Description: Alpha-1-antitrypsin is synthesised in the liver and is present in serum and tissue fluids where it acts as an inhibitor of proteases, particularly elastase. Its main function appears to be the neutralisation of elastase released by neutrophils during an inflammatory response. As a result, in alpha-1-antitypsin deficiency, any stimulus which activates neutrophils in the lung may result in uninhibited elastase-induced tissue destruction.
Primary Antibody
Name: alpha-1-Antitrypsin Antibody |
Clone: Rabbit Polyclonal |
Supplier: Novocastra Labs |
Catalog Number: NCL-A1Ap |
Dilution: 1:250 - 1:500 using IHC-TekTM Antibody Diluent (Cat# IW-1000 or IW-1001) to reduce background and unspecific staining and serum blocking step is NOT needed. |
Incubation Time/Temp: 60 min/room temperature |
Device: Incubator |
Buffer/pH value: IHC-TekTM Proteinase K Solution (Cat# IW-1101) |
Heat/Cool Temperature: 37 ºC/room temperature |
Heat/Cool Time: 20 minutes/20 minutes |
Standard Method: ABC Method or LSAB Method |
Enhanced Method: Polymeric Methods |
Reagent: DAB |
Incubation Time/Temperature: 1-3 minutes/room temperature |
Reagent: Mayer's Hematoxylin |
Staining Time: 30 seconds |
Staining Pattern: Cytoplasmic staining of macrophages |
Images: Search image |
Tissue Type: Colon, tonsil |
Fixation: Formalin-fixed paraffin sections, and acetone fixed frozen sections |
Positive Control: Colon, tonsil |
Negative Control: Omit primary antibody, isotype control, absorption control |
Blocking: 2-5% normal serum to reduce unspecific background staining; 0.5-3% H2O2 to block endogenous peroxidase activity; avidin/biotin to block endogenous biotin activity if necessary |
There is no need to perform antigen retrieval on acetone fixed frozen sections |
References:
1. Alpha-1-antitrypsin deficiency - Alpha-1 antitrypsin deficiency (A1AD) is an autosomal recessive disorder marked by abnormally low or no serum levels of alpha-1 antitrypsin in the blood. It is the most common genetic cause of liver disease in children and of emphysema in adults
2. Callea F, Fevery J, De Groote J, Desmet VJ. Detection of Pi Z phenotype individuals by alpha-1-antitrypsin immunohistochemistry in paraffin-embedded liver tissue specimens. J Hepatol 1986; 2:389-401. PubMed Abstract.
3. Carlson J, Eriksson S. Chronic 'cryptogenic' liver disease and malignant hepatoma in intermediate alpha-1-antitrypsin deficiency identified by a Pi Z-specific monoclonal antibody. Scand J Gastroenterol. 1985; 20:835-842. PubMed Abstract.