Anti-Myeloperoxidase antibody [EPR17996]

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Myeloperoxidase antibody [EPR17996] (AB188211)

Flow cytometry staining of C57 BL/6 mouse bone marrow cells with ab188211 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were fixed and permeabilised with BD Cytofix/Cytoperm™ for 20 min. Cells were incubated for 30min at 22°C in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab188211 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100 μl at 0.2 μg/ml (1/10200)) for 30min at 22°C. The cells were simultaneously stained with Ly6G.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myeloperoxidase antibody [EPR17996] (AB188211)

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling Myeloperoxidase with ab188211 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on neutrophils of human spleen [PMID : 19566938].

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Myeloperoxidase antibody [EPR17996] (AB188211)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed mouse PBMC labeling Myeloperoxidase with ab188211 at 1/500 dilution (Right) compared with a rabbit monoclonal IgG isotype control (ab172730; Left). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.

Mouse peripheral blood mononuclear cells stained intracellularly with ab188211 (Right) and isotype control (Left). Only monocytes and granulocytes (larger SSC population) result in positive signal while the lymphocyte population remains unchanged.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myeloperoxidase antibody [EPR17996] (AB188211)

Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling Myeloperoxidase with ab188211 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on neutrophils of rat spleen [PMID : 19566938].

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myeloperoxidase antibody [EPR17996] (AB188211)

Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling Myeloperoxidase with ab188211 at 1/8000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use. Cytoplasmic staining on neutrophils of mouse spleen [PMID : 19566938].

Counter stained with Hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-Myeloperoxidase antibody [EPR17996] (AB188211)

Blocking/Dilution buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature PMID : 2154223.

All lanes:

Western blot - Anti-Myeloperoxidase antibody [EPR17996] (ab188211) at 1/1000 dilution

All lanes:

Rat spleen lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 83 kDa

Observed band size: 13 kDa,74 kDa

false

Exposure time: 30s

• WB

Supplier Data

Western blot - Anti-Myeloperoxidase antibody [EPR17996] (AB188211)

Blocking/Dilution buffer : 5% NFDM/TBST.

The molecular weight observed is consistent with what has been described in the literature. PMID : 2154223. 89 kDa (MPO), 74 kDa (intermediate form), 13 kDa (light chain)

Negative control : NIH/3T3 PMID : 9001423.

All lanes:

Western blot - Anti-Myeloperoxidase antibody [EPR17996] (ab188211) at 1/1000 dilution

Lane 1:

Mouse spleen lysate at 20 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 83 kDa

Observed band size: 13 kDa,74 kDa,89 kDa

false

Exposure time: 30s