Recombinant Rat HTRA1 Protein

• Cat.No.: HTRA1-2972R
• Product Overview: Recombinant Rat HTRA1 full length or partial length protein was expressed.

Specification

• Species: Rat
• Source: Mammalian Cells
• Tag: His
• Form: Liquid or lyophilized powder
• Endotoxin: < 1.0 EU per μg of the protein as determined by the LAL method.
• Purity: >80%
• Notes: This item requires custom production and lead time is between 5-9 weeks. We can custom produce according to your specifications.
• Storage: Store it at +4 ºC for short term. For long term storage, store it at -20 ºC~-80 ºC.
• Storage Buffer: PBS buffer

Gene Information

• Gene Name: Htra1 HtrA serine peptidase 1 [ Rattus norvegicus ]
• Official Symbol: HTRA1
• Gene ID: 65164
• mRNA Refseq: NM_031721.1
• Protein Refseq: NP_113909.1
• UniProt ID: Q9QZK5

Citation

HtrA3 as an Early Marker for Preeclampsia: Specific Monoclonal Antibodies and Sensitive High-Throughput Assays for Serum Screening

Journal: PLoS ONE    PubMed ID: 23049902    Data: 2022/11/28

Authors: Kemperly Dynon, Sophea Heng, Haibin Wang

Article Snippet:C-terminally His-tagged full length HtrA1 protein (produced in insect cells) and mature HtrA2 protein (produced in Escherichia coli) were purchased from Creative BioMart (Shirley, NY, USA).. C-terminally His-tagged human wild type (WT) HtrA3-L produced in insect cells was purchased from ProteaImmun GmbH (Berlin, Germany).C-terminally His-tagged human wild type (WT) HtrA3-L produced in insect cells was purchased from ProteaImmun GmbH (Berlin, Germany).

( A ) An equal amount (50 ng) of recombinant (r) human HtrA proteins [rHtrA1 (A1), rHtrA2 (A2) and rHtrA3 (A3, HtrA3-L-S305A)] were separated on reducing 12% SDS-PAGE gels and analysed using western blotting with HtrA3 mAbs (6G6, 9C9, 2C4, 10H10 and 3E6), anti-HtrA1 and anti-HtrA2 antibodies respectively. ( B ) Western blot analysis of neat conditioned media from HEK-293F cells transfected with control plasmids (control transfection, C) or WT HtrA3-L (L) or HtrA3-S (S) constructs with the five HtrA3 mAbs. The cleaved forms of HtrA3 are indicated as HtrA3-L/C and HtrA3-S/C.

Data is shown for three independent AlphaLISAs employing different mAb pairs [denoted as conjugated (C)/biotinylated (B)]: 10H10C/6G6B, 10H10C/2C4B and 10H10C/3E6B . Assays were optimised using HtrA3-L-S305A as the standard with the 1 h (A–C) and overnight (D and E) protocols. ( A ) Detection of an equal amount (50 pM) of HtrA3, HtrA2, HtrA1 or BSA, or measuring HtrA3 (50 pM) when either a biotinylated irrelevant antibody (unrelated mAb) replaced the biotinylated HtrA3 mAb or donor beads were absent. ( B ) Analysis of an equal amount of HtrA3 (50 pM) with various concentrations of biotinylated mAbs to determine the sub-hook point. ( C ) Serum matrix inhibition of the 10H10C/2C4B AlphaLISA. Serum was titrated with an equal amount (500 pM) of HtrA3. ( D ) Representative calibration curves of HtrA3-L-S305A in IB and serum using the overnight protocol. ( E ) Analysis of WT HtrA3 proteins with the overnight AlphaLISA protocol. Conditioned media from HEK-293F cells transfected with WT HtrA3-L or HtrA3-S constructs were assayed, HtrA3 concentrations were determined from a calibration curve of HtrA3-L-S305A standard diluted in transfection medium.