Name: Anti-Nanog antibody [EPR2027(2)]
Brand: RabMAb
SKU: ab109250
Rating: 4 (11 reviews)

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Western blot - Anti-Nanog antibody [EPR2027(2)] (ab109250)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR2027(2)] to Nanog
Suitable for: WB, IHC-P, ICC/IF, Flow Cyt (Intra), ChIP-sequencing, IP
Reacts with: Human

Alexa Fluor® 488 Alexa Fluor® 647 Carrier Free

产品名称

Anti-Nanog抗体[EPR2027(2)]
参阅全部 Nanog 一抗
描述

兔单克隆抗体[EPR2027(2)] to Nanog
宿主

Rabbit
特异性

100% identities with NANOGP8
经测试应用

适用于: WB, IHC-P, ICC/IF, Flow Cyt (Intra), ChIP-sequencing, IPmore details
种属反应性

与反应: Human
免疫原

Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
阳性对照
- WB: NCCIT cell lysate. IHC-P: Human seminoma tissue, Human dysgerminoma tissue and Human embryonal carcinoma tissue. ICC/IF: Human embryonic carcinoma and Human liver cell lines. IP: NCCIT whole cell lysate. ChIP-seq: NCCIT cells.
常规说明
Please note that Nanog is expressed variably in different tissues and that optimisation may be required depending on the tissue used for the experiment.
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.
Mouse: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

形式

Liquid
存放说明

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.
存储溶液

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 59% PBS, 0.05% BSA
Concentration information loading...
纯度

Protein A purified
克隆

单克隆
克隆编号

EPR2027(2)
同种型

IgG
研究领域

Alternative Versions
Compatible Secondaries
- Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
- Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant human Nanog protein (ab134886)
Related Products
- Prestained Protein Ladder - Broad molecular weight (10 - 245 kDa) (ab116028)

The Abpromise guarantee

Abpromise™承诺保证使用ab109250于以下的经测试应用

“应用说明”部分下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。

应用	Ab评论	说明
WB	(4)	1/1000 - 1/10000. Detects a band of approximately 37 kDa (predicted molecular weight: 35 kDa).
IHC-P	(3)	1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. Antigen retrieval is recommended.
ICC/IF	(4)	1/100 - 1/250.
Flow Cyt (Intra)		1/70. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ChIP-sequencing		Use at an assay dependent concentration.
IP		1/40.

说明
WB 1/1000 - 1/10000. Detects a band of approximately 37 kDa (predicted molecular weight: 35 kDa).
IHC-P 1/100 - 1/250. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. Antigen retrieval is recommended.
ICC/IF 1/100 - 1/250.
Flow Cyt (Intra) 1/70. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ChIP-sequencing Use at an assay dependent concentration.
IP 1/40.

功能

Transcription regulator involved in inner cell mass and embryonic stem (ES) cells proliferation and self-renewal. Imposes pluripotency on ES cells and prevents their differentiation towards extraembryonic endoderm and trophectoderm lineages. Blocks bone morphogenetic protein-induced mesoderm differentiation of ES cells by physically interacting with SMAD1 and interfering with the recruitment of coactivators to the active SMAD transcriptional complexes (By similarity). Acts as a transcriptional activator or repressor (By similarity). Binds optimally to the DNA consensus sequence 5'-TAAT[GT][GT]-3' or 5'-[CG][GA][CG]C[GC]ATTAN[GC]-3' (By similarity). When overexpressed, promotes cells to enter into S phase and proliferation.
组织特异性

Expressed in testicular carcinoma and derived germ cell tumors (at protein level). Expressed in fetal gonads, ovary and testis. Also expressed in ovary teratocarcinoma cell line and testicular embryonic carcinoma. Not expressed in many somatic organs and oocytes.
序列相似性

Belongs to the Nanog homeobox family.
Contains 1 homeobox DNA-binding domain.
发展阶段

Expressed in embryonic stem (ES) and carcinoma (EC) cells. Expressed in inner cell mass (ICM) of the blastocyst and gonocytes between 14 and 19 weeks of gestation (at protein level). Not expressed in oocytes, unfertilized oocytes, 2-16 cell embryos and early morula (at protein level). Expressed in embryonic stem cells (ES). Expression decreases with ES differentiation.
细胞定位

