2X Lysis Buffer

2X Lysis Buffer for Antibody Array Kits

$15.00
SDS
AA-LYS-10ml
+

Product Specifications

Size:
10 ml

2X Lysis Buffer for Antibody Array 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
SDS
  1. Che G., Wang Y., Zhou B., et al. Knockdown of Heparanase Suppresses Invasion of Human Trophoblasts by Activating p38 MAPK Signaling Pathway. Dis Markers. 2018 Apr 17;2018:7413027. doi: 10.1155/2018/7413027.
    Species: Human
    Sample type: Cell Lysate
  2. Wang X., Zhang M., Ping F., et al. Identification and therapeutic intervention of coactivated anaplastic lymphoma kinase, fibroblast growth factor receptor 2, and ephrin type-A receptor 5 kinases in hepatocellular carcinoma. Hepatology. 2018 Jan 21. doi: 10.1002/hep.29792.
    Species: Human
    Sample type: Cell Lysate
  3. Lin J., Peng Y., Wang S., et al. Sympathetic Nervous System Control of Carbon Tetrachloride-Induced Oxidative Stress in Liver through ?-Adrenergic Signaling. Oxid Med Cell Longev. 2016;2016:3190617. doi: 10.1155/2016/3190617
    Species: Mouse
    Sample type: Tissue Lysate
  4. Ge H., Zhu H., Xu N., et al. Increased Lung Ischemia-Reperfusion Injury in Aquaporin 1-Null Mice Is Mediated via Decreased Hypoxia-Inducible Factor 2? Stability. Am J Respir Cell Mol Biol. 2016 Jun;54(6):882-91. doi: 10.1165/rcmb.2014-0363OC.
    Species: Mouse
    Sample type: NA
  5. Qin Y., Zhang S., Den S., et al. Epigenetic silencing of miR-137 induces drug resistance and chromosomal instability by targeting AURKA in multiple myeloma. Leukemia. 2017 Jan 6. doi: 10.1038/leu.2016.325.
    Species: Human
    Sample type: Cell Lysate
  6. Chen J, Liu Z, Hong MM, Zhang H, Chen C, et al. (2014) Proangiogenic Compositions of Microvesicles Derived from Human Umbilical Cord Mesenchymal Stem Cells. PLoS ONE 9(12): e115316. doi:10.1371/journal.pone.0115316
    Species: NA
    Sample type: N/A
  7. Hawkins JL., Denson JE., Miley DR., Durham PL. Nicotine stimulates expression of proteins implicated in peripheral and central sensitization. Neuroscience. 2015 Apr 2;290:115-25. doi: 10.1016/j.neuroscience.2015.01.034.
    Species: Rat
    Sample type: Tissue Lysate
  8. Zychowska M. Rojewska E., Pilat D., Mika J. The Role of Some Chemokines from the CXC Subfamily in a Mouse Model of Diabetic Neuropathy. J Diabetes Res. 2015;2015:750182. doi: 10.1155/2015/750182.
    Species: Mouse
    Sample type: Tissue Lysate
  9. Li HWR., Lioa S., Chiu P., et al. Effects of adrenomedullin on the expression of inflammatory cytokines and chemokines in oviducts from women with tubal ectopic pregnancy: an in-vitro experimental study. Reprod Biol Endocrinol. 2015; 13: 120.doi: 10.1186/s12958-015-0117-x
    Species: Human
    Sample type: Tissue Lysate
  10. Zhang W., Liu N., Shi H., et al. Upregulation of BMSCs Osteogenesis by Positively-Charged Tertiary Amines on Polymeric Implants via Charge/iNOS Signaling Pathway. Sci Rep. 2015 Mar 20;5:9369. doi: 10.1038/srep09369.
    Species: N/A
    Sample type: N/A

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Usage

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

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