Anti-proCathepsin D antibody [EPR24352-13] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-proCathepsin D antibody [EPR24352-13] - BSA and Azide free (AB282025)

This data was developed using ab270999, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Neuro-2a (Mouse neuroblastoma neuroblast) cells labelling pro-Cathepsin D with ab270999 at 1/50 dilution (1μg) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-proCathepsin D antibody [EPR24352-13] - BSA and Azide free (AB282025)

This data was developed using ab270999, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized Neuro-2a cells labelling pro-Cathepsin D with ab270999 at 1/50 (11.48 μg/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in Neuro-2a cells is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-proCathepsin D antibody [EPR24352-13] - BSA and Azide free (AB282025)

This data was developed using ab270999, the same antibody clone in a different buffer formulation.

pro-Cathepsin D was immunoprecipitated from 0.35 mg RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 μg with ab270999 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270999 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : RAW 264.7 whole cell lysate 10 μg

Lane 2 : ab270999 IP in RAW 264.7 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab270999 in RAW 264.7 whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 67 seconds

All lanes:

Immunoprecipitation - Anti-proCathepsin D antibody [EPR24352-13] (<a href='/en-us/products/primary-antibodies/procathepsin-d-antibody-epr24352-13-ab270999'>ab270999</a>)

Predicted band size: 45 kDa

Observed band size: 48 kDa

false

• WB

Lab

Western blot - Anti-proCathepsin D antibody [EPR24352-13] - BSA and Azide free (AB282025)

This data was developed using ab270999, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration : 5% NFDM/TBST

The molecular weight observed is consistent with what has been described in the literature (PMID : 28453791).

Exposure time : 3 minutes

All lanes:

Western blot - Anti-proCathepsin D antibody [EPR24352-13] (<a href='/en-us/products/primary-antibodies/procathepsin-d-antibody-epr24352-13-ab270999'>ab270999</a>) at 1/1000 dilution

Lane 1:

RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 2:

Neuro-2a (mouse neuroblastoma neuroblast) 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: 45 kDa

Observed band size: 48 kDa

false