Nucleus.
Target information above from: UniProt accession Q9H9S0 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
数据库链接
- Entrez Gene: 100293888 Human
- Entrez Gene: 79923 Human
- Omim: 607937 Human
- SwissProt: Q9H9S0 Human
- Unigene: 635882 Human
别名
- Embryonic stem cell specific homeobox protein (Nanog) antibody
- ENK antibody
- FLJ12581 antibody
- hNanog antibody
- Homeobox protein NANOG antibody
- Homeobox transcription factor Nanog antibody
- homeobox transcription factor Nanog-delta 48 antibody
- NANOG antibody
- Nanog homeobox antibody
- NANOG_HUMAN antibody
see all

Western blot - Anti-Nanog antibody [EPR2027(2)] (ab109250)

All lanes : Anti-Nanog antibody [EPR2027(2)] (ab109250) at 1/2000 dilution

Lane 1 : NCCIT (Human pluripotent embryonic carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 2 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 3 : HEK-293 (Human embryonic kidney epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 4 : MDA-MB-231 (Human breast adenocarcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 5 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 6 : Huh7 (Human hepatocellular carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 7 : HCT 116 (Human colorectal carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 8 : PANC-1 (Human pancreatic epithelioid carcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST

Lysates/proteins at 20 µg per lane.

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

Predicted band size: 35 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?

Exposure time: 30 seconds

Nanog is highly expressed in cancer stem cells. Although some papers support the expression in undifferentiated cancer cell lines, such as HeLa (PMID: 22337995, 28092370), MDA-MB-231 (PMID: 28401007, 25919570), HepG2 (PMID: 29477378), Huh7 (PMID: 26919045), HCT 116 (PMID: 25249558, 28092370) and PANC-1 (PMID: 28703793, 25846752), ab109250 can’t detect the target band in these cell lines, even at the dilution of 1:200.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR2027(2)] (ab109250)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human seminoma tissue labelling Nanog with purified ab109250 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
Immunoprecipitation - Anti-Nanog antibody [EPR2027(2)] (ab109250)

ab109250 (purified) at 1/40 dilution (1.5 µg/ml) immunoprecipitating Nanog in NCCIT whole cell lysate.

Lane 1 (input): NCCIT(Human pluripotent embryonic carcinoma epithelial cell) whole cell lysate 10µg
Lane 2 (+): ab109250 & NCCIT whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab109250 in NCCIT whole cell lysate

For western blotting, ab109250 at 1/500 dilution (1.5 µg/ml) VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1/1000 dilution.

Blocking and diluting buffer: 5% NFDM /TBST.
Western blot - Anti-Nanog antibody [EPR2027(2)] (ab109250)

Anti-Nanog antibody [EPR2027(2)] (ab109250) at 1/1000 dilution (unpurified) + NCCIT cell lysate at 10 µg

Predicted band size: 35 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR2027(2)] (ab109250)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human seminoma tissue labelling Nanog with unpurified ab109250 at 1/100.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR2027(2)] (ab109250)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human dysgerminoma tissue labelling Nanog with unpurified ab109250 at 1/100.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR2027(2)] (ab109250)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human embryonal carcinoma tissue labelling Nanog with unpurified ab109250 at 1/100.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR2027(2)] (ab109250)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human normal colon tissue shows negative staining of Nanog with unpurified ab109250.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Nanog antibody [EPR2027(2)] (ab109250)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human adult kidney tissue shows negative staining of Nanog with unpurified ab109250.
Flow Cytometry (Intracellular) - Anti-Nanog antibody [EPR2027(2)] (ab109250)

Intracellular Flow Cytometry analysis ofNCCIT cells labelling Nanog with purified ab109250 at 1/70 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
Immunocytochemistry/ Immunofluorescence - Anti-Nanog antibody [EPR2027(2)] (ab109250)This image is courtesy of an anonymous Abreview

Immunocytochemistry/Immunofluorescence analysis of Human Liver cells labelling Nanog with unpurified ab109250. Cells were fixed with Paraformaldehyde, permeabilized with Triton X-100 0.1% and blocked with 1% BSA for 12 hours at 4°C. Sample was incubated with primary antibody (1/500 in PBS) for 16 hour at 4°C. An Alexa Fluor^®647-conjugated Donkey anti-rabbit(1/1000) IgG polyclonal was used as the secondary antibody.
See Abreview
Immunocytochemistry/ Immunofluorescence - Anti-Nanog antibody [EPR2027(2)] (ab109250)

