IMPAD1, Recombinant, Human, aa34-359, His-Tag (Inositol Monophosphatase 3)

Size

£512.00

SKU:: 218267
Molecular Weight:: 37.6
Purity:: ~90% (SDS-PAGE)
Storage Conditions:: -20°C
Supplier:: United States Biological
Format:: Supplied as a liquid in PBS, pH 7.4. No preservative added.
Extra Details:: IMPAD1, also known as Inositol monophosphatase 3, is a member of the inositol monophosphatase family. IMPAD1 is localized to the Golgi apparatus and catalyzes the hydrolysis of phosphoadenosine phosphate (PAP) to adenosine monophosphate (AMP). Mutations in this gene are a cause of GRAPP type chondrodysplasia with joint dislocations, and a pseudogene of this gene is located on the long arm of chromosome 1. Recombinant human IMPAD1 protein was purified by conventional chromatography after refolding of the isolated inclusion bodies in a renaturation buffer. Source: Recombinant protein corresponding to aa34-359 from human IMPAD1, fused to His-tag at N-terminus, expressed in E.coli. Molecular Weight: ~37.6kD (349aa) Applications: Suitable for use in SDS-PAGE. Other applications not tested. Recommended Dilution: Optimal dilutions to be determined by the researcher. Specific Activity: ~3300pmole/min/ug, determined by its ability to dephosphorylate adenosine 3'5'-diphosphate sodium slat at pH 7.5, 25°C. Activity Assay: 1. Prepare 1mM PAP in 20mM Tris-HCl, pH 7.5, 15mM MgCl2. 2. Dilute IMPAD1 in various concentrations (20ug/ml, 10ug/ml, 5ug/ml) in 20mM Tris-HCl, pH 7.5, 15mM MgCl2, containing blank of 20mM Tris-HCl, 15mM MgCl2. 3. Make the phosphate standard range of 0.31-20nmol/well. 4. Load 25ul of 1mM PAP, add 25ul of diluted IMPAD1 to well, and load 50ul of phosphate standard. 5. Incubate for 15 minute at 25°C. 6. Add 150ul of Malachite green solution mix and incubate for 15 minutes at 25°C. 7. Read at 650nm. - Malachite green solution: ammonium molybdate 675mg, Malachite green carbinol hydrochloride, 3.3mg in 50ml of 1M H2SO4. Storage and Stability: May be stored at 4°C for short-term only. Aliquot to avoid repeated freezing and thawing.. Store at -20°C. Aliquots are stable for 6 months after receipt at -20°C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.

Shipping Conditions:: Blue Ice