2X Lysis Buffer (Item J)

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

$10.00  
EL-lysis
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Product Specifications

Size:
5 ml

2X Lysis Buffer for ELISA Kits

2X Lysis Buffer Preparation Instructions
2X Lysis Buffer should be diluted 2-fold with deionized or distilled water. Supplementation with protease or phosphatase inhibitors is recommended. You may use RayBiotech Protease Inhibitor Cocktail (AA-PI), RayBiotech Phosphatase Inhibitor Cocktail Set I (AA-PHI-I) or Phosphatase Inhibitor Cocktail Set II (AA-PHI-II), or the RayBiotech Phosphatase / Protease Inhibitor Set (AA-PIPHI). Alternatively, you can purchase inhibitor cocktails from another manufacturer and follow their specifications.

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

Store at 4°C for 6 months or frozen at -80°C for 1 year
  1. Fang Y., Yao L., Li C., et al. The blockage of the Nogo/NgR signal pathway in microglia alleviates the formation of A? 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
  2. 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
  3. 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
  4. 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:
  5. 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
  6. 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
  7. Miyabayashi K, Ijichi H, Mohri D, et al. Erlotinib prolongs survival in pancreatic cancer by blocking gemcitabine-induced MAPK signals. Cancer Res. 2013 Apr 1;73(7):2221-2234.
    Species: Mouse
    Sample type:
  8. 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
  9. 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
  10. 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

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Usage

This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.

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