Anti-Proteinase 3/Myeloblastin antibody [EPR29801-588]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Proteinase 3/Myeloblastin antibody [EPR29801-588] (AB324528)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized K-562 (human chronic myelogenous leukemia lymphoblast, Left) / HL-60 (human acute promyelocytic leukemia promyeloblast, Right) cells labelling Proteinase 3/Myeloblastin with ab324528 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Negative control : K-562 (PMID : 10572086).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Proteinase 3/Myeloblastin antibody [EPR29801-588] (AB324528)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized WEHI-3 (mouse leukemia lymphoblast) cells labelling Proteinase 3/Myeloblastin with ab324528 at 1/200 (2.445 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in WEHI-3 cell line (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Low expression : EL4.

ab7291 Anti-alpha Tubulin mouse monoclonal antibody was used to counterstain tubulin at 1/1000 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab324528 at 1/200 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2 : ab7291 at 1/1000 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Proteinase 3/Myeloblastin antibody [EPR29801-588] (AB324528)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized EL4 (mouse lymphoma T lymphocyte, Left) / WEHI-3 (mouse leukemia lymphoblast, Right) cells labelling Proteinase 3/Myeloblastin with ab324528 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

Low expression : EL4.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Proteinase 3/Myeloblastin antibody [EPR29801-588] (AB324528)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Mouse bone marrow cells labelling Proteinase 3/Myeloblastin with ab324528 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody).

Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Proteinase 3/Myeloblastin antibody [EPR29801-588] (AB324528)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Mouse bone marrow cells labelling Proteinase 3/Myeloblastin with ab324528 at 1/200 (2.445 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing cytoplasmic staining in CD11b+ cells of mouse bone marrow (shown in green). The counterstain was observed in magenta. Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

ab8878 Anti-CD11b Rat monoclonal antibody was used to counterstain tubulin at 1/200 dilution (Magenta). The Nuclear counterstain was DAPI (Blue).

-ve control 1 : ab324528 at 1/200 dilution, followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) antibody at 1/1000 dilution.
-ve control 2 : ab8878 at 1/200 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution.

• I-ELISA

Supplier Data

Indirect ELISA - Anti-Proteinase 3/Myeloblastin antibody [EPR29801-588] (AB324528)

Indirect ELISA analysis of ab324528 at 1000-0 ng/ml. The Secondary antibody used was Alkaline Phosphatase-conjugated AffiniPure Goat Anti-Rabbit IgG (H+L) at 1 : 2500 dilution dilution.

Antigen : Mouse PR3 fragment, Mouse Elastase-2 fragment.

Antigen concentration : 1000 ng/ml

This antibody does not cross-react with Mouse Elastase-2.