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Active Alpha-1-Antitrypsin (a1AT)

SERPINA1; SPAAT; A1-AT; PI; Serpin Peptidase Inhibitor,Clade A(Alpha-1 Antiproteinase/AntiTrypsin)Member 1; Alpha-1 protease inhibitor; Short peptide from AAT; Serpin A1

ACTIVITY TEST

Alpha-1-Antitrypsin (a1AT), a major serine protease inhibitor synthesized primarily in the liver, is crucial for protecting tissues from excessive proteolytic damage. Circulating in human plasma, it targets and inhibits a range of proteases—including neutrophil elastase, trypsin, and chymotrypsin—which, if left unregulated, can degrade essential structural proteins. Beyond its protective function, A1AT is involved in modulating inflammatory responses and maintaining pulmonary homeostasis; deficiencies in A1AT are associated with conditions such as emphysema and liver disorders. Notably, A1AT can directly bind to and inhibit Renin (REN), a central enzyme in the renin-angiotensin system, thereby modulating its proteolytic activity.To detect the activity of recombinant a1AT , a functional ELISA assay was performed to evaluate the interaction between recombinant mouse a1AT and recombinant dog REN.Briefly, a1AT was diluted serially in PBS with 0.01% BSA (pH 7.4). Duplicate samples of 100 μl were then transferred to REN-coated microtiter wells and incubated for 1h at 37℃. Wells were washed with PBST and incubated for 1h with anti-a1AT pAb, then aspirated and washed 3 times. After incubation with HRP labelled secondary antibody for 1h at 37℃, wells were aspirated and washed 5 times. With the addition of substrate solution, wells were incubated 15-25 minutes at 37℃. Finally, add 50 µL stop solution to the wells and read at 450/630nm immediately. The binding activity of recombinant mouse a1AT and recombinant dog REN was shown in Figure 1, the EC50 for this effect is 0.054 µg/mL..

USAGE

Reconstitute in 10mM PBS (pH7.4) to a concentration of 0.1-1.0 mg/mL. Do not vortex.

STORAGE

Avoid repeated freeze/thaw cycles. Store at 2-8°C for one month. Aliquot and store at -80°C for 12 months.

STABILITY

The thermal stability is described by the loss rate. The loss rate was determined by accelerated thermal degradation test, that is, incubate the protein at 37°C for 48h, and no obvious degradation and precipitation were observed. The loss rate is less than 5% within the expiration date under appropriate storage condition.

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Magazine Citations
Journal of Biotechnology Expression and characterization of recombinant human alpha-antitrypsin in transgenic rice seed PubMed: 23376844
Sci Rep Hepatic steatosis depresses alpha-1-antitrypsin levels in human and rat acute pancreatitis PubMed: 26634430
PLoS One Gut Microbial Dysbiosis May Predict Diarrhea and Fatigue in Patients Undergoing Pelvic Cancer Radiotherapy: A Pilot Study PubMed: 25955845
Respir Care Diagnostic Utility of Biomarkers in COPD PubMed: 26106205
Acta Anaesthesiologica Scandinavica Early prognostic factors in septic shock cancer patients: a prospective study with a proteomic approach Pubmed:29315472
Journal of Clinical Medicine Urinary Proteomics for the Early Diagnosis of Diabetic Nephropathy in Taiwanese Patients Pubmed: 30486327
Journal of Proteomics Salivary proteins from dysplastic leukoplakia and oral squamous cell carcinoma and their potential for early detection Pubmed: 31706945
Diagnostics Systemic Alterations of Immune Response-Related Proteins during Glaucoma Development in the Murine Model DBA/2J Pubmed: 32585848
Pediatrics International Prenatal diagnosis of maternal serum from mothers carrying β‐thalassemic fetus 34559910
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