH 8.5)], 0.25 mM of dNTPs and 0.1 U of Taq polymerase. The PCR reaction was carried out in 96 well plates using Mastercycler-PCR (MC Nexus Gradient, Eppendorf AG, Germany) with the temperature profile of initial denaturation of 5 min at 95°C and then 35 cycles at 95°C for 1 min, 53°C for 1 min, and 72°C for 1 min 30 s and 72°C for 8 min for final extension. The amplicons were separated by gel electrophoresis on 1% agarose gels in Bio-Rad gel electrophoresis unit using 1X TAE in buffer tank with electric voltage of 80 V for 1 h.
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
Revolutionizing how scientists
search and build protocols!