Antibodies, Proteins, Kits and Reagents for Life Science

Product name

Anti-PPCEL/PREPL antibody [EPR17752]
See all PPCEL/PREPL primary antibodies
Description

Rabbit monoclonal [EPR17752] to PPCEL/PREPL
Host species

Rabbit

Tested Applications & Species

Application	Species
ICC/IF	Human
IHC-P	Mouse Rat Human
IP	Mouse
WB	Mouse Rat Human

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: Human fetal brain, MCF-7, Mouse brain, Rat brain, C6, Raw264.7, PC-12 or NIH/3T3 whole cell lysate. IHC-P: Human, Mouse or Rat cerebral cortex tissue. ICC/IF: SH-SY5Y or MCF-7 cells. Flow Cyt: MCF-7 cells.
General notes
This product was previously labelled as PPCEL

This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR17752
Isotype

IgG
Research areas

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

Lane 1 : Anti-PPCEL/PREPL antibody [EPR17752] (ab203111) at 1/1000 dilution
Lane 2 : Anti-GST antibody [EPR4236] (ab111947) at 1/10000 dilution

All lanes : Recombinant Human PPCEL/PREPL protein (ab160557)

Lysates/proteins at 0.01 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 84 kDa
Observed band size: 109 kDa
why is the actual band size different from the predicted?

Exposure time:
Lane 1: 3 seconds.
Lane 2: 3 minutes.

Blocking and dilution buffer: 5% NFDM/TBST.
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.
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.
Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)

All lanes : Anti-PPCEL/PREPL antibody [EPR17752] (ab203111) at 1/2000 dilution

Lane 1 : Human fetal brain lysate
Lane 2 : MCF-7 (Human breast adenocarcinoma cell line) whole cell lysate

Lysates/proteins at 20 µg per lane.

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 why is the actual band size different from the predicted?

Exposure time: 1 minute

Blocking/Dilution buffer 5% NFDM/TBST.

The observed MW is consistent with what has been described in the literature PMID: 16385448.
Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)

All lanes : Anti-PPCEL/PREPL antibody [EPR17752] (ab203111) at 1/2000 dilution

Lane 1 : Mouse brain whole cell lysate
Lane 2 : Rat brain whole cell lysate

Lysates/proteins at 10 µg per lane.

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 why is the actual band size different from the predicted?

Exposure time: 15 seconds

Blocking/DIlution buffer 5% NFDM/TBST.

The observed MW is consistent with what has been described in the literature PMID: 16385448.
Western blot - Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)

All lanes : Anti-PPCEL/PREPL antibody [EPR17752] (ab203111) at 1/2000 dilution

Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
Lane 2 : Raw264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
Lane 4 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

Lysates/proteins at 10 µg per lane.

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 why is the actual band size different from the predicted?

Exposure time: 30 seconds

Blocking/Dilution buffer 5% NFDM/TBST.

The observed MW is consistent with what has been described in the literature PMID: 16385448.
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.
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.
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.
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
Anti-PPCEL/PREPL antibody [EPR17752] (ab203111)