ACP5 Protein, Human (HEK293, His)

The ACP5 protein actively dephosphorylates osteopontin and bone sialoprotein. Its upregulated expression in pathological conditions like Gaucher and Hodgkin diseases, hairy cell, B-cell, and T-cell leukemias suggests a potential association with these diseases. This underscores ACP5's relevance in diverse pathological states, indicating its role in modulating pathways related to bone health and hematopoietic system disorders. ACP5 Protein, Human (HEK293, His) is the recombinant human-derived ACP5 protein, expressed by HEK293 , with C-His labeled tag.

The ACP5 protein is actively engaged in the dephosphorylation of osteopontin and bone sialoprotein. Notably, its expression appears to be upregulated in specific pathological conditions, including Gaucher and Hodgkin diseases, as well as hairy cell, B-cell, and T-cell leukemias. This suggests a potential association between ACP5 expression and these pathological states, indicating its relevance in the context of diverse diseases. The involvement of ACP5 in the dephosphorylation of key proteins associated with bone metabolism highlights its role in modulating pathways related to bone health and pathological conditions affecting the hematopoietic system.

Measured by its ability to cleave a substrate, p-Nitrophenyl phosphate (pNPP). The specific activity is 37780 pmoL/min/μg.

Materials
Assay buffer: 50mM NaOAc, pH 5.0
ACP5 Protein, Human (HEK293, His) (HY-P75561)
Substrate:p-nitrophenylphosphate, 200mM stock solution with DMSO
NaOH, 0.2 M solution prepared with deionized water

Procedure
1. Dilute Human ACP5 to 0.1 μg/mL with experimental buffer.
2. Dilute the substrate to 2 mM with buffer.
3. Add 50 μL of Human ACP5 and 50 μL of 2 mM substrate to a 96-well plate. Additionally, set up blank controls containing 50 μL of assay buffer and 50 μL of 2 mM substrate. Each concentration is performed in triplicate wells.
4. Incubate the reaction at room temperature for 10 minutes, protected from light.
5. Add 100 μL of 0.2 M NaOH to each well to terminate the reaction and develop the color.
6. Preparation of Standard Solution: First, prepare a 1 M p-nitrophenol stock solution. Then dilute the stock solution to concentrations of 12.5, 6.25, 7.125, 1.56, 0.78, 0.39, and 0 μM respectively, with each concentration tested in triplicate wells.
7. Read the OD value of each well at 410 nm endpoint mode.
8. Calculate the specific activity

Human

HEK293

C-His

P13686 (A22-P320)

54  [NCBI]

Partial

ATPALRFVAVGDWGGVPNAPFHTAREMANAKEIARTVQILGADFILSLGDNFYFTGVQDINDKRFQETFEDVFSDRSLRKVPWYVLAGNHDHLGNVSAQIAYSKISKRWNFPSPFYRLHFKIPQTNVSVAIFMLDTVTLCGNSDDFLSQQPERPRDVKLARTQLSWLKKQLAAAREDYVLVAGHYPVWSIAEHGPTHCLVKQLRPLLATYGVTAYLCGHDHNLQYLQDENGVGYVLSGAGNFMDPSKRHQRKVPNGYLRFHYGTEDSLGGFAYVEISSKEMTVTYIEASGKSLFKTRLP

Approximately 35 kDa

• 
  Greater than 95% as determined by reducing SDS-PAGE.

Lyophilized powder.

Lyophilized from a 0.22 μm filtered solution of PBS, pH 7.4.

<1 EU/μg, determined by LAL method.

It is not recommended to reconstitute to a concentration less than 100 μg/mL in ddH₂O. For long term storage it is recommended to add a carrier protein (0.1% BSA, 5% HSA, 10% FBS or 5% Trehalose).

Stored at -20°C for 2 years from date of receipt. After reconstitution, it is stable at 4°C for 1 week or -20°C for longer (with carrier protein). It is recommended to freeze aliquots at -20°C or -80°C for extended storage.

Room temperature in continental US; may vary elsewhere.