Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Mouse

Cited:

Human, