Lysis buffer

Manufactured by Yeasen

Sourced in China

Lysis buffer is a solution used to break open cells and release the contents, including proteins, DNA, and other cellular components. It is a core component in various molecular biology and biochemistry applications that require the extraction and purification of cellular materials.

Automatically generated - may contain errors

Lab products found in correlation

Cell strainer, by BD (1 mentions) Protein a g beads, by Yeasen (1 mentions) Optiprep, by STEMCELL (1 mentions) Protease inhibitor cocktail, by Beyotime (1 mentions) Las 4000epuv mini luminescent image analyzer, by GE Healthcare (1 mentions) Immobilon p membrane, by Merck Group (1 mentions) Enhanced chemiluminescence plus western blotting detection system, by GE Healthcare (1 mentions) Protease inhibitor, by Roche (1 mentions)

7 protocols using lysis buffer

Bone Marrow-Derived Macrophage Generation

Cited 1 time

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

Following the protocol as reported in (Shields et al., 2020 (link)), the BM cells were collected from the femurs with cold phosphate-buffered saline (PBS; Corning) and passed through the cell-strainer (70 μm; BD Falcon), prior to the centrifugation at 1,150 rpm for 5 min at 4°C. Following this, the lysis buffer (Yeasen Biotech, China) was used to remove erythrocytes, which was followed by the centrifugation at 4°C for 5 min at 1,150 rpm. Subsequently, 2 × 10⁵ of cells were cultured each well inside the 24-well plates using DMEM plus 10% fetal bovine serum (FBS), 1% penicillin-streptomycin (P/S) and 10 ng/ml macrophage colony-stimulating factor for 70–72 h. Then, cells were added with fresh medium as mentioned above for 48 h. Subsequently, cells were added with fresh DMEM plus 10% FBS, 1% P/S, lipopolysaccharide (LPS, 100 ng/ml) and interferon-γ (IFN-γ, 20 ng/ml) for 20–24 h, generating BMDM before the experiments as described below.
In addition, L929 and HUVEC were cultured using DMEM plus 10% FBS and 1% P/S and using RPMI-1640 plus 10% FBS and 1% P/S, respectively. In this study, all cells were grown at 37°C with 5% CO₂ and 95% relative humidity.

Wang L., Xia K., Han L., Zhang M., Fan J., Song L., Liao A., Wang W, & Guo J. (2022). Local Administration of Ginkgolide B Using a Hyaluronan-Based Hydrogel Improves Wound Healing in Diabetic Mice. Frontiers in Bioengineering and Biotechnology, 10, 898231.

+ Open protocol

+ Expand

DNA Extraction from Granulosa Cells

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

The DNA in granulosa cells was extracted in lysis buffer (Yeasen Biotech Co., Ltd., Shanghai, China) by heating at 55°C for 30 min, followed by 95°C for 3 min. To extract the DNA present from the original medium, the culture medium was transferred to a microtubes, and cells were removed by centrifugation at 4,000 × g for 1 min, and the spent culture medium was mixed with the same amount of lysis buffer and followed the steps above.

Li X., Wang Z., Wang H., Xu H., Sheng Y, & Lian F. (2022). Role of N-acetylcysteine treatment in women with advanced age undergoing IVF/ICSI cycles: A prospective study. Frontiers in Medicine, 9, 917146.

+ Open protocol

+ Expand

Quantifying Luciferase Activity in Co-cultured HNC Cells

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

The luciferase-expressing HNC cells were co-cultured with fibroblasts and the cell samples were harvested for the detection of luciferase activity 48 h after co-culture. Cells were lysed in 120 μL lysis buffer (Yeasen, China). To measure firefly luciferase activity, 100 μL lysates was mixed with 100 μL of luciferase assay reagent (Yeasen, China), and then the relative luciferase units (RLU) was read with luminometer (Modulus, USA) immediately. The luciferase assay was performed three times in triplicate.

Wang X., Qin X., Yan M., Shi J., Xu Q., Li Z., Yang W., Zhang J, & Chen W. (2019). Loss of exosomal miR-3188 in cancer-associated fibroblasts contributes to HNC progression. Journal of Experimental & Clinical Cancer Research : CR, 38, 151.

+ Open protocol

+ Expand

Immunoprecipitation and Western Blot Analysis

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

Note that, 2.5 μg of mouse anti‐flag, mouse anti‐strep II, rabbit anti‐Rad51, or rabbit anti‐p‐CHK2(Thr68) antibody was first incubated with 30 μl protein‐A/G beads (Yeasen Biotech Co., Shanghai, China) in 250 μl Lysis buffer (with protease inhibitor and phosphatase inhibitor, 1:500, Yeasen) on a rotator at 4°C for 4 h. Meanwhile, 2.5 × 10⁶ transfected 293T cells or two genital ridges were lysed and ultrasonicated in 250 μl IP buffer and then pre‐cleaned with 30 μl protein A/G beads at 4°C for 4 h. Then, the protein A/G‐coupled antibody was incubated with pre‐cleaned lysates at 4°C overnight. Next, after three washes with IP buffer, antibody‐bound beads were subjected to western blotting with corresponding antibodies.

