Femto reagent

Manufactured by Thermo Fisher Scientific

Sourced in United States, France

The Femto reagent is a laboratory product manufactured by Thermo Fisher Scientific. It is designed to facilitate specific analytical processes, though its core function is not to be extrapolated upon in this response.

Automatically generated - may contain errors

Lab products found in correlation

Nitrocellulose membrane, by Cytiva (9 mentions) Evolution capt software, by Vilber (6 mentions) Phosphatase inhibitor cocktail 2, by Merck Group (4 mentions) Ne per nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (4 mentions) Fusion solo, by Vilber (3 mentions) Bradford assay kit, by Bio-Rad (3 mentions) Supersignal west pico, by Thermo Fisher Scientific (3 mentions) Skim milk, by BD (2 mentions) Fusion solo chemiluminescence imaging system, by Vilber (2 mentions) Trixon x, by Merck Group (2 mentions) Wet transfer apparatus, by Bio-Rad (2 mentions) Phosphatase inhibitor cocktail, by Merck Group (2 mentions) Ecl grade secondary hrp conjugated antibody, by GE Healthcare (2 mentions) Supersignal pico, by Thermo Fisher Scientific (2 mentions) Protease inhibitor cocktail, by Roche (2 mentions) Superwest signal pico, by Thermo Fisher Scientific (2 mentions) Nitrocellulose membrane, by Bio-Rad (1 mentions) Chemidoc system, by Bio-Rad (1 mentions) Ne per nuclear and cytoplasmic extraction kit, by Thermo Fisher Scientific (1 mentions) Uv 1700 pharmaspec spectrophotometer, by Shimadzu (1 mentions)

18 protocols using femto reagent

Fly Head Protein Extraction and Immunoblotting

Check if the same lab product or an alternative is used in the 5 most similar protocols

Fly heads (100)
were homogenized in RIPA (radioimmunoprecipitation assay) buffer (50
mM Tris-HCl, pH 8.0, 0.5% sodium deoxycholate, 1% Triton X-100, and
150 mM NaCl) containing 1% SDS (sodium dodecyl sulfate) for consecutive
extractions. The protein (30 mg) was separated on a 4–20% gradient
Tris-HCl gel and then transferred to PVDF (polyvinylidene fluoride)
or nitrocellulose membranes (Bio-Rad). The transferred membrane was
then blocked using nonfat milk powder. For blocking nonspecific antibody
binding, the membrane was incubated in TBST buffer (10 mM Tris-HCl,
150 mM NaCl, and 0.1% Tween 20, pH 7.4) containing 5% nonfat milk.
After blocking, primary antibodies against JNK, p-JNK, cleaved caspase
3, and actin were used to probe the membrane (1:1000; Santa Cruz,
USA). Horseradish peroxidase-conjugated secondary antibodies (1:2000)
were utilized for immunodetection. Finally, using a Femto reagent
(Thermo Fisher Scientific, Rockford, USA) on a ChemiDoc system, proteins
were visualized (Bio-Rad, CA, USA).^{22 (link)}

Khatoon R., Kaushik P, & Parvez S. (2022). Mitochondria-Related Apoptosis Regulation by Minocycline: A Study on a Transgenic Drosophila Model of Alzheimer’s Disease. ACS Omega, 7(23), 19106-19112.

+ Open protocol

+ Expand

Protein Extraction and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed by phosphate-buffered saline (PBS) with Triton X-100 (2%), PMSF (1%), and 0.2 mM phosphatase inhibitor cocktail 2 (Sigma-Aldrich, St. Louis, MO, USA) on ice. If necessary, NE-PER Nuclear and Cytoplasmic Extraction Kit (Thermo Fisher Scientific, Waltham, MA, USA) was used to extract cytoplasmic and nuclear protein fractions. Protein concentration was measured by the Bradford method, and 20 μg of protein was resolved by SDS-PAGE, transferred to nitrocellulose membranes (Amersham Biosciences, Piscataway, NJ, USA), and blotted with specific antibodies (Table S3) after blocking with 5% skim milk (Becton, France). The protein bands were visualized with a Femto reagent (Thermo Fisher Scientific, Waltham, MA, USA) and analyzed using Fusion Solo Chemiluminescence Imaging System (Vilber, France) equipped with Evolution Capt software (Vilber, France).

Nguyen M.T, & Lee W. (2022). MiR-320-3p Regulates the Proliferation and Differentiation of Myogenic Progenitor Cells by Modulating Actin Remodeling. International Journal of Molecular Sciences, 23(2), 801.

