Western blot analysis of Proteinase 3 was performed by loading 25 ug of HL60 (lane 1), human spleen (lane 2) and mouse brain (lane 3) lysates onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4°C overnight. The membrane was probed with a Proteinase 3 monoclonal antibody (Product # MA5-11945) at a dilution of 1:100 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~28kDa.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Human|
|Host / Isotype||Mouse / IgG1, kappa|
|Immunogen||HMC-1/PR3 cell granule extract|
|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)||1ug/test|
|Immunofluorescence (IF)||Assay Dependent|
|Western Blot (WB)||1:50-1:500|
* 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.
MA5-11945 targets Proteinase 3 in IF, FACS and WB applications and shows reactivity with mouse and human samples.
The MA5-11945 immunogen is hMC-1/PR3 cell granule extract.
Proteinase 3 (PR3), a constituent of azurophil granules of neutrophils (polymorphonuclear cells, PMNs), is the target antigen for most anti-neutrophil cytoplasmic antibodies (c-ANCA) in Wegener's granulomatosis (WG).
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
The role of antigen cross-presentation from leukemia blasts on immunity to the leukemia-associated antigen PR1.
MA5-11945 was used in ELISA, immunocytochemistry, and western blot to study the role of antigen cross-presentation from leukemia blasts on immunity to the leukemia-associated antigen PR1
|Alatrash G,Ono Y,Sergeeva A,Sukhumalchandra P,Zhang M,St John LS,Yang TH,Ruisaard K,Armistead PM,Mittendorf EA,He H,Qiao N,Rodriguez-Cruz T,Liang S,Clise-Dwyer K,Wieder ED,Lizee G,Lu S,Molldrem JJ||Journal of immunotherapy (Hagerstown, Md. : 1997) (35:309)||2012|
A hydrophobic patch on proteinase 3, the target of autoantibodies in Wegener granulomatosis, mediates membrane binding via NB1 receptors.
MA5-11945 was used in flow cytometry to investigate the role of a specific roteinase 3 domain on membrane binding and its mechanism
|Korkmaz B,Kuhl A,Bayat B,Santoso S,Jenne DE||The Journal of biological chemistry (283:35976)||2008|