Immunofluorescent analysis of Alpha-1-Antitrypsin was performed using 70% confluent log phase Hep G2 cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Alpha-1-Antitrypsin Rabbit Polyclonal Antibody (Product # PA5-16661) at 1:250 dilution in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Rabbit IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conjugate (Product # A27034) a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Horse, Human, Mink, Non-human primate|
|Published species reactivity||Mouse|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Purified human serum alpha-1-AT|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||1:20|
|Immunohistochemistry (Paraffin) (IHC (P))||1:100-1:200|
|Western Blot (WB)||1:250|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Immunohistochemistry (IHC)||See 1 publications below|
PA5-16661 targets Alpha-1-Antitrypsin in IHC (P) applications and shows reactivity with Equine, Human, Mink, monkey, and baboon samples. Does not react with Canine, Chicken, Feline, Guinea Pig, Goat, Marsupial, mouse, Ovine, and Porcine samples
The PA5-16661 immunogen is purified human serum alpha-1-AT.
alpha-1-antitrypsin (alpha-1-AT) which exists in a number of genetic variants. MM variant is the most common. Alpha-1-AT is synthesized in the liver and it acts as an inhibitor of proteases such as trypsin, elastase, chymotrypsin, collagenase, leucocytic proteases, plasmin, and thrombin, which may be released during inflammatory reactions in the lung. In the absence of alpha-1-AT, these enzymes are not inhibited and they may digest pulmonary parenchyma. Alpha-1-AT deficiency is associated with chronic obstructive lung disease (emphysema) and less frequently with hepatic cirrhosis in infants and respiratory distress of the newborn. Increase in alpha-1-AT occurs as an acute phase response to tissue necrosis and inflammation. Serum level of alpha-1-AT is elevated in rheumatoid arthritis, bacterial infections, vasculitis, and carcinomatosis.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Isolation of adult human pluripotent stem cells from mesenchymal cell populations and their application to liver damages.
PA5-16661 was used in immunohistochemistry to study the isolation of adult human pluripotent stem cells from mesenchymal cell populations and their application in liver damage
|Wakao S,Kitada M,Kuroda Y,Dezawa M||Methods in molecular biology (Clifton, N.J.) (826:89)||2011|