Anti-PPCEL/PREPL antibody [EPR17752]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPCEL/PREPL antibody [EPR17752] (AB203111)

Immunohistochemical analysis of paraffin-embedded Human cerebral cortex tissue labeling PPCEL/PREPL with ab203111 at 1/250 dilution followed by ab97051 Goat Anti-Rabbit IgG H&L (HRP) at a 1/500 dilution. Cytoplasmic staining on Human cerebral cortex tissue is observed. Counter stained with Hematoxylin.

Negative control : Used PBS instead of primary antibody secondary antibody is ab97051 Goat Anti-Rabbit IgG H&L (HRP).

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

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PPCEL/PREPL antibody [EPR17752] (AB203111)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized MCF-7(Human breast adenocarcinoma cell line) cells labeling PPCEL/PREPL with ab203111 at 1/250 dilution followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on MCF-7 cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows : -
-ve control 1 - ab203111 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-PPCEL/PREPL antibody [EPR17752] (AB203111)

Immunofluorescent analysis of 4% paraformaldehyde-fixed 0.1% Triton X-100 permeabilized SH-SY5Y (Human neuroblastoma from bone marrow cells) cells labeling PPCEL/PREPL with ab203111 at 1/250 dilution followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on SH-SY5Y cell line. The nuclear counterstain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

The negative controls are as follows : -
-ve control 1 - ab203111 at 1/250 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
-ve control 2. - ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PPCEL/PREPL antibody [EPR17752] (AB203111)

Immunohistochemical analysis of paraffin-embedded Mouse cerebral cortex tissue labeling PPCEL/PREPL with ab203111 at 1/250 dilution followed by ab97051 Goat Anti-Rabbit IgG H&L (HRP) at a 1/500 dilution. Cytoplasmic staining on Mouse cerebral cortex tissue is observed. Counter stained with Hematoxylin.

Negative control : Used PBS instead of primary antibody secondary antibody is ab97051 Goat Anti-Rabbit IgG H&L (HRP).

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-PPCEL/PREPL antibody [EPR17752] (AB203111)

Immunohistochemical analysis of paraffin-embedded Rat cerebral cortex tissue labeling PPCEL/PREPL with ab203111 at 1/250 dilution followed by ab97051 Goat Anti-Rabbit IgG H&L (HRP) at a 1/500 dilution. Cytoplasmic staining on Rat cerebral cortex tissue is observed. Counter stained with Hematoxylin.

Negative control : Used PBS instead of primary antibody secondary antibody is ab97051 Goat Anti-Rabbit IgG H&L (HRP).

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

• IP

Supplier Data

Immunoprecipitation - Anti-PPCEL/PREPL antibody [EPR17752] (AB203111)

PPCEL/PREPL was immunoprecipitated from 1mg of NIH/3T3 (Mouse embyro fibroblast cells) whole cell extract with ab203111 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab203111 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

Lane 1 : NIH/3T3 whole cell extract 10 μg (Input). Lane 2 : ab203111 IP in NIH/3T3 whole cell extract. Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab203111 in NIH/3T3 whole cell extract.
Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 10 seconds

Blocking/Dilution Buffer : 5% NFDM/TBST

All lanes:

Immunoprecipitation - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)

Predicted band size: 84 kDa

false

• WB

Supplier Data

Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (AB203111)

Blocking/Dilution buffer 5% NFDM/TBST.

The observed MW is consistent with what has been described in the literature PMID : 16385448.

All lanes:

Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111) at 1/2000 dilution

Lane 1:

Human fetal brain lysate at 20 µg

Lane 2:

MCF-7 (Human breast adenocarcinoma cell line) whole cell lysate at 20 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 84 kDa

Observed band size: 72 kDa

false

Exposure time: 1min

• WB

Supplier Data

Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (AB203111)

Blocking/DIlution buffer 5% NFDM/TBST.

The observed MW is consistent with what has been described in the literature PMID : 16385448.

All lanes:

Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111) at 1/2000 dilution

Lane 1:

Mouse brain whole cell lysate at 10 µg

Lane 2:

Rat brain whole cell 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

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (AB203111)

Blocking/Dilution buffer 5% NFDM/TBST.

The observed MW is consistent with what has been described in the literature PMID : 16385448.

All lanes:

Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111) at 1/2000 dilution

Lane 1:

C6 (Rat glial tumor cells) whole cell lysate at 10 µg

Lane 2:

Raw264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus whole cell lysate at 10 µg

Lane 3:

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

Lane 4:

NIH/3T3 (Mouse embyro fibroblast cells) whole cell 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

false

Exposure time: 30s

• WB

Lab

Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (AB203111)

Exposure time :

Lane 1 : 3 seconds.

Lane 2 : 3 minutes.

Blocking and dilution buffer : 5% NFDM/TBST.

Lane 1:

Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111) at 1/1000 dilution

Lane 2:

Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111) at 1/10000 dilution

All lanes:

Recombinant Human PPCEL/PREPL protein (<a href='/en-us/products/unavailable/recombinant-human-ppcelprepl-protein-ab160557'>ab160557</a>) at 0.01 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 1/20000 dilution

Predicted band size: 84 kDa

Observed band size: 109 kDa

false