+ Open protocol

+ Expand

Quantitative Western Blot Analysis of Proteins

Check if the same lab product or an alternative is used in the 5 most similar protocols

Total protein was extracted from C2C12 cells using a lysis buffer containing 0.2 mM PMSF, 2% Triton-X, and 1% phosphatase inhibitor cocktail II (Sigma-Aldrich, St. Louis, MO, USA). Protein concentrations were determined using the Bradford assay with a UV-1700 PharmaSpec spectrophotometer (Shimadzu). The protein samples (20 µg/well) were separated by electrophoresis and transferred onto nitrocellulose membranes (Amersham, Braunschweig, Germany). After blocking with 5% skim milk in TTBS solution (1% Tween 20 in TBS) for 1 h, the membranes were incubated with primary antibodies (Table S3) overnight at 4 °C, washed with TTBS (6 × 5 min), and developed with a secondary antibody (1:10,000 dilution). The blots were then visualized using Femto reagent (Thermo Fisher Scientific, Waltham, MA, USA) and the analytical scanning system Fusion Solo (Vilber, Marne-la-Vallée, France), and the densities of blots were analyzed with Evolution Capt software (Vilber).

Nguyen M.T, & Lee W. (2023). Induction of miR-665-3p Impairs the Differentiation of Myogenic Progenitor Cells by Regulating the TWF1-YAP1 Axis. Cells, 12(8), 1114.

+ Open protocol

+ Expand

Protein Fractionation and Analysis

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

For protein preparation, C2C12 cells were lysed and solubilized with PBS containing 2% Triton-X 100 and 0.1% phosphatase inhibitor cocktail, as previously described [53 (link)]. For nuclear/cytoplasmic protein fractionation, the NE-PER Nuclear and Cytoplasmic Extraction Reagents (Thermo Fisher Scientific, Waltham, MA, USA) were used by following the manufacturer’s protocol. Proteins (20 μg) were resolved by SDS-PAGE and subjected to immunoblot analysis using specific antibodies (Table S3). Blots were visualized using Femto reagent (Thermo Fisher Scientific), detected by Fusion Solo (Vilber, France), and the intensity of immunoblots was analyzed using Evolution Capt software (Vilber, France).

Nguyen M.T., Min K.H, & Lee W. (2021). Palmitic Acid-Induced miR-429-3p Impairs Myoblast Differentiation by Downregulating CFL2. International Journal of Molecular Sciences, 22(20), 10972.

+ Open protocol

+ Expand

Quantitative Immunoblotting of C2C12 Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

C2C12 cells were homogenized using a lysis buffer containing a phosphatase inhibitor cocktail, and lysates were dissolved in Laemmli solution, as previously described [55 (link)]. Total protein concentration was measured with a BSA standard line. Proteins were separated by SDS-PAGE and transferred to nitrocellulose membranes. The membranes were blocked with 5% skim milk (Becton, France) for 1 h. Afterward, immunoblotting was conducted using specific antibodies described in Table 2. All immunoblots were visualized using a Femto reagent (Thermo Fisher Scientific) and quantified by densitometry using an analytical scanning system (Fusion Solo, Paris, France).

Nguyen M.T., Min K.H, & Lee W. (2020). MiR-96-5p Induced by Palmitic Acid Suppresses the Myogenic Differentiation of C2C12 Myoblasts by Targeting FHL1. International Journal of Molecular Sciences, 21(24), 9445.

+ Open protocol

+ Expand

Western Blot Analysis of C2C12 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The C2C12 cells were isolated with lysis buffer consisting of 1% phosphatase inhibitor cocktail II, 2% Trixon-X, and 0.2 mM PMSF (Sigma-Aldrich). All the proteins were quantitated using the Bradford assay with a UV-1700 PharmaSpec spectrophotometer (Shimadzu). The protein samples (20 µg/lane) were separated by electrophoresis and transferred onto nitrocellulose membranes (Amersham). After blocking for 1 h with 5% skimmed milk in TTBS (1% Tween 20 in TBS), the membranes were incubated overnight with specific antibodies (Supplementary Table 3) at 4°C. The next day, the blots were incubated with secondary antibodies (Supplementary Table 3) and developed using chemiluminescent Femto reagent (Thermo Fisher Scientific). Fusion Solo, an analytical scanning system, was used to visualize all blots, and immunoblot intensities were analyzed with Evolution Capt software (Vilber).

Nguyen M.T, & Lee W. (2023). Saturated fatty acid-inducible miR-103-3p impairs the myogenic differentiation of progenitor cells by enhancing cell proliferation through Twinfilin-1/F-actin/YAP1 axis. The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology, 27(3), 277-287.

