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

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

Suitable for: WB, IHC-P, ICC/IF, IPmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment within Human PPCEL/PREPL aa 600-700. The exact sequence is proprietary.
Database link: Q4J6C6
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.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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

Alternative Versions
- Anti-PPCEL/PREPL antibody [EPR17752] - BSA and Azide free (ab251393)
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab97051)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control - BSA and Azide Free (ab210849)

Our Abpromise guarantee covers the use of ab203111 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application	Abreviews	Notes
WB		1/2000. Detects a band of approximately 72 kDa (predicted molecular weight: 84 kDa).
IHC-P		1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF		1/250.
IP		1/30.

Function

Probable serine peptidase whose precise substrate specificity remains unclear. Does not cleave peptides after a arginine or lysine residue.
Tissue specificity

Widely expressed. Expressed at higher level in brain, skeletal muscle, heart and kidney.
Involvement in disease

Defects in PREPL are a cause of hypotonia-cystinuria syndrome (HCS) [MIM:606407]. HCS is characterized generalized hypotonia at birth, nephrolithiasis, growth hormone deficiency, minor facial dysmorphism, failure to thrive, followed by hyperphagia and rapid weight gain in late childhood. HCS is caused by a deletion that disrups both SLC3A1 and PREPL genes. As SLC3A1 is known to cause isolated cystinuria type I, the extended phenotype could be attributed to the deletion of PREPL.
Sequence similarities

Belongs to the peptidase S9A family.
Cellular localization

Cytoplasm > cytosol.
Target information above from: UniProt accession Q4J6C6 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 9581 Human
- Entrez Gene: 213760 Mouse
- Entrez Gene: 298771 Rat
- Omim: 609557 Human
- SwissProt: Q4J6C6 Human
- SwissProt: Q8C167 Mouse
- SwissProt: Q5HZA6 Rat
- Unigene: 444349 Human
- Unigene: 461481 Mouse
- Unigene: 154511 Rat
see all
Alternative names
- PPCEL_HUMAN antibody
- prepl antibody
- Prolyl endopeptidase-like antibody
- Prolylendopeptidase-like antibody
- putative prolyl oligopeptidase antibody
see all

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)

Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Datasheet

Publishing research using ab203111? Please let us know so that we can cite the reference in this datasheet.

ab203111 has not yet been referenced specifically in any publications.

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

Key features and details

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Applications

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Certificate of Compliance

References (0)

Customer reviews and Q&As