Anti-Myeloperoxidase antibody

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Myeloperoxidase antibody (AB45977)

ab45977 stained in HL60 cells. Cells were fixed with 100% methanol (5min) at room temperature and incubated with PBS containing 10% goat serum, 0.3 M glycine, 1% BSA and 0.1% tween for 1h at room temperature to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab45977 at 5μg/ml and ab72791 (Mouse monoclonal to alpha Tubulin - Loading Control) at 1ug/ml overnight at +4°C. The secondary antibodies used were ab150081 used at 1 ug/ml (colored green) and ab150116 (pseudo-colored red) used at 2ug/ml for 1hour at room temperature. DAPI was used to stain the cell nuclei (colored blue) at a concentration of 1.43μM for 1hour at room temperature.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myeloperoxidase antibody (AB45977)

IHC image of 45977 staining in human tonsil formalin fixed paraffin embedded tissue section, performed on a Leica Bond^TM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab45977, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

• IHC-P

AbReview11726****

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Myeloperoxidase antibody (AB45977)

ab45977 (1/50) staining Myeloperoxidase in paraffin-embbeded human spleen tissue sections. Tissue underwent fixation in formaldehyde, heat-mediated antigen retrieval in citrate buffer pH 6.0 and blocking (5 minutes/peroxidase block and 10 minutes/protein block). For further experimental details please refer to abreview.

This image is courtesy of an Abreview submitted by Antibody Solutions Ltd.

• WB

Lab

Western blot - Anti-Myeloperoxidase antibody (AB45977)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab45977 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406

All lanes:

Western blot - Anti-Myeloperoxidase antibody (ab45977) at 1 µg/mL

All lanes:

HL60 (Human promyelocytic leukemia 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 1/50000 dilution

Predicted band size: 83 kDa

Observed band size: 84 kDa

true

Exposure time: 2min

• WB

Unknown

Western blot - Anti-Myeloperoxidase antibody (AB45977)

The mature myeloperoxidase protein is a tetramer of two heavy chains (60 kDa) and two light chains (12 kDa). Our immunogen sequence is within the myeloperoxidase light chain. In HL60 cells, ab45977 detects bands at approximately 84-kDa, corresponding to the expected MW of full-length Myeloperoxidase protein, and at 12-kDa, corresponding to the expected MW of myeloperoxidase light chain.

All lanes:

Western blot - Anti-Myeloperoxidase antibody (ab45977) at 1 µg/mL

All lanes:

HL60 (Human promyelocytic leukemia cell line) Whole Cell Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-preadsorbed-ab97080'>ab97080</a>) at 1/5000 dilution

Predicted band size: 83 kDa

Observed band size: 12 kDa,35 kDa,75 kDa,84 kDa

false

Exposure time: 3min