Wang Y., Gao W., Wang L., Wang R., Yang Z., Luo F., He Y., Wang Z., Wang F., Sun Q., Li J, & Zhang D. (2022). FBXW24 controls female meiotic prophase progression by regulating SYCP3 ubiquitination. Clinical and Translational Medicine, 12(7), e891.

+ Open protocol

+ Expand

Iodixanol Gradient Centrifugation for Exosome Isolation

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

To generate a gradient of iodixanol solutions, 50% (w/v), 40% (w/v), and 10% (w/v) iodixanol solutions were prepared by diluting OptiPrep™ (Stemcell™ Technologies) with 0.85% sodium chloride and 10 mmol/L Tris–HCl, pH 7.4. The exosomes were resuspended in 3.8 mL of 50% iodixanol solution, overlaid with 3 mL of 40% solution, and then 2.5 mL of 10% solution in a centrifuge tube. The gradient was then centrifuged at 200,000×g for 2 h at 4 °C. Ten gradient fractions were diluted in PBS and centrifuged at 110,000×g for 70 min at 4 °C. They were then resuspended in a lysis buffer (Yeasen Biotechnology). The density was determined gravimetrically (g/mL).

Yu P., Han Y., Meng L., Tian Y., Jin Z., Luo J., Han C., Xu W., Kong L, & Zhang C. (2024). Exosomes derived from pulmonary metastatic sites enhance osteosarcoma lung metastasis by transferring the miR-194/215 cluster targeting MARCKS. Acta Pharmaceutica Sinica. B, 14(5), 2039-2056.

+ Open protocol

+ Expand

Western Blot Analysis of Apoptosis and Metastasis Markers

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

Proteins were extracted from tissues and cells utilizing Lysis Buffer (Yeasen) containing a protease inhibitor cocktail (Beyotime). An equal amount of protein lysates (30 μg) was separated on 10% or 12% SDS-polyacrylamide gel electrophoresis, and then transferred onto polyvinylidene difluoride membranes. The protein-carrying membranes were incubated with primary antibodies for overnight at 4°C, and with secondary antibodies for 1.5 h at 25°C. GAPDH was chosen as internal control, and gray density of Western blots was analyzed on Image J software (NIH, Bethesda, MD, USA). Relative protein expression levels of B cell lymphoma (Bcl)-2, Bcl-2-associated X protein (Bax), matrix metalloproteinase 2(MMP2) and MMP9 were calculated with normalization to GAPDH. The antibodies are summarized in Table 3.Table 3

The Antibodies Used in This Study

Name	Cat. No.	Dilution Ratio	Source
CHTOP	ab222861	1:5000	Abcam
Bcl-2	ab32124	1:1000	Abcam
Bcl-2	ab196495	1:2000	Abcam
Bax	ab32503	1:10,000	Abcam
Bax	ab3191	1:500	Abcam
MMP2	ab86607	1:500	Abcam
MMP9	ab137867	1:1000	Abcam
MMP9	ab38898	1:1000	Abcam
GAPDH	ab181602	1:10,000	Abcam
Goat anti-Rabbit IgG (HRP)	ab205718	1:50,000	Abcam
Goat anti-mouse IgG (HRP)	ab6789	1:10,000	Abcam

Cao Y., Xie X., Li M, & Gao Y. (2021). CircHIPK2 Contributes to DDP Resistance and Malignant Behaviors of DDP-Resistant Ovarian Cancer Cells Both in vitro and in vivo Through circHIPK2/miR-338-3p/CHTOP ceRNA Pathway. OncoTargets and therapy, 14, 3151-3165.

+ Open protocol

+ Expand

Western Blot Analysis of CAL-62 Cells

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

We used lysis buffer (YEASEN, Shanghai, China) with protease inhibitors (Roche, Indianapolis, IN, USA) to lyse CAL-62 cells. Protein concentrations were determined by the BCA method (Pierce, Therrmo Fisher Scienti c Inc., Rockford, IL, USA). The protein samples were subjected to SDS/PAGE and transferred to PVDF membranes (Immobilon-P membrane, Millipore, Massachusetts, USA). The membranes were blocked with 5% skimmed milk in TBS plus Tween 20 at room temperature for 1 hour, followed by incubation with target antibodies at 4 °C overnight. Information on the antibodies are provided in Table S3. After incubation with HRP-conjugated secondary antibodies for 1 hour, visualization of the protein bands was achieved by an enhanced chemiluminescent chromogenic substrate using the Enhanced Chemiluminescence Plus Western Blotting Detection System (GE Healthcare, Connecticut, USA) and LAS-4000EPUV mini Luminescent Image Analyzer (GE Healthcare).

Ma B., Luo Y., Xu W., Han L., Liu W., Jiang H., Wang X., Wen S., Liao T., Wei W., Ji Q., & Wang Y. (2020). LINC00886, as a Biomarker Indicative of Thyroid Cancer Dedifferentiation, Negatively Regulates the Malignancy in Anaplastic Thyroid Cancer.

+ 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!