Text Size:AAA

Rat COL4A3 Human Cells Transfected Lysate (positive control) (denatured)

DatasheetSpecific ReferencesReviewsRelated ProductsProtocols
COL4A3
Products Description:Human Cells transfected lysate in which Rat COL4A3 has been over-expressed. The whole cell lysate is provided in 1X Sample Buffer.
Host:Rat
Preparation Method:Cell lysate was prepared by homogenization in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined with Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 minutes in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Lysis Buffer:Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF
Quality Control Testing:12.5% SDS-PAGE Stained with Coomassie Blue
Stability:Samples are stable for up to twelve months from date of receipt at -80℃
Recommend Usage:1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.
2. Re-dissolve the pellet using 200μL pure water and boiled for 2-5 min.
3. Store it at -80℃. Recommend to aliquot the cell lysate into smaller quantities for optimal storage. Avoid repeated freeze-thaw cycles.
Notes:The lysate is ready to load on SDS-PAGE for Western blot application. If dissociating conditions are required, add reducing agent prior to heating.
Storage Buffer:In modified RIPA Lysis Buffer
Storage Instruction:Store at -80℃. Aliquot to avoid repeated freezing and thawing
Application notes:WB: Use at an assay dependent dilution.
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Background

COL4A3 is a major structural component of basement membranes. It is composed of 3 alpha subunits, which are encoded by 6 different genes, alpha 1 through alpha 6. Each of these alpha subunits can form a triple helix structure with 2 other subunits to form COL4A3. Autoantibodies bind to the collagen molecules in the basement membranes of alveoli and glomeruli can cause goodpasture syndrome. COL4A3 is also linked to an autosomal recessive form of alport syndrome. COL4A3 is organized in a head-to-head conformation and each gene pair shares a common promoter.

References
  • Ghebrehiwet B. et al., 1992, J Leukoc Biol. 51 (6): 546-56.
  • Maziers N. et al., 2006, Nephrol Ther. 1 (2): 90-100.
  • Rana K. et al., 2005, Semin Nephrol. 25 (3): 163-70.
  • Catalog:80439-R01HL-300
    List Price: $195.00  (Save $0.00)
    Price:$195.00      [How to order]
    Availability2 weeks
    Images
      Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"