Anti-PPCEL/PREPL antibody [EPR17751] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PPCEL/PREPL antibody [EPR17751] - BSA and Azide free (AB251361)

This data was developed using ab202064, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized SH-SY5Y (Human neuroblastoma from bone marrow cells) cells labeling PPCEL/PREPL with ab202064 at 1/1000, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/500 (green). Confocal image showing cytoplasmic staining on SH-SY5Y cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 and Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) at 1/500 (red).
The negative controls are as follows : -
-ve control 1 - ab202064 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/500 dilution.
-ve control 2. - Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/500 dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-PPCEL/PREPL antibody [EPR17751] - BSA and Azide free (AB251361)

This data was developed using ab202064, the same antibody clone in a different buffer formulation.

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NIH/3T3 (Mouse embryo fibroblast) cells labeling PPCEL/PREPL with ab202064 at 1/150 (red) compared with a rabbit monoclonal IgG isotype control (ab172730) (black) and a unlabelled control (cells without incubation with primary antibody and secondary antibody (blue)). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

• WB

Supplier Data

Western blot - Anti-PPCEL/PREPL antibody [EPR17751] - BSA and Azide free (AB251361)

This data was developed using ab202064, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PPCEL/PREPL antibody [EPR17751] (<a href='/en-us/products/primary-antibodies/ppcel-prepl-antibody-epr17751-ab202064'>ab202064</a>) at 1/5000 dilution

All lanes:

Mouse brain lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 84 kDa

Observed band size: 72 kDa

true

Exposure time: 5s

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-PPCEL/PREPL antibody [EPR17751] - BSA and Azide free (AB251361)

This data was developed using ab202064, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 100% methanol-fixed, 0.1% Triton X-100 permeabilized MCF7 (Human breast adenocarcinoma cell line) cells labeling PPCEL/PREPL with ab202064 at 1/1000, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 (green). Confocal image showing cytoplasmic staining on MCF-7 cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary antibody at 1/500 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab202064 at 1/1000 dilution, followed by Goat Anti-Mouse IgG H&L (Alexa Fluor^® 594) (ab150120) secondary at 1/500 dilution.
-ve control 2 : Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary at 1/500 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-PPCEL/PREPL antibody [EPR17751] - BSA and Azide free (AB251361)

This data was developed using ab202064&44; the same antibody clone in a different buffer formulation.

PPCEL/PREPL was immunoprecipitated from 1mg of MCF7 (Human breast adenocarcinoma cell line) whole cell extract with ab202064 at 1/40.

Western blot was performed using ab202064 at 1/2000.

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1 : MCF7 whole cell lysate at 10ug.
Lane 2 : Immunoprecipitate of MCF7 whole cell lysate using ab202064.
Lane 3 : Immunoprecipitate of MCF7 whole cell lysate using ab172730 (Rabbit monoclonal IgG).
Blocking and dilution buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-PPCEL/PREPL antibody [EPR17751] (<a href='/en-us/products/primary-antibodies/ppcel-prepl-antibody-epr17751-ab202064'>ab202064</a>)

Predicted band size: 84 kDa

Observed band size: 72 kDa

true

Exposure time: 8s

• WB

Supplier Data

Western blot - Anti-PPCEL/PREPL antibody [EPR17751] - BSA and Azide free (AB251361)

This data was developed using ab202064, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PPCEL/PREPL antibody [EPR17751] (<a href='/en-us/products/primary-antibodies/ppcel-prepl-antibody-epr17751-ab202064'>ab202064</a>) at 1/5000 dilution

Lane 1:

A673 (Human muscle Ewing's Sarcoma cell line) lysate at 20 µg

Lane 2:

MCF7 (Human breast adenocarcinoma cell line ) lysate at 20 µg

Lane 3:

L6 (Rat skeletal muscle cell line) lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 84 kDa

Observed band size: 72 kDa

true

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-PPCEL/PREPL antibody [EPR17751] - BSA and Azide free (AB251361)

This data was developed using ab202064, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PPCEL/PREPL antibody [EPR17751] (<a href='/en-us/products/primary-antibodies/ppcel-prepl-antibody-epr17751-ab202064'>ab202064</a>) at 1/1000 dilution

Lane 1:

C6 (Rat glial tumor cells) lysate at 10 µg

Lane 2:

RAW264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) lysate at 10 µg

Lane 3:

PC-12 (Rat adrenal gland pheochromocytoma) lysate at 10 µg

Lane 4:

NIH/3T3 (Mouse embryo fibroblast cells) lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 84 kDa

Observed band size: 72 kDa

true

Exposure time: 5s

• WB

Supplier Data

Western blot - Anti-PPCEL/PREPL antibody [EPR17751] - BSA and Azide free (AB251361)

This data was developed using ab202064, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PPCEL/PREPL antibody [EPR17751] (<a href='/en-us/products/primary-antibodies/ppcel-prepl-antibody-epr17751-ab202064'>ab202064</a>) at 1/1000 dilution

Lane 1:

Human fetal brain tissue lysate at 10 µg

Lane 2:

Human fetal heart tissue lysate at 10 µg

Lane 3:

Human fetal kidney tissue lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 84 kDa

Observed band size: 72 kDa

true

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-PPCEL/PREPL antibody [EPR17751] - BSA and Azide free (AB251361)

This data was developed using ab202064, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-PPCEL/PREPL antibody [EPR17751] (<a href='/en-us/products/primary-antibodies/ppcel-prepl-antibody-epr17751-ab202064'>ab202064</a>) at 1/10000 dilution

All lanes:

Recombinant Human PPCEL/PREPL protein (<a href='/en-us/products/unavailable/recombinant-human-ppcelprepl-protein-ab160557'>ab160557</a>)

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: 84 kDa

Observed band size: 110 kDa

true

Exposure time: 3s