s2

IMPDH

Protein Name

Helicobacter  pylori inosine-5′-monophosphate dehydrogenase (HpIMPDH)

Tag Name

 

Expression System

E. coli Rosetta-gami™ 2 (DE3)pLysS

Accession Number

 

Product Description

Inosine-5′-monophosphate dehydrogenase (IMPDH) is a key enzyme in the de novo purine biosynthesis pathway, catalyzing the conversion of inosine 5′-monophosphate (IMP) to xanthosine 5′-monophosphate (XMP), a precursor to guanine nucleotides. It is a promising drug target for various diseases, including infections caused by Cryptosporidium parvum, Helicobacter  pylori and Mycobacterium tuberculosis.

Formulation

 

Specific Activity

 

Storage and Stability

20ºC for long term, 4ºC for short term use

Handling Recommendations

Proteins should be stored at cold temperatures when not in use.

To minimise freeze-thaw cycles, aliquot the protein into smaller portions of at least 10 μL to prevent evaporation and maintain consistent concentration.

Quality Control Testing

HpIMPDH Assay

1.11 µg of HpIMPDH reduces NAD+ to NADH in the assay protocol described in page.
2. The specific activity is determined by subtracting the assay controls.

HpIMPDH assay protocol

Chemicals/Materials:
1X Reaction buffer: 50 mM Tris-Cl pH 7.8, 100 mM KCl and 3 mM DTT
2X Substrate buffer: 500 µM IMP and 600 µM NAD+ in 1X reaction buffer
2X Assay buffer: 200 nM HpIMPDH in 1X reaction buffer
Procedure:
1.Mix 100 µL each of substrate buffer and assay buffer in a 96-well black plate to initiate the reaction at 37 °C.
2.Dilute 2X substrate buffer and 2X assay buffers to their 1X concentrations separately in the 1X reaction buffer and use them as controls for the reaction.
3.Measure the specific activity of the enzyme in terms of the fluorescence of NADH (λem = 440 nm and λex = 340 nm).
4.Subtract the reaction reading from the controls to calculate the specific activity of the enzyme.
MS Tryptic Fingerprint: Confirmed identity as HpIMPDH with the translated sequence listed on pages 2 and 3.

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