Recombinant Anti-KLF4 antibody [EPR19590] (ab215036)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19590] to KLF4
- Suitable for: IP, Flow Cyt (Intra), IHC-P, WB, ICC/IF
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-KLF4 antibody [EPR19590]
See all KLF4 primary antibodies -
Description
Rabbit monoclonal [EPR19590] to KLF4 -
Host species
Rabbit -
Tested applications
Suitable for: IP, Flow Cyt (Intra), IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: NCCIT whole cell lysate. IHC-P: Human stomach, gastric cancer and seminoma tissues. ICC/IF: NCCIT cells. Flow Cyt (intra): NCCIT cells. IP: NCCIT cell lysate
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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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, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19590 -
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
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab215036 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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IP |
1/30.
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Flow Cyt (Intra) |
1/50.
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IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB | (1) |
1/1000. Detects a band of approximately 54 kDa (predicted molecular weight: 54 kDa).
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ICC/IF | (3) |
1/100.
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Notes |
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IP
1/30. |
Flow Cyt (Intra)
1/50. |
IHC-P
1/2000. 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 54 kDa (predicted molecular weight: 54 kDa). |
ICC/IF
1/100. |
Target
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Function
Transcription factor; can act both as activator and as repressor. Binds the 5'-CACCC-3' core sequence. Binds to the promoter region of its own gene and can activate its own transcription. Regulates the expression of key transcription factors during embryonic development. Plays an important role in maintaining embryonic stem cells, and in preventing their differentiation. Required for establishing the barrier function of the skin and for postnatal maturation and maintenance of the ocular surface. Involved in the differentiation of epithelial cells and may also function in skeletal and kidney development. Contributes to the down-regulation of p53/TP53 transcription. -
Sequence similarities
Belongs to the krueppel C2H2-type zinc-finger protein family.
Contains 3 C2H2-type zinc fingers. -
Domain
the 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors. -
Cellular localization
Nucleus. - Information by UniProt
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Database links
- Entrez Gene: 9314 Human
- Omim: 602253 Human
- SwissProt: O43474 Human
- Unigene: 376206 Human
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Alternative names
- Endothelial Kruppel like zinc finger protein antibody
- Epithelial zinc finger protein EZF antibody
- EZF antibody
see all
Images
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Anti-KLF4 antibody [EPR19590] (ab215036) at 1/1000 dilution + NCCIT (Human pluripotent embryonic carcinoma cell line) whole cell lysate at 10 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Predicted band size: 54 kDa
Observed band size: 54 kDa
Exposure time: 10 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NCCIT (Human pluripotent embryonic carcinoma cell line) cells labeling KLF4 with ab215036 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing nuclear staining on NCCIT cell line.
The nuclear counterstain 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KLF4 antibody [EPR19590] (ab215036)
Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling KLF4 with ab215036 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.
Nuclear staining on human stomach 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.
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ab215036 (purified) at 1/30 dilution immunoprecipitating KLF4 in NCCIT whole cell lysate.
Lane 1 (input): NCCIT (Human pluripotent embryonic carcinoma epithelial cell) whole cell lysate 10 µg
Lane 2 (+): ab215036 & NCCIT whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab215036 in NCCIT whole cell lysate
For western blotting, ab215036 at 1/500 dilution (0.117 µg/ml) and veriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/1000 dilution were used.
Blocking and diluting buffer: 5% NFDM /TBST . -
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed NCCIT (Human pluripotent embryonic carcinoma cell line) cells labeling KLF4 with ab215036 at 1/50 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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KLF4 antibody [EPR19590] (ab215036)
Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling KLF4 with ab215036 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.
Nuclear staining on tumor cells of human gastric cancer is observed [PMID: 17614846].
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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KLF4 antibody [EPR19590] (ab215036)
Immunohistochemical analysis of paraffin-embedded human seminoma tissue labeling KLF4 with ab215036 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) at 1/500 dilution.
Nuclear staining on tumor cells of human seminoma is observed [PMID: 17614846].
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.
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All lanes : Anti-KLF4 antibody [EPR19590] (ab215036) at 1/1000 dilution
Lane 1 : GST tagged Recombinant Human KLF4 protein (Full length, 81 KDa)
Lane 2 : GST tagged Recombinant Human Adipose Triglyceride Lipase (PNPLA2) protein (Full length, 82 KDa)
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 54 kDa
Observed band size: 81 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondBlocking buffer and concentration: 5% NFDM/TBST
Diluting buffer and concentration: 5% NFDM/TBST
Gel type: 4-20% gradient gel(SDS-PAGE)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (40)
ab215036 has been referenced in 40 publications.
- Chen C et al. Translational and post-translational control of human naïve versus primed pluripotency. iScience 25:103645 (2022). PubMed: 35005567
- Sun Y et al. Involvement of Cancer Stem Cells in Chemoresistant Relapse of Epithelial Ovarian Cancer Identified by Transcriptome Analysis. J Oncol 2022:6406122 (2022). PubMed: 35401749
- Fei Z et al. Krüppel-like factor 4 promotes the proliferation and osteogenic differentiation of BMSCs through SOX2/IGF2 pathway. Acta Biochim Pol 69:349-355 (2022). PubMed: 35617351
- Cao C et al. GCN5 participates in KLF4-VEGFA feedback to promote endometrial angiogenesis. iScience 25:104509 (2022). PubMed: 35733790
- Liang J et al. Saikosaponin D improves chemosensitivity of glioblastoma by reducing the its stemness maintenance. Biochem Biophys Rep 32:101342 (2022). PubMed: 36186734