Western blot analysis was performed on membrane enriched extracts (30 ug) of (Fig 1) HT-29 (Lane 1) and Hep G2 (Lane 2).Likewise, Western blot analysis was performed on (Fig 2) condition media of Hep G2 (Lane 1), PC3 (Lane 2) and HeLa (Lane 3).The blots were probed with Anti-Alpha1-Antitrypsin Mouse Monoclonal Antibody (Product # MA5-14661, 2 ug/mL) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.4 ug/ml, 1:2500 dilution). In Figure 1, a ~47 kDa band corresponding Alpha1-Antitrypsin was observed in Hep G2, and a ~55kDa band was observed in HT-29 due to glycosylation of protein. Also, in Figure 2, ~60kDa bands were observed in Hep G2, PC 3 and HeLa due to glycosylation of protein. Known quantity of protein samples were electrophoresed using Novex® NuPAGE®4-12 % Bis-Tris gel (Product # NP0321BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane by iBlot® 2 Dry Blotting System (Product # IB21001). The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product # SLF2000S). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
|Tested species reactivity||Human, Mouse|
|Host / Isotype||Mouse / IgG2a|
|Immunogen||Purified human a1-Antitrypsin|
|Storage buffer||0.01M potassium phosphate with 2.5mM KCl, 0.135M NaCl, 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 6 months. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|ELISA (ELISA)||Assay Dependent|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunoprecipitation (IP)||Assay Dependent|
|Radioimmune Assays (RIA)||Assay Dependent|
|Western Blot (WB)||1-3 µg/ml|
* 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.
MA5-14661 targets Alpha1-Antitrypsin in ELISA, IP, and RIA applications and shows reactivity with Human samples.
The MA5-14661 immunogen is purified human a1-Antitrypsin.
MA5-14661 detects Alpha1-Antitrypsin which has a predicted molecular weight of approximately 46 kDa.
Product MA514661 is a smaller package size of MIA1101 (formerly sold as a Seradyn product).
The protein encoded by this gene is secreted and is a serine protease inhibitor whose targets include elastase, plasmin, thrombin, trypsin, chymotrypsin, and plasminogen activator. Defects in this gene can cause emphysema or liver disease. Several transcript variants encoding the same protein have been found for this gene.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.