Ecl reagent kit

Manufactured by Beyotime

Sourced in China

The ECL reagent kit is a laboratory product designed for the detection and analysis of proteins in Western blot experiments. It contains the necessary reagents for the enhanced chemiluminescence (ECL) technique, which is used to visualize and quantify target proteins on a membrane. The kit provides the required solutions to generate a luminescent signal that can be detected and measured.

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Lab products found in correlation

Chemiscope 6000 touch imaging system, by Clinx (3 mentions) Sodium salt of isobutyric acid, by Merck Group (2 mentions) Isovaleric acid, by Merck Group (2 mentions) Sodium propionate, by Merck Group (2 mentions) Cd19 pe, by BioLegend (2 mentions) Antibodies to acetyl h3 k9, by ABclonal (2 mentions) Acetyl h3 k18, by ABclonal (2 mentions) Trimethyl h3 k9, by Beyotime (2 mentions) Trimethyl h3 k27, by Beyotime (2 mentions) Easysep human cd3 selection cocktail 2, by STEMCELL (2 mentions) Human cd19 selection cocktail 2, by STEMCELL (2 mentions) Cd2 apc, by BioLegend (2 mentions) Horseradish peroxidase hrp conjugated secondary antibody, by Merck Group (2 mentions) Primescript rt master mix, by Takara Bio (2 mentions) Sodium acetate, by Merck Group (2 mentions) Sodium butyrate, by Merck Group (2 mentions) Gapdh, by Abcam (2 mentions) Sybr green pcr master mix, by Vazyme (2 mentions) Pvdf membrane, by Bio-Rad (1 mentions) β actin, by ABclonal (1 mentions)

5 protocols using ecl reagent kit

Epigenetic Regulation Experimental Protocols

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Sodium acetate (catalog no. S2889), sodium butyrate (catalog no. 303410), sodium propionate (catalog no. P5436), sodium salt of isobutyric acid (catalog no. I1754), and isovaleric acid (catalog no. 129542) were purchased from Sigma-Aldrich. The compounds were used in the form of sodium salt. Antibodies to acetyl-H3-K9 (Abclonal; catalog no. A7255), acetyl-H3-K18 (Abclonal; catalog no. A7257), trimethyl-H3-K9 (Beyotime; catalog no. AF5707), trimethyl-H3-K27 (Beyotime; catalog no. AF5710), and GAPDH (glyceraldehyde-3-phosphate dehydrogenase) (Abcam; catalog no. AB8245) were obtained commercially. PrimeScript RT master mix (TaKaRa; catalog no. RR047A) and SYBR green PCR master mix (catalog no. Q141-02/03; Vazyme, Nanjing, China) were obtained commercially. Lymphocyte separation reagent was purchased from Tianjin Haoyang TBD (catalog no. LTS1077). EasySep human CD3⁺ selection cocktail II (Stem Cell; catalog no. 17851), human CD19⁺ selection cocktail II (Stem Cell; catalog no. 17854), CD2-APC (BioLegend; catalog no. 300311), and CD19-PE (BioLegend; catalog no. 302207) were purchased from commercial sources. Horseradish peroxidase (HRP)-conjugated secondary antibodies were purchased from Sigma-Aldrich. Immunoblots were detected using an ECL reagent kit (Beyotime) and the ChemiScope 6000 Touch imaging system (Clinx, China).

Wang L., Xu D., Huang Q., Yang G., Zhang M., Bi J., Shan J., Li E, & He S. (2021). Characterization of Tonsil Microbiota and Their Effect on Adenovirus Reactivation in Tonsillectomy Samples. Microbiology Spectrum, 9(2), e01246-21.

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Epigenetic Regulation Experimental Protocols

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Formononetin's Impact on Endometrial Cancer Cells

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After 48 h of incubation with the designated concentration of formononetin, the Ishikawa, HEC-1A, and HEC-251 cells were lysed in RIPA buffer. The quantified protein was separated by 10% SDS-PAGE and subsequently transferred to polyvinylidene difluoride (PVDF) membranes (Bio-Rad, USA). Then the membrane was sealed with 5% nonfat milk and incubated with primary antibodies including ERβ (1:2500; Proteintech Group, Inc. Wuhan, China), p53 (1:800; bsm-33058M; Beijing Bioss Biological Co., Ltd, Beijing, China), and β-actin (ABclonal, Inc. Wuhan, China) for 2 h at room temperature. After three washes with the TBST solution, the samples were incubated with the appropriate secondary antibodies for 2 h. The blots were visualized with an electro-chemiluminescence (ECL) reagent kit (Beyotime, China). Image J software (National Institutes of Health, Bethesda, MD, USA) was used to quantify each protein band intensity, and the β-actin was used as an internal reference.

Zhang Q, & Huang X. (2021). The modulatory properties of Astragalus membranaceus treatment on endometrial cancer: an integrated pharmacological method. PeerJ, 9, e11995.

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Western Blot Analysis of LINE-1 and TASOR

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Live cells were lysed in protein extraction buffer [50 mM Tris pH 7.4, 150 mM NaCl, 1% Triton X-100, 1% sodium deoxycholate, 0.1% sodium dodecylsulfate (SDS), 10 μM EDTA] with protease inhibitors for 30 min at 4°C. After a brief centrifugation, the supernatant of the cell lysate was collected and separated by 8% SDS–polyacrylamide gel electrophoresis (PAGE). Proteins were transferred to polyvinylidene difluoride (PVDF) membranes. Membranes were blocked with 2% non-fat milk in TBST (Tris-buffered saline; 100 mM Tris–HCl, pH 7.4, 150 mM NaCl, 0.1% Tween-20) followed by incubation with rabbit anti-LINE-1 ORF1p antibody (ab230966, Abcam) or rabbit anti-FAM208 (TASOR) antibody (ab224393, Abcam) at 4°C overnight. Membranes were washed three times with TBST and then incubated with a horseradish peroxidase (HRP)-conjugated mouse anti-rabbit secondary antibody (31464, Thermofisher Scientific) for 30 min at room temperature. After washing three times with TBST, proteins on the PVDF membrane were detected using an ECL reagent kit (P0018FS, Beyotime, Shanghai, China) and the ChemiScope 6000 Touch imaging system (Clinx, China). GAPDH was used as a loading control which was detected with mouse anti-GAPDH antibody (AF2819, Beyotime) followed by an HRP-conjugated rabbit anti-mouse secondary antibody (61–6520, Thermofisher Scientific).

Zhang M., Sun W., You X., Xu D., Wang L., Yang J., Li E, & He S. (2023). LINE-1 repression in Epstein–Barr virus-associated gastric cancer through viral–host genome interaction. Nucleic Acids Research, 51(10), 4867-4880.

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Western Blot Analysis of Protein Markers

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Protein samples of MLE-12 were segregated by using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and then transferred on nitrocellulose membranes. Membranes were blocked in 5% dry milk, and incubated with primary antibodies: anti-EDA2R and anti-βactin (1:2000), anti-Bax and anti-Bcl-2 (1:3000), anti-p-IκBα (p-nuclear factor of kappa light polypeptide gene enhancer in B-cells inhibitor, alpha) and anti-IκBα (1:4000), and antip-p65 and anti-p65 (1:5000). The membranes were incubated with secondary antibodies (1:5000), and subjected to electrochemiluminescence (ECL) reagent kit (Beyotime). All antibodies were purchased from Abcam.

Jia N., Jia Y., Yang F, & Du W. (2022). Knockdown of EDA2R alleviates hyperoxia-induced lung epithelial cell injury by inhibiting NF-κB pathway. Allergologia et immunopathologia, 50(5).

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