ABHD10 cell lysate
Human ABHD10 Baculovirus-Insect cells Overexpression Lysate
Applications
Western Blot
Synonyms
ABHD10; Human ABHD10 Baculovirus-Insect cells Overexpression Lysate; Human ABHD10 (aa 53-306) Insect Cell Lysate (WB positive control); abhydrolase domain containing 10; Human ABHD10 Overexpression Lysate; ABHD10 cell lysate
Host
Baculovirus-Insect 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.
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
271aa
Sequence
Thr53-Asn306
Applicable Applications for ABHD10 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 ABHD10 (Q9NUJ1) (Thr53-Asn306) was fused with a polyhistide tag at the N-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.
Samples are stable for up to twelve months from date of receipt.
Related Product Information for ABHD10 cell lysate
Mycophenolic acid (MPA), the active metabolite of the immunosuppressant mycophenolate mofetil (MMF), is primarily metabolized by glucuronidation to a phenolic glucuronide (MPAG) and an acyl glucuronide (AcMPAG). It is known that AcMPAG, which may be an immunotoxic metabolite, is deglucuronidated in human liver. AcMPAG deglucuronidation activity was detected in both human liver cytosol (HLC) and microsomes (HLM). By purification from HLC with column chromatographic purification steps, the enzyme responsible for AcMPAG deglucuronidationis identified as alpha/beta hydrolase domain containing 1 (ABHD1). Recombinant ABHD1 expressed in Sf9 cells efficiently deglucuronidated AcMPAG with a K(m) value of 1.7 +/- 1.2uM, which was similar to those in HLM, HLC, and human liver homogenates (HLH). Immunoblot analysis revealed ABHD1 protein expression in both HLC and HLM. The AcMPAG deglucuronidation by recombinant ABHD1, HLC, and HLH were potently inhibited by AgNO(3), CdCl(2), CuCl(2), PMSF, bis-p-nitrophenylphosphate, and DTNB. The CL(int) value of AcMPAG formation from MPA, which was catalyzed by human UGT2B7, in HLH was increased by 1.8-fold in the presence of PMSF. Thus, human ABHD1 would affect the formation of AcMPAG, the immunotoxic metabolite.
This Human ABHD10 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of ABHD10 protein from the overexpression lysate was verified.
This Human ABHD10 overexpression lysate was created in Baculovirus-Insect cells and intented for use as a Western blot (WB) positive control. Purification of ABHD10 protein from the overexpression lysate was verified.
References
Nardini M. et al., 1999, Curr Opin Struct Biol. 9 (6): 732-7. Carr PD. et al., 2009, Protein Pept Lett. 16 (10): 1137-48. Cheah E. et al., 1992, Protein Eng. 5 (3): 197-211. Iwamura A. et al., 2012, J Biol Chem. 287 (12): 9240-9.
NCBI and Uniprot Product Information
NCBI GI #
NCBI GeneID
NCBI Accession #
NCBI GenBank Nucleotide #
Molecular Weight
22,141 Da
NCBI Official Full Name
mycophenolic acid acyl-glucuronide esterase, mitochondrial isoform 2
NCBI Official Synonym Full Names
abhydrolase domain containing 10
NCBI Official Symbol
ABHD10
NCBI Protein Information
mycophenolic acid acyl-glucuronide esterase, mitochondrial; abhydrolase domain-containing protein 10, mitochondrial; alpha/beta hydrolase domain-containing protein 10, mitochondrial
UniProt Protein Name
Mycophenolic acid acyl-glucuronide esterase, mitochondrial
Protein Family
UniProt Gene Name
ABHD10
UniProt Synonym Gene Names
Abhydrolase domain-containing protein 10
UniProt Entry Name
ABHDA_HUMAN
NCBI Description
This gene encodes a mitochondrially-localized enzyme that acts in liver cells as a hydrolase. The encoded protein removes glucuronide from mycophenolic acid acyl-glucuronide. There is a pseudogene for this gene on chromosome 6. Alternative splicing results in multiple transcript variants. [provided by RefSeq, Jan 2013]
Uniprot Description
ABHD10: a predicted hydrolase of the alpha/beta hydrolase superfamily
Protein type: EC 3.1.1.93; Protease
Chromosomal Location of Human Ortholog: 3q13.2
Cellular Component: mitochondrion; cytosol
Molecular Function: hydrolase activity, hydrolyzing O-glycosyl compounds
Biological Process: glucuronoside catabolic process
Research Articles on ABHD10
Similar Products
Product Notes
The ABHD10 abhd10 (Catalog #AAA8115380) is a Cell Lysate produced from Baculovirus-Insect cells and is intended for research purposes only. The product is available for immediate purchase. The immunogen sequence is 271aa. AAA Biotech's ABHD10 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 ABHD10 abhd10 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: Thr53-Asn3 06. It is sometimes possible for the material contained within the vial of "ABHD10, 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.Item has been added to Shopping Cart
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