Human MFAP3 HEK293 Overexpression Lysate


Human MFAP3 HEK293 Overexpression Lysate: 产品信息

This Human MFAP3 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of MFAP3 protein (Cat: 15313-H02H) from the overexpression lysate was verified.
HEK293 Cells
A DNA sequence encoding the human MFAP3 (NP_005918.1) (Met1-Met147) was expressed with the Fc region of human IgG1 at the C-terminus.
The recombinant human MFAP3 consists 367 amino acids and predicts a molecular mass of 41 kDa.

Human MFAP3 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.
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).
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.

MFAP3 背景信息

MFAP3 (Microfibril Associated Protein 3) is a Protein Coding gene. The human gene encoding MFAP3 has a very simple structure, containing only two translated exons encoding a protein of 362 amino acids. The gene was found to be located on chromosome 5q32-q33.2, near the locus 5q21-q31 reported for the fibrillin gene, FBN2, which has been linked to congenital contractural arachnodactyly. MFAP3 is widely expressed in the placenta, urinary bladder, and other tissues. It does not appear to share homology with any other known protein. MFAP3 is a candidate gene for heritable diseases affecting microfibrils. Diseases associated with MFAP3 include Lutembacher's Syndrome and Postural Orthostatic Tachycardia Syndrome.
microfibrillar-associated protein 3
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