This Human CALCB overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CALCB protein (Cat: 13283-H05H) from the overexpression lysate was verified.
A DNA sequence encoding the human CALCB (P10092) (Met1-Phe118) was fused with Fc region of mouse IgG at the C-terminus.
The recombinant human CALCB/mFc is a disulfide-linked homodimer. The reduced monomer comprises 327 amino acids and has a predicted molecular mass of 36.2 kDa. The apparent molecular mass of the protein is approximately 37 in SDS-PAGE under reducing conditions.
Human CALCB HEK293 Overexpression Lysate: 使用指南
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.
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).
稳定性 & 储存条件
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 CALCB HEK293 Overexpression Lysate: 别称
Human CALC2 Overexpression Lysate; Human CGRP-II Overexpression Lysate; Human CGRP2 Overexpression Lysate
CALCB, also known as CGPR and calcitonin 2, belongs to the calcitonin family. CALCB is a calcitonin (CT) peptide which may play a role in the mediation of human inflammatory diseases. It is highly expressed in the skin, blood, and cerebrospinal fluid. CGRP immunolabeling (IL) was detected in epidermal keratinocytes at levels that were especially high and widespread in the skin of humans from locations afflicted with postherpetic neuralgia (PHN) and complex region pain syndrome type 1 (CRPS), of monkeys infected with simian immunodeficiency virus, and of rats subjected to L5/L6 spinal nerve ligation, sciatic nerve chronic constriction, and subcutaneous injection of complete Freund's adjuvant. Increased CGRP-IL was also detected in epidermal keratinocytes of transgenic mice with keratin-14 promoter driven overexpression of noggin, an antagonist to BMP-4 signaling. CGPR dilates a variety of vessels including the coronary, cerebral and systemic vasculature.
calcitonin-related polypeptide beta
Lambert NA. 2008, Sci Signal. 1 (25): re5.
Linscheid P. et al., 2005, Endocrinology. 146 (6): 2699-708.