Human Carbonic Anhydrase X HEK293 Overexpression Lysate

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Human Carbonic Anhydrase X HEK293 Overexpression Lysate: Product Information

Product Description
This Human Carbonic Anhydrase X overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Carbonic Anhydrase X protein (Cat: 10687-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human CARPX (NP_001076002.1) (Met 1-Asn 300) was expressed with a C-terminal polyhistidine tag.
Molecule Mass
The recombinant human CARPX consists of 291 amino acids and has a predicted molecular mass of 33 kDa. rhCARPX migrates as an approximately 37 kDa band in SDS-PAGE under reducing conditions due to glycosylation.

Human Carbonic Anhydrase X HEK293 Overexpression Lysate: Usage Guide

Preparation Method
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading 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.
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 boil for 2-5 min.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human Carbonic Anhydrase X HEK293 Overexpression Lysate: Alternative Names

Human CA-RPX Overexpression Lysate; Human CA10 Overexpression Lysate; Human CARPX Overexpression Lysate; Human HUCEP-15 Overexpression Lysate

Carbonic Anhydrase X Background Information

Carbonic anhydrase X, also called carbonic anhydrase - related protein X (CARPX) and CA1, belongs to the CA family of zinc metalloenzymes which catalyze the reversible hydration of carbon dioxide in various biological processes such as respiration, renal tubular acidification and bone resorption. The secreted protein CARPX without CA activity (hydration of CO2) is identified as an acatalytic member of the alpha-carbonic anhydrase subgroup. CARP X expression is detected in the adult total brain and almost all parts of the central nervous system, but not in the fetal brain. Accordingly, CARP X is suggested to play a role in the development of central nervous system, especially the brain. The same CARP X protein are encoded by multiple transcript variants of this gene.
Full Name
carbonic anhydrase X
References
  • Okamoto, N. et al., 2001, Biochim. Biophys. Acta 1518:311-316.
  • Taniuchi, K. et al., Neuroscience 112: 93-99.
  • Supuran, C.T. et al., 2003, Med. Res. Rev. 23: 146-189.
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