2μg (R: reducing condition, N:non-reducing condition).
PAPP-A His Tag Protein, Human
PAPP-A His Tag Protein, Human
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Product Details
Product Details
Product Specification
| Species | Human |
| Synonyms | Pappalysin-1, PAPPA |
| Accession | Q13219 |
| Amino Acid Sequence | Glu82-Gly1627 with His Tag at the C-Terminus |
| Expression System | HEK293 |
| Molecular Weight | 50-55kDa&170-180kDa&＞180kDa (Reducing) |
| Purity | ＞95% by SDS-PAGE |
| Conjugation | Unconjugated |
| Tag | His Tag |
| Physical Appearance | Lyophilized powder |
| Storage Buffer | PBS, pH7.4, 5% trehalose |
| Reconstitution | Reconstitute at 0.1-1 mg/ml according to the size in ultrapure water after rapid centrifugation. |
| Stability & Storage |
· 12 months from date of receipt, lyophilized powder stored at -20 to -80℃. · 3 months, -20 to -80℃ under sterile conditions after reconstitution. · 1 week, 2 to 8℃ under sterile conditions after reconstitution. · Please avoid repeated freeze-thaw cycles. |
| Reference | 1.Lawrence JB, Oxvig C, Overgaard MT, Sottrup-Jensen L, Gleich GJ, Hays LG, Yates JR 3rd, Conover CA. The insulin-like growth factor (IGF)-dependent IGF binding protein-4 protease secreted by human fibroblasts is pregnancy-associated plasma protein-A. Proc Natl Acad Sci U S A. 1999 Mar 16;96(6):3149-53. |
Background
PAPP-A (Pregnancy-Associated Plasma Protein-A) is a zinc metalloproteinase comprising multiple functional modules including LNR1, LNR2, M1-M5 metalloprotease domains, and CPD (CUB and protease-like domain); its activity is covalently regulated by endogenous inhibitors proMBP and STC2 through steric hindrance mechanisms, and cryo-EM structural studies published in Cell Discovery and Nature Communications in 2024 first resolved the three-dimensional structures of these complexes, laying the foundation for targeted drug design; functionally, PAPP-A specifically cleaves insulin-like growth factor binding proteins (IGFBP-2, -4, -5) to release active IGF-I/II, serving as a key regulator of the IGF signaling pathway involved in fetal growth and development, bone formation, tissue repair, and metabolic regulation; clinically, PAPP-A is a core serum biomarker for first-trimester Down syndrome screening (low levels indicating fetal aneuploidy risk), with low PAPP-A also associated with fetal growth restriction, preeclampsia, and preterm delivery, while elevated levels are important biomarkers for atherosclerotic plaque instability and acute coronary syndrome, and its overexpression in ovarian cancer, breast cancer, Ewing sarcoma, and other malignancies promotes tumor progression, making it a potential target for cardiovascular risk assessment and antitumor therapy.
Protocol
Assay protocol
Principle: As an active protease, recombinant human PAPP-A cleaves IGFBP-5 under the conditions described in Activity Assay Protocol.
Materials
1.Assay Buffer: 50 mM Tris, 10 mM CaCl2, 150 mM NaCl (TCN)
2.PAPP-A His Tag Protein, Human
3.Human IGFBP5 Protein, His Tag (S0A0185)
4.4-20% SDS-PAGE gel
5.5×reducing SDS-PAGE gel buffer
6.SDS-PAGE
Produce
1. Dilute PAPP-A to 10 µg/mL, in Assay Buffer.
2. Dilute rhIGFBP-5 to 200 µg/mL in Assay Buffer
3. Combine equal volumes of 10 µg/mL PAPP-A and 200 µg/mL rhIGFBP-5. Include two blank controls containing Assay Buffer in place of PAPP-A.
4. Incubate reaction vials and one control at 37 °C for 30 minutes. Keep the other control at -20 °C during the incubation period.
5. Combine 20 µL from each reaction vial with 5µL 5×reducing SDS-PAGE gel buffer.
6. Analyze the cleavage by SDS PAGE (load 20 µL/lane) followed by protein staining.
Picture
Picture
SDS-PAGE
