Anti-Cathepsin D [EPR3057Y] - Mouse IgG2a (Chimeric)

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-Cathepsin D [EPR3057Y] - Mouse IgG2a (Chimeric) (AB302649)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling CTSD (cathepsin D) with ab302649 at 1/1000 dilution (0.1ug) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Cathepsin D [EPR3057Y] - Mouse IgG2a (Chimeric) (AB302649)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling CTSD (cathepsin D) with ab302649 at 1/50 (18.84 ug/ml) dilution, followed by ab98736 Goat F(ab')2 Anti-Mouse IgG - Fc (DyLight® 488) pre-adsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic staining in HepG2 cell line is observed. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 10ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab98736 Goat F(ab')2 Anti-Mouse IgG - Fc (DyLight® 488) pre-adsorbed at 1/1000 2 ug/ml dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin D [EPR3057Y] - Mouse IgG2a (Chimeric) (AB302649)

Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling CTSD (cathepsin D) with ab302649 at 1/10000 (0.094 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic with granular staining on human breast. The section was incubated with ab302649 for 30 mins at room temperature, followed by anti-mouse IgG2a antibody for 8 mins during the LeicaDS9800 kit staining procedure.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cathepsin D [EPR3057Y] - Mouse IgG2a (Chimeric) (AB302649)

Immunohistochemical analysis of paraffin-embedded Human liver tissue labeling CTSD (cathepsin D) with ab302649 at 1/10000 (0.094 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic with granular staining on human liver. The section was incubated with ab302649 for 30 mins at room temperature, followed by anti-mouse IgG2a antibody for 8 mins during the LeicaDS9800 kit staining procedure.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Cathepsin D [EPR3057Y] - Mouse IgG2a (Chimeric) (AB302649)

Immunofluorescent analysis of 100% methanol-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling CTSD (cathepsin D) with ab302649 at 1/50 (18.84 ug/ml) dilution, followed by ab98736 Goat F(ab')2 Anti-Mouse IgG - Fc (DyLight® 488) pre-adsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing cytoplasmic staining in Neuro-2a cell line is observed. ab179513 Anti-beta Tubulin rabbit monoclonal antibody was used to counterstain tubulin at 1/200 10ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab98736 Goat F(ab')2 Anti-Mouse IgG - Fc (DyLight® 488) pre-adsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Cathepsin D [EPR3057Y] - Mouse IgG2a (Chimeric) (AB302649)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling CTSD (cathepsin D) with ab302649 at 1/1000 dilution (0.1ug) (Red) compared with a Mouse monoclonal IgG (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti-Mouse IgG (Alexa Fluor® 488, ab150113) at 1/2000 dilution was used as the secondary antibody.

• WB

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Western blot - Anti-Cathepsin D [EPR3057Y] - Mouse IgG2a (Chimeric) (AB302649)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : Lane 1-3 : 15 seconds Lane 4, 5 : 125 seconds

All lanes:

Western blot - Anti-Cathepsin D [EPR3057Y] - Mouse IgG2a (Chimeric) (ab302649) at 1/1000 dilution

Lane 1:

Human heart tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human spleen tissue lysate at 20 µg

Lane 4:

Mouse brain tissue lysate at 20 µg

Lane 5:

Rat stomach tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG Fc (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-fc-hrp-preadsorbed-ab98717'>ab98717</a>) at 1/20000 dilution

Observed band size: 14 kDa,28 kDa,46 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Cathepsin D [EPR3057Y] - Mouse IgG2a (Chimeric) (AB302649)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Exposure time : 26 seconds

All lanes:

Western blot - Anti-Cathepsin D [EPR3057Y] - Mouse IgG2a (Chimeric) (ab302649) at 1/1000 dilution

Lane 1:

MCF7 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate at 20 µg

Lane 3:

SK-BR-3 (human breast adenocarcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

U937 (human histiocytic lymphoma monocyte), whole cell lysate at 20 µg

Lane 5:

A431 (human epidermoid carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 6:

HepG2 (human hepatocellular carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 7:

RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 20 µg

Lane 8:

Neuro-2a (mouse neuroblastoma neuroblast), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG Fc (HRP) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-mouse-igg-fc-hrp-preadsorbed-ab98717'>ab98717</a>) at 1/20000 dilution

Observed band size: 28 kDa,14 kDa,46 kDa

false

Exposure time: 26s