The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/1000. Predicted molecular weight: 47 kDa.
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Although its physiological function is unclear, it can inhibit neutrophil cathepsin G and mast cell chymase, both of which can convert angiotensin-1 to the active angiotensin-2.
Plasma. Synthesized in the liver. Like the related alpha-1-antitrypsin, its concentration increases in the acute phase of inflammation or infection. Found in the amyloid plaques from the hippocampus of Alzheimer disease brains.
Involvement in disease
Defects in SERPINA3 may be a cause of chronic obstructive pulmonary disease (COPD) [MIM:107280].
Belongs to the serpin family.
The reactive center loop (RCL) extends out from the body of the protein and directs binding to the target protease. The protease cleaves the serpin at the reactive site within the RCL, establishing a covalent linkage between the carboxyl group of the serpin reactive site and the serine hydroxyl of the protease. The resulting inactive serpin-protease complex is highly stable.
Alpha 1 Antichymotrypsin was immunoprecipitated from 1 mg of human blood lysate with ab204514 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab204514 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/1500 dilution.
Lane 1: Human blood lysate 10 µg (Input).
Lane 2: ab204514 IP in human blood lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab204514 in human blood lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 30 seconds.
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Raji (human Burkitt's lymphoma cell line) cells labeling alpha 1 Antichymotrypsin with ab204514 at 1/200 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic and nuclear staining on Raji cell line.
The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) at 1/1000 dilution (red).
The negative controls are as follows: -ve control 1: ab204214 at 1/200 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary at 1/1000 dilution. -ve control 2: Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary at 1/1000 dilution.
Immunohistochemical analysis of paraffin-embedded human tonsil tissue labeling alpha 1 Antichymotrypsin with ab204514 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic staining on human tonsil tissue is observed. Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) ab97051 at 1/500 dilution.
Western blot - Anti-alpha 1 Antichymotrypsin antibody [EPR17088] (ab204514)
All lanes : Anti-alpha 1 Antichymotrypsin antibody [EPR17088] (ab204514) at 1/1000 dilution
Lane 1 : Human plasma Lane 2 : Human plasma deglycosylation (PNGase F) treated
Lysates/proteins at 10 µg per lane.
Secondary Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution