Recombinant Anti-PEG10/EDR antibody [EPR20051] KO Tested (ab215035)

Product name

Anti-PEG10/EDR antibody [EPR20051]
See all PEG10/EDR primary antibodies
Description

Rabbit monoclonal [EPR20051] to PEG10/EDR
Host species

Rabbit
Tested applications

Suitable for: Flow Cyt (Intra), ICC/IF, IP, IHC-P, WBmore details
Species reactivity

Reacts with: Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: HEK 293, HeLa and HepG2 whole cell lysates; Human testis lysate. IHC-P: Human placenta, liver cancer and testis tissues. ICC/IF: HepG2 cells. Flow Cyt (intra): HepG2 cells. IP: HepG2 whole cell lysate.
General notes
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: 7.2
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

EPR20051
Isotype

IgG
Research areas

Alternative Versions
- Anti-PEG10/EDR antibody [EPR20051] - BSA and Azide free (ab240392)
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)
KO cell lines
- Human PEG10 (EDR) knockout HeLa cell line (ab265925)
KO cell lysates
- Human PEG10 (EDR) knockout HeLa cell lysate (ab258103)
Recombinant Protein
- Recombinant Human PEG10/EDR protein (ab161530)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab215035 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
Flow Cyt (Intra)		1/500.
ICC/IF		1/500.
IP		1/30.
IHC-P		1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB		1/1000. Detects a band of approximately 100 kDa (predicted molecular weight: 30,80 kDa).

Notes
Flow Cyt (Intra) 1/500.
ICC/IF 1/500.
IP 1/30.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 100 kDa (predicted molecular weight: 30,80 kDa).

Function

Prevents apoptosis in hepatocellular carcinoma (HCC) cells through interaction with SIAH1, a mediator of apoptosis. May also have a role in cell growth promotion and hepatoma formation. Inhibits the TGF-beta signaling by interacting with the TGF-beta receptor ALK1. When overexpressed, induces the formation of cellular extension, such as filipodia in association with ALK1. Involved at the immediate early stage of adipocyte differentiation (By similarity). May bind to the 5'-GCCTGTCTTT-3' DNA sequence of the MB1 domain in the myelin basic protein (MBP) promoter.
Tissue specificity

Expressed in the cytotrophoblast layer but not in the overlying syncytiotrophoblast of the placenta. Expressed in prostate and breast carcinomas but not in normal breast and prostate epithelial cells. Expressed in the HepG2 cell line (at protein level). Expressed in brain, liver, spleen, kidney, thymus, lung, ovary, testis, reactive lymph node, skeletal muscle, adipose tissue and placenta. Expressed in pancreatic and hepatocellular carcinomas (HCC).
Sequence similarities

Contains 1 CCHC-type zinc finger.
Developmental stage

Expressed in placenta during the first trimester of gestation (at protein level). In placenta, down-regulated at early hypoxic phase, and highly activated at 11-12 week of gestation.
Post-translational
modifications

Isoform RF1/RF2 undergoes proteolytic cleavage.
Cellular localization

Nucleus. Cytoplasm. Detected predominantly in the cytoplasm of breast and prostate carcinomas, in hepatocellular carcinoma (HCC) and B-cell chronic lymphocytic leukemia (B-CLL) cells and in the HepG2 cell line. Colocalized with ALK1.
Target information above from: UniProt accession Q86TG7 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
Database links
- Entrez Gene: 23089 Human
- Omim: 609810 Human
- SwissProt: Q86TG7 Human
- Unigene: 147492 Human
Alternative names
- AA407948 antibody
- Edr antibody
- Embryonal carcinoma differentiation regulated antibody
- Embryonal carcinoma differentiation-regulated protein antibody
- HB 1 antibody
- HB1 antibody
- KIAA1051 antibody
- Mammalian retrotransposon-derived protein 2 antibody
- Mar2 antibody
- Mart2 antibody
- MEF3 like 1 antibody
- MEF3-like protein 1 antibody
- MEF3L antibody
- MEF3L1 antibody
- MyEF 3 antibody
- Myelin expression factor 3-like protein 1 antibody
- Paternally expressed 10 antibody
- Paternally expressed gene 10 ORF1 antibody
- Paternally expressed gene 10 protein antibody
- Peg10 antibody
- PEG10 protein antibody
- PEG10_HUMAN antibody
- Putative uncharacterized protein PEG10 antibody
- Retrotransposon gag domain containing 3 antibody
- Retrotransposon gag domain-containing protein 3 antibody
- Retrotransposon-derived gag-like polyprotein antibody
- Retrotransposon-derived protein PEG10 antibody
- RGAG3 antibody
- Ty3/Gypsy-like protein antibody
see all

Western blot - Anti-PEG10/EDR antibody [EPR20051] (ab215035)

All lanes : Anti-PEG10/EDR antibody [EPR20051] (ab215035) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : PEG10 knockout HeLa cell lysate
Lane 3 : HepG2 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 30,80 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?