+ Open protocol

+ Expand

Protein Extraction and Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols

The cells were lysed using lysis buffer (1% phosphatase inhibitor cocktail II, 0.2 mM of PMSF, and 2% Trixon-X in PBS (Sigma-Aldrich, St. Louis, MO, USA)) to obtain the total protein. The lysates were cleared by centrifuging at 13,000 rpm for 5 min at 4 °C. Then, equal amounts of proteins were denatured at 100 °C in an SDS-loading buffer for 10 min. The protein samples (20 µg/well) were subjected to 10% or 8% SDS-PAGE and then were transferred to nitrocellulose membranes (Amersham Biosciences, Piscataway, NJ, USA). The membranes were blocked for 1 h at RT with 5% skim milk in TTBS solution (0.5% TBS-Tween 20), incubated overnight with primary antibodies at 4 °C (Table S4), washed for 6 × 5 min with TTBS, and incubated with a secondary antibody (1:10,000 dilution). Finally, blots were developed using Femto reagent (Thermo Fisher Scientific) and processed using software that measured the densities of protein bands (Fusion Solo, Paris, France).

Nguyen M.T., Ly Q.K., Kim H.J, & Lee W. (2023). WAVE2 Is a Vital Regulator in Myogenic Differentiation of Progenitor Cells through the Mechanosensitive MRTFA–SRF Axis. Cells, 13(1), 9.

+ Open protocol

+ Expand

Protein Extraction and Western Blot Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

The total protein from the transfected cells was obtained using a lysis buffer containing 1% phosphatase inhibitor cocktail II, 0.2 mM PMSF, and 2% Trixon-X (Sigma-Aldrich, St. Louis, MO, USA), and protein concentration was determined using a Bradford assay kit (Bio-Rad, Hercules, CA, USA). Protein samples (20 µg) were resolved on SDS-PAGE gels and transferred to nitrocellulose membranes (Amersham, Braunschweig, Germany), which were blocked with 5% skim milk in TBS-Tween 20 (TTBS) for 1 h and then incubated overnight with the indicated primary antibodies (Table S3) at 4 °C, followed by incubation with secondary antibodies (Table S3) for 1 h at room temperature. Blots were developed using a Femto reagent (Thermo Fisher Scientific, Waltham, MA, USA) with a Fusion Solo Chemiluminescence Imaging System (Vilber Lourmat, Marne-la-Vallée, France), and their densities were analyzed with Evolution Capt software (Vilber Lourmat). β-Actin protein levels were used for normalization.

Nguyen M.T, & Lee W. (2022). Kank1 Is Essential for Myogenic Differentiation by Regulating Actin Remodeling and Cell Proliferation in C2C12 Progenitor Cells. Cells, 11(13), 2030.

+ Open protocol

+ Expand

Western Blot Protein Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed with a lysis buffer (PBS buffer supplemented with 0.2 mM PMSF, 2% Triton-X, and 1% phosphatase inhibitor cocktail II (Sigma-Aldrich, St. Louis, MO, USA)). Protein concentrations were determined using the Bradford assay, and equal amounts of protein (20 µg/well) were resolved via SDS-PAGE and transferred to nitrocellulose membranes. After blocking with 5% skim milk in TTBS solution (0.5% TBS-Tween 20) for 1 h, membranes were incubated overnight at 4 °C with primary antibodies (Table S3). The next day, membranes were washed with TTBS six times and treated with a secondary antibody (1: 10,000 dilution) at room temperature (RT) for 1 h. Proteins were visualized using a chemiluminescent Femto reagent (Thermo Fisher Scientific) and a Fusion Solo S imaging system (Vilber Lourmat, Paris, France). Western blots were quantified using Evolution-Capt v18.10 software supplied by Vilber Lourmat (Paris, France).

Nguyen M.T., Ly Q.K., Kim H.J, & Lee W. (2023). FLII Modulates the Myogenic Differentiation of Progenitor Cells via Actin Remodeling-Mediated YAP1 Regulation. International Journal of Molecular Sciences, 24(18), 14335.

+ Open protocol

+ Expand

Protein Expression Analysis in HepG2 Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

HepG2 cells were lysed in PBS containing 0.2 mM phosphatase inhibitor cocktail 2 (Sigma, Ronkonkoma, NY, USA), 2% Triton X-100, and 1% PMSF. Total protein concentrations were analyzed using the Bradford assay (Bio-Rad). Proteins (20 µg/lane) were resolved by SDS-PAGE and blotted onto nitrocellulose membranes (Amersham Biosciences, Piscataway, NJ, USA), which were blocked with non-fat milk (5%) (Becton, France) for 1 h, and then incubated overnight with specific antibodies at 4 °C (as described in Table S3), washed with TTBS (Tween 20-TBS), and developed with secondary antibodies. Images were obtained using Evolution Capt software (Vilber, France) and commercial Femto reagent (Thermo Fisher Scientific, Waltham, MA, USA). Quantification was performed using Evolution Capt software (Vilber, France).

Nguyen M.T., Min K.H, & Lee W. (2022). MiR-183-5p Induced by Saturated Fatty Acids Hinders Insulin Signaling by Downregulating IRS-1 in Hepatocytes. International Journal of Molecular Sciences, 23(6), 2979.

+ 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?

Revolutionizing how scientists
search and build protocols!