Human FANCA Baculovirus-Insect cells Overexpression Lysate: Product Information
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.
A DNA sequence encoding the human FANCA (AAH08979.1) (Met 1-Cys 297) was expressed, with a polyhistidine tag at the N-terminus.
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
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).
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.
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.
Fanconi anemia, complementation group A
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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