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GLA/alpha-Galactosidase A Cell Lysate | GLA cell lysate

Human GLA/alpha-Galactosidase A HEK293 Overexpression Lysate

Gene Names
GLA; GALA
Applications
Western Blot
Synonyms
GLA/alpha-Galactosidase A; Human GLA/alpha-Galactosidase A HEK293 Overexpression Lysate; Human alpha-Galactosidase A/GLA HEK293 Cell Lysate (WB positive control); galactosidase; alpha; Human GALA Overexpression Lysate; GLA cell lysate
Ordering
For Research Use Only!
Host
HEK293 Cells
Form/Format
1x Sample Buffer (1x modified RIPA buffer+1x SDS loading 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.
Sequence Positions
409aa
Sequence
Met1-Leu429
Applicable Applications for GLA cell lysate
Western Blot (WB)
Application Notes
WB: Use at an assay dependent dilution.
Species
Human
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 200uL pure water and boil for 2-5 min.
Sequence Construction
A DNA sequence encoding the human GLA (NP_000160.1) (Met 1-Leu 429) was fused with a polyhistidine tag at the C-terminus.
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. Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
Preparation and Storage
Store at 4 degree C for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80 degree C for up to twelve months. Avoid repeated freeze-thaw cycles.
Samples are stable for up to twelve months from date of receipt.
Related Product Information for GLA cell lysate
Alpha-galactosidase A, also known as Alpha-D-galactoside galactohydrolase, Alpha-D-galactosidase A, Melibiase and GLA, is a member of the glycosyl hydrolase 27 family. GLA is used as a long-term enzyme replacement therapy in patients with a confirmed identification of Fabry disease. Defects in GLA are the cause of Fabry disease (FD) which is a rare X-linked sphingolipidosis disease where glycolipid accumulates in many tissues. The disease consists of an inborn error of glycosphingolipid catabolism. FD patients show systemic accumulation of globotriaoslyceramide (Gb3) and related glycosphingolipids in the plasma and cellular lysosomes throughout the body. Clinical recognition in males results from characteristic skin lesions (angiokeratomas) over the lower trunk. Patients may show ocular deposits, febrile episodes, and burning pain in the extremities. Death results from renal failure, cardiac or cerebral complications of hypertension or other vascular disease. Deficiency of GLA leads to the accumulation of glycosphingolipids in the vasculature leading to multiorgan pathology. In addition to well-described microvascular disease, deficiency of GLA is also characterized by premature macrovascular events such as stroke and possibly myocardial infarction.

This Human GLA/alpha-Galactosidase A overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of GLA/alpha-Galactosidase A protein from the overexpression lysate was verified.
References
Koide T.et al., 1990, FEBS Lett. 259:353-356. Yang C.-C. et al., 2003, Clin. Genet. 63:205-209. Verovnik F. et al., 2004, Eur. J. Hum. Genet. 12:678-681. Nance C.S. et al., 2006, Arch. Neurol. 63:453-457.

NCBI and Uniprot Product Information

NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
UniProt Accession #
Molecular Weight
48,767 Da
NCBI Official Full Name
alpha-galactosidase A
NCBI Official Synonym Full Names
galactosidase, alpha
NCBI Official Symbol
GLA
NCBI Official Synonym Symbols
GALA
NCBI Protein Information
alpha-galactosidase A; melibiase; alpha-gal A; agalsidase alfa; alpha-D-galactosidase A; alpha-D-galactoside galactohydrolase 1
UniProt Protein Name
Alpha-galactosidase A
UniProt Gene Name
GLA
UniProt Entry Name
AGAL_HUMAN

NCBI Description

This gene encodes a homodimeric glycoprotein that hydrolyses the terminal alpha-galactosyl moieties from glycolipids and glycoproteins. This enzyme predominantly hydrolyzes ceramide trihexoside, and it can catalyze the hydrolysis of melibiose into galactose and glucose. A variety of mutations in this gene affect the synthesis, processing, and stability of this enzyme, which causes Fabry disease, a rare lysosomal storage disorder that results from a failure to catabolize alpha-D-galactosyl glycolipid moieties. [provided by RefSeq, Jul 2008]

Uniprot Description

GLA: Defects in GLA are the cause of Fabry disease (FD). FD is a rare X-linked sphingolipidosis disease where glycolipid accumulates in many tissues. The disease consists of an inborn error of glycosphingolipid catabolism. FD patients show systemic accumulation of globotriaoslyceramide (Gb3) and related glycosphingolipids in the plasma and cellular lysosomes throughout the body. Clinical recognition in males results from characteristic skin lesions (angiokeratomas) over the lower trunk. Patients may show ocular deposits, febrile episodes, and burning pain in the extremities. Death results from renal failure, cardiac or cerebral complications of hypertension or other vascular disease. Heterozygous females may exhibit the disorder in an attenuated form, they are more likely to show corneal opacities. Belongs to the glycosyl hydrolase 27 family.

Protein type: Carbohydrate Metabolism - galactose; Lipid Metabolism - sphingolipid; Glycan Metabolism - glycosphingolipid biosynthesis - globo series; Hydrolase; EC 3.2.1.22; Lipid Metabolism - glycerolipid

Chromosomal Location of Human Ortholog: Xq22

Cellular Component: Golgi apparatus; lysosomal lumen; lysosome; cytoplasm; extracellular region

Molecular Function: protein binding; protein homodimerization activity; hydrolase activity; alpha-galactosidase activity; galactoside binding; catalytic activity; receptor binding

Biological Process: sphingolipid metabolic process; negative regulation of nitric-oxide synthase activity; glycoside catabolic process; negative regulation of nitric oxide biosynthetic process; glycosphingolipid catabolic process; glycosylceramide catabolic process; glycosphingolipid metabolic process; oligosaccharide metabolic process

Disease: Fabry Disease

Research Articles on GLA

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Product Notes

The GLA gla (Catalog #AAA8114882) is a Cell Lysate produced from HEK293 Cells and is intended for research purposes only. The product is available for immediate purchase. The immunogen sequence is 409aa. AAA Biotech's GLA/alpha-Galactosidase A can be used in a range of immunoassay formats including, but not limited to, Western Blot (WB). WB: Use at an assay dependent dilution. Researchers should empirically determine the suitability of the GLA gla for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. The amino acid sequence is listed below: Met1-Leu42 9. It is sometimes possible for the material contained within the vial of "GLA/alpha-Galactosidase A, Cell Lysate" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.

Precautions

All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.

Disclaimer

Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.

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