Human FANCA Baculovirus-Insect cells Overexpression Lysate

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Human FANCA Baculovirus-Insect cells Overexpression Lysate: Product Information

Product Description
This Human FANCA overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of FANCA protein (Cat: 11980-H07B) from the overexpression lysate was verified.
Expression Host
Baculovirus-Insect cells
Species
Human
Sequence Information
A DNA sequence encoding the human FANCA (AAH08979.1) (Met 1-Cys 297) was expressed, with a polyhistidine tag at the N-terminus.
Molecule Mass
The recombinant human FANCA consists of 316 amino acids and predicts a molecular mass of 35.2 kDa. It migrates as an approximately 35 kDa band in SDS-PAGE under reducing conditions.

Human FANCA Baculovirus-Insect cells 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 FANCA Baculovirus-Insect cells Overexpression Lysate: Alternative Names

Human FA Overexpression Lysate; Human FA-H Overexpression Lysate; Human FA1 Overexpression Lysate; Human FAA Overexpression Lysate; Human FACA Overexpression Lysate; Human FAH Overexpression Lysate; Human FANCH Overexpression Lysate

FANCA Background Information

FANCA is one of the six known Fanconi anemia gene products (FANCA, FANCC, FANCD2, FANCE, FANCF, and FANCG proteins). Fanconi anemia (FA) is a genetic disorder predisposing to aplastic anemia and cancer characterized by hypersensitivity to DNA-damaging agents and oxidative stress. FANCA associates with the IκB kinase (IKK) signalsome via interaction with IKK2. Components of the FANCA complex undergo rapid, stimulus-dependent changes in phosphorylation, which are blocked by kinase-inactive IKK2.
Full Name
Fanconi anemia, complementation group A
References
  • Otsuki T, et al. (2002) Fanconi anemia protein complex is a novel target of the IKK signalsome. J Cell Biochem. 86 (4): 613-23.
  • Taniguchi T, et al. (2002) The Fanconi anemia protein, FANCE, promotes the nuclear accumulation of FANCC. Blood. 100 (7): 2457-62.
  • Garcia-Higuera I, et al. (1999) Fanconi anemia proteins FANCA, FANCC, and FANCG / XRCC9 interact in a functional nuclear complex. Mol Cell Biol. 19 (7): 4866-73.
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