- MA5-11945 - Invitrogen Antibodies PRTN3 antibody

Submitted references 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) 2012 May;35(4):309-20

A hydrophobic patch on proteinase 3, the target of autoantibodies in Wegener granulomatosis, mediates membrane binding via NB1 receptors.

Korkmaz B, Kuhl A, Bayat B, Santoso S, Jenne DE
The Journal of biological chemistry 2008 Dec 19;283(51):35976-82

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western blot analysis of Proteinase 3 was performed by loading 25 µg 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 4ºC 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 ~28 kDa.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Proteinase 3 in BAF-3 cells (green) compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/mL, fixed with 2% paraformaldehyde and washed with PBS. Cells were blocked with a 2% solution of BSA-PBS for 30 min at room temperature and incubated with a Proteinase 3 monoclonal antibody (Product # MA5-11945) at a dilution of 1 µg/test for 40 min at room temperature. Cells were then incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated secondary antibody and re-suspended in PBS for FACS analysis.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Proteinase 3 showing staining in PBMC cells (green) compared to an isotype control (blue). Human blood was collected, combined with a hydrophilic polysaccharide, centrifuged, transferred to a conical tube and washed with PBS. 50 µL of cell solution was added to each tube at a dilution of 2x10^7 cells/mL, followed by the addition of 50 µL of isotype control and primary antibody (Product # MA5-11945) at a dilution of 1 µg/test. Cells were incubated for 30 min at 4ºC and washed with a cell buffer, followed by incubation with a DyLight 488-conjugated goat anti-mouse IgG (H+L) secondary for 30 min at 4ºC in the dark. FACS analysis was performed using 400 µL of cell buffer.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Flow cytometry analysis of Proteinase 3 showing staining in PBMC cells (green) compared to an isotype control (blue). Human blood was collected, combined with a hydrophilic polysaccharide, centrifuged, transferred to a conical tube and washed with PBS. 50 µL of cell solution was added to each tube at a dilution of 2x10^7 cells/mL, followed by the addition of 50 µL of isotype control and primary antibody (Product # MA5-11945) at a dilution of 1 µg/test. Cells were incubated for 30 min at 4ºC and washed with a cell buffer, followed by incubation with a DyLight 488-conjugated goat anti-mouse IgG (H+L) secondary for 30 min at 4ºC in the dark. FACS analysis was performed using 400 µL of cell buffer.

MA5-11945

Antibody data