2X Lysis Buffer (Item J)

2X Cell Lysis Buffer for use with ELISA Kits (5 ml)

Specifications

Size: 5 ml

Method Of Detection: Colorimetric

Lysate Preparation

Cell Lysates:

Rinse cells with PBS, taking care to remove any remaining PBS before detaching cells. Detach  2 x 10⁷ cells by trypsinization or scraping and transfer the cells to a microfuge tube. Centrifuge to pellet the cells and then remove any remaining buffer. Add ~500 µl prepared lysis buffer and pipette up and down to resuspend the pellet. Incubate the lysates with shaking at 4° C for 30 minutes. Spin down the tubes in a microfuge at top speed for 10 minutes at (4°C), and transfer the supernatants to a clean tube.

Tissue Lysates:

Transfer approximately 100 mg crude tissue into a tube with 1 ml 1X Lysis Buffer. Homogenize the tissue according to homogenizer manufacturer instructions. Transfer extracts to microfuge tubes and centrifuge at top speed for 20 minutes (4°C).

Lysate Application

Cell or tissue lysates should be used immediately or aliquoted and stored at –80°C. Avoid repeated freeze-thaw cycles. Thawed lysates should be kept on ice prior to use.

For the first experiment, we recommend serial dilution testing, starting with a 5-fold dilution, to determine the optimal protein load for the assay. Optimal dilution depends on the abundance of target proteins and should be determined empirically,

The total protein concentration may be determined using the BCA Protein Assay Kit from Pierce, Cat#: 23227. We recommended to dilute your lysate to a final total protein concentration of 50-500 ug/ml (5-fold or more is preferred) when performing antibody array or ELISA testing.

Storage

Storage at 4°C for 6 months or frozen at -80°C for 1 year

Catalog #: EL-lysis

Please email us at cs@medikonia.com for any enquiry. To place an order, please include the catalog number(s) of the product(s) in the email.

Safety Data Sheet

Citations

Fang Y., Yao L., Li C., et al. The blockage of the Nogo/NgR signal pathway in microglia alleviates the formation of Aβ plaques and tau phosphorylation in APP/PS1 transgenic mice. J Neuroinflammation. 2016; 13: 56. Published online 2016 Mar 3. doi: 10.1186/s12974-016-0522-x

Species: Mouse

Sample Type: Tissue lysate (brain tissue lysates in Ab plaques)

Wu S-Y, Yu J-S, Liu F-T, et al. Galectin-3 negatively regulates dendritic cell production of IL-23/IL-17-axis cytokines in infection by Histoplasma capsulatum. J. Immunol. 2013 Apr 1;190(7):3427-3437.
Species: Mouse
Sample Type: Tissue Lysate (Spleen Lysate)

Lee, Agnes Fermin, et al. "Galectin-3 modulates Th17 responses by regulating dendritic cell cytokines." The American journal of pathology 183.4 (2013): 1209-1222.
Species: Mouse
Sample Type: Tissue Lysate (brain and kidney)

Zhang Q., Fong C., Yu W., et al. Herbal formula Astragali Radix and Rehmanniae Radix exerted wound healing effect on human skin fibroblast cell line Hs27 via the activation of transformation growth factor (TGF-?) pathway and promoting extracellular matrix (ECM) deposition. Phytomedicine. 2012 Dec 15;20(1):9-16. doi: 10.1016/j.phymed.2012.09.006
Species: Human
Sample Type: Cell Lysate (Hs27 human fibroblast cell lines)

Sen S., Barb J., Cherukuri P., Accame D., Elkahloun A., Singh L., et al. Identification of candidate genes involved in coronary artery calcification by transcriptome sequencing of cell lines. BMC Genomics 2014, 15:198 doi:10.1186/1471-2164-15-198
Species: N/A
Sample Type: (Human Carotid atherosclerotic plaques)

Qin M., Luo Y., Meng X., et al. Myricitrin attenuates endothelial cell apoptosis to prevent atherosclerosis: An insight into PI3K/Akt activation and STAT3 signaling pathways. Vascul Pharmacol. 2015 Jul;70:23-34. doi: 10.1016/j.vph.2015.03.002
Species: Human
Sample Type: Cell Lysate (HUVEC Cells)

Zhu, Min, et al. "Comparison of three different fat graft preparation methods: gravity separation, centrifugation, and simultaneous washing with filtration in a closed system." Plastic and reconstructive surgery 131.4 (2013): 873-880.
Species: Human
Sample Type: Tissue Lysate (fat grafts)

Sidhu P., et al. Anticancer activity of VDR-coregulator inhibitor PS121912. Cancer Chemotherapy and Pharmacology Published Online August 2014. DOI 10.1007/s00280-014-2549-y
Species: Human
Sample Type: N/A

Chao Yang, Deqiang Lei, Weixiang Ouyang, et al., “Conditioned Media from Human Adipose Tissue-Derived Mesenchymal Stem Cells and Umbilical Cord-Derived Mesenchymal Stem Cells Efficiently Induced the Apoptosis and Differentiation in Human Glioma Cell Lines In Vitro,” BioMed Research International, vol. 2014, Article ID 109389, 13 pages, 2014. doi:10.1155/2014/109389

Species: Human

Sample Type: Conditioned Media (Adipose-derived Stem Cell and Umbilical derived stem c)

Li R.L., et al. Postoperative impairment of cognitive function in old mice: a possible role for neuroinflammation mediated by HMGB1, S100B, and RAGE. Journal of Surgical Research, 185(2013) 815 -824. http://dx.doi.org/10.1016/j.jss.2013.06.043

Species: Mouse

Sample Type: Tissue Lysate (Hippocampal lysate)