Lanes 1-4: Merged signal (red and green). Green - ab215035 observed at 100 kDa. Red - loading control ab7291 observed at 50 kDa.

ab215035 Anti-PEG10/EDR antibody [EPR20051] was shown to specifically react with PEG10/EDR in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265925 (knockout cell lysate ab258103) was used. Wild-type and PEG10/EDR knockout samples were subjected to SDS-PAGE. ab215035 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PEG10/EDR antibody [EPR20051] (ab215035)

Immunohistochemical analysis of paraffin-embedded human liver cancer tissue labeling PEG10/EDR with ab215035 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution. Cytoplasmic staining on part of the cells in human liver cancer is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunocytochemistry/ Immunofluorescence - Anti-PEG10/EDR antibody [EPR20051] (ab215035)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling PEG10/EDR with ab215035 at 1/500 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasmic staining on HepG2 cell line.

The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 (Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594)) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor^® 488) (ab150077) at 1/1000 dilution.
Western blot - Anti-PEG10/EDR antibody [EPR20051] (ab215035)

Lanes 1-2 : Anti-PEG10/EDR antibody [EPR20051] (ab215035) at 1/2000 dilution
Lane 3 : Anti-PEG10/EDR antibody [EPR20051] (ab215035) at 1/10000 dilution

Lane 1 : HEK 293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 30,80 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?

Exposure time: 3 minutes

Blocking/Dilution buffer: 5% NFDM/TBST.

The expression of PEG10/EDR in HepG2 cells is consistent with what has been described in the literature PMID: 17942406.
Flow Cytometry (Intracellular) - Anti-PEG10/EDR antibody [EPR20051] (ab215035)

Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling PEG10/EDR with ab215035 at 1/500 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (Alexa Fluor^® 488) at 1/2000 dilution was used as the secondary antibody.
Western blot - Anti-PEG10/EDR antibody [EPR20051] (ab215035)

Anti-PEG10/EDR antibody [EPR20051] (ab215035) at 1/1000 dilution + Human testis lysate at 10 µg

Secondary
Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/100000 dilution

Predicted band size: 30,80 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?

Exposure time: 1 minute

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PEG10/EDR antibody [EPR20051] (ab215035)

Immunohistochemical analysis of paraffin-embedded human placenta tissue labeling PEG10/EDR with ab215035 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution. Cytoplasmic staining on some cells in human placenta is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

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-PEG10/EDR antibody [EPR20051] (ab215035)

Immunohistochemical analysis of paraffin-embedded human testis tissue labeling PEG10/EDR with ab215035 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution. Cytoplasmic staining on the germ cells in human testis is observed. Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

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-PEG10/EDR antibody [EPR20051] (ab215035)

Immunohistochemical analysis of paraffin-embedded human liver tissue labeling PEG10/EDR with ab215035 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.

Negative staining on normal cells in human liver.

Counter stained with Hematoxylin.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunoprecipitation - Anti-PEG10/EDR antibody [EPR20051] (ab215035)

PEG10/EDR was immunoprecipitated from 0.35 mg of HepG2 (Human liver hepatocellular carcinoma cell line) whole cell lysate with ab215035 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab215035 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution

Lane 1: HepG2 whole cell lysate, 10 μg (Input).

Lane 2: ab215035 IP in HepG2 whole cell lysate.

Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab215035 in HepG2 whole cell lysate.

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

Exposure time: 3 seconds.
Anti-PEG10/EDR antibody [EPR20051] (ab215035)

Flow cytometry protocols
Immunoprecipitation protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols

Click here to view the general protocols

Datasheet download

Download

Publishing research using ab215035? Please let us know so that we can cite the reference in this datasheet.

ab215035 has been referenced in 2 publications.

Ding F et al. LncRNA MIR7-3HG executes a positive role in retinoblastoma progression via modulating miR-27a-3p/PEG10 axis. Exp Eye Res 193:107960 (2020). PubMed: 32035086
Jankovic-Karasoulos T et al. Isolation of villous cytotrophoblasts from second trimester human placentas. Placenta 74:55-58 (2018). PubMed: 30541665

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Key features and details

Related conjugates and formulations

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

KO cell lines

KO cell lysates

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Sequence similarities

Developmental stage

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

Datasheet download

Certificate of Compliance

References (2)

Customer reviews and Q&As

Post-translational
modifications