Recombinant Anti-Nanog antibody [EPR2027(2)] - BSA and Azide free (ab218524)
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 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, ChIP-sequencing, IP
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Nanog antibody [EPR2027(2)] - BSA and Azide free
See all Nanog primary antibodies -
Description
Rabbit monoclonal [EPR2027(2)] to Nanog - BSA and Azide free -
Host species
Rabbit -
Specificity
100% identities with NANOGP8
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Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF, ChIP-sequencing, IPmore details -
Species reactivity
Reacts with: Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- 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.
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General notes
ab218524 is the carrier-free version of ab109250.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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
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.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.20
Constituent: PBS -
Carrier free
Yes -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR2027(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab218524 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 |
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
Use at an assay dependent concentration. Detects a band of approximately 37 kDa (predicted molecular weight: 35 kDa).
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IHC-P |
Use at an assay dependent concentration. 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. |
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ICC/IF |
Use at an assay dependent concentration.
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ChIP-sequencing |
Use at an assay dependent concentration.
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IP |
Use at an assay dependent concentration.
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Notes |
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
Use at an assay dependent concentration. Detects a band of approximately 37 kDa (predicted molecular weight: 35 kDa). |
IHC-P
Use at an assay dependent concentration. 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
Use at an assay dependent concentration. |
ChIP-sequencing
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
Target
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Function
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. -
Tissue specificity
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. -
Sequence similarities
Belongs to the Nanog homeobox family.
Contains 1 homeobox DNA-binding domain. -
Developmental stage
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. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 100293888 Human
- Entrez Gene: 79923 Human
- Omim: 607937 Human
- SwissProt: Q9H9S0 Human
- Unigene: 635882 Human
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Alternative names
- Embryonic stem cell specific homeobox protein (Nanog) antibody
- ENK antibody
- FLJ12581 antibody
see all
Images
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109250).
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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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109250).
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Intracellular Flow Cytometry analysis of NCCIT 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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109250).
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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 lysateFor 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human adult kidney tissue shows negative staining of Nanog with unpurified ab109250.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109250).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human seminoma tissue labelling Nanog with unpurified ab109250 at 1/100.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109250).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human dysgerminoma tissue labelling Nanog with unpurified ab109250 at 1/100.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109250).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human embryonal carcinoma tissue labelling Nanog with unpurified ab109250 at 1/100.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109250).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human normal colon tissue shows negative staining of Nanog with unpurified ab109250.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109250).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109250).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab109250).
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (15)
ab218524 has been referenced in 15 publications.
- He Y et al. CircZNF609 enhances hepatocellular carcinoma cell proliferation, metastasis, and stemness by activating the Hedgehog pathway through the regulation of miR-15a-5p/15b-5p and GLI2 expressions. Cell Death Dis 11:358 (2020). PubMed: 32398664
- Zhang L et al. CircAGFG1 drives metastasis and stemness in colorectal cancer by modulating YY1/CTNNB1. Cell Death Dis 11:542 (2020). PubMed: 32681092
- Chen L et al. Silencing transcription factor FOXM1 represses proliferation, migration, and invasion while inducing apoptosis of liver cancer stem cells by regulating the expression of ALDH2. IUBMB Life 72:285-295 (2020). PubMed: 31580537
- Zhao H et al. Exosomes from CD133+ cells carrying circ-ABCC1 mediate cell stemness and metastasis in colorectal cancer. J Cell Biochem 121:3286-3297 (2020). PubMed: 31960989
- Hu Q et al. MicroRNA-137 exerts protective effects on hypoxia-induced cell injury by inhibiting autophagy/mitophagy and maintaining mitochondrial function in breast cancer stem-like cells. Oncol Rep 44:1627-1637 (2020). PubMed: 32945512