Human carbonic anhydrase XII HEK293 Overexpression Lysate


Human carbonic anhydrase XII HEK293 Overexpression Lysate: 製品の情報

This Human carbonic anhydrase XII overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of carbonic anhydrase XII protein (Cat: 10617-H08H) from the overexpression lysate was verified.
HEK293 Cells
A DNA sequence encoding the extracellular domain of human CA12 (NP_001209.1) (Met 1-Gln 291) was expressed with a C-terminal polyhistidine tag.
The recombinant human CA12 consisting of 278 amino acids and has a calculated molecular mass of 31.6 kDa. As a result of glycosylation, the recombinant protein migrates as an approximately 40-45 kDa protein in SDS-PAGE under reducing conditions.

Human carbonic anhydrase XII HEK293 Overexpression Lysate: 用法

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.
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.
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.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
安定性 & 保存条件
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.
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human carbonic anhydrase XII HEK293 Overexpression Lysate: 別名

Human CA12 Overexpression Lysate; Human CAXII Overexpression Lysate; Human FLJ20151 Overexpression Lysate; Human HsT18816 Overexpression Lysate

carbonic anhydrase XII 背景情報

Carbonic anhydrases (CAs) are a large family of zinc metalloenzymes first discovered in 1933 that catalyze the reversible hydration of carbon dioxide. CAs participate in a variety of biological processes, including respiration, calcification, acid-base balance, bone resorption, and the formation of aqueous humor, cerebrospinal fluid, saliva, and gastric acid. CA12, also known as Car12 and carbonic anhydrase XII, is a type I membrane enzyme of an N-terminal extracellular catalytic domain, a membrane-spanning α-helix, and a small intracellular C-terminal domain. It is highly expressed in colon, kidney, prostate, intestine and activated lymphocytes and moderately expressed in pancreas, ovary, and testis. Overexpression of the CA12 is observed in certain human cancers and is used as a tumor marker. rmCA12 corresponds to the extracellular domain and has both carbonic anhydrase activity and esterase activity.
carbonic anhydrase XII
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