Immunocytochemistry/Immunofluorescence analysis of embryonic carcinoma cells labelling Nanog with unpurified ab109250 at 1/100.
Immunocytochemistry/ Immunofluorescence - Anti-Nanog antibody [EPR2027(2)] (ab109250)

Immunocytochemistry/Immunofluorescence analysis of NCCIT(human pluripotent embryonal carcinoma) cells labelling Nanog with purified ab109250 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500).
ChIP-sequencing - Anti-Nanog antibody [EPR2027(2)] (ab109250)

Chromatin was prepared from NCCIT (Human pluripotent embryonic carcinoma cell line) cells. ChIP was performed with 10^7 NCCIT cells and 8 µg of ab109250 [[EPR2027(2)]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads.
Additional screenshots of mapped reads can be downloaded here.
Anti-Nanog antibody [EPR2027(2)] (ab109250)

Flow cytometry protocols
Immunohistochemistry protocols
Immunocytochemistry & immunofluorescence protocols
Western blot protocols
Protocol Booklet

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

发表研究结果有使用 ab109250？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab109250 被引用在 195 文献中.

Zhang W et al. Single-cell sequencing reveals SATB2/NOTCH1 signaling promotes the progression of malignancy of epithelial cells from papillary thyroid cancer. Mol Carcinog 63:22-33 (2024). PubMed: 37877736
Alowaysi M et al. Generation of iPSC lines (KAIMRCi003A, KAIMRCi003B) from a Saudi patient with Dravet syndrome carrying homozygous mutation in the CPLX1 gene and heterozygous mutation in SCN9A. Hum Cell 37:502-510 (2024). PubMed: 38110787
Lu X et al. Single-cell multi-omics analysis of human testicular germ cell tumor reveals its molecular features and microenvironment. Nat Commun 14:8462 (2023). PubMed: 38123589
Hao Q et al. Hsa_circRNA_001676 accelerates the proliferation, migration and stemness in colorectal cancer through regulating miR-556-3p/G3BP2 axis. Sci Rep 13:18353 (2023). PubMed: 37884630
Wang X et al. m6A modified BACE1-AS contributes to liver metastasis and stemness-like properties in colorectal cancer through TUFT1 dependent activation of Wnt signaling. J Exp Clin Cancer Res 42:306 (2023). PubMed: 37986103

View all Publications for this product

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1-4 of 4 Abreviews or Q&A

Application

Western blot

Sample

Human Cell lysate - whole cell (DU145 Prostate cancer cell line)

Gel Running Conditions

Reduced Denaturing

Loading amount

20 µg

Specification

DU145 Prostate cancer cell line

Blocking step

Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Feb 10 2020

Application

Western blot

Sample

Human Cell lysate - whole cell (patient derived iPSC)

Gel Running Conditions

Reduced Denaturing

Loading amount

10 µg

Specification

patient derived iPSC

Blocking step

Milk as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 5µg/mL · Temperature: 25°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Oct 05 2015

Application

Western blot

Sample

Human Cell lysate - whole cell (SW480 cells, HCT116 cells)

Loading amount

25 µg

Specification

SW480 cells, HCT116 cells

Gel Running Conditions

Reduced Denaturing (4-12%)

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Jan 17 2013

Application

Western blot

Sample

Human Cell lysate - whole cell (liver cancer)

Loading amount

20 µg

Specification

liver cancer

Gel Running Conditions

Reduced Denaturing (10%gel)

Blocking step

Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

提交于 Oct 18 2012

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Key features and details

Related conjugates and formulations

概述

产品名称

描述

宿主

特异性

经测试应用

种属反应性

免疫原

阳性对照

常规说明

性能

形式

存放说明

存储溶液

纯度

克隆

克隆编号

同种型

研究领域

相关产品

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

应用

The Abpromise guarantee

靶标

功能

组织特异性

序列相似性

发展阶段

细胞定位

数据库链接

别名

图片

实验方案

数据表及文件

SDS download

Datasheet download

文献 (195)

客户评价及客户问答

Western blot abreview for Anti-Nanog antibody [EPR2027(2)]

Western blot abreview for Anti-Nanog antibody [EPR2027(2)]

Western blot abreview for Anti-Nanog antibody [EPR2027(2)]

Western blot abreview for Anti-Nanog antibody [EPR2027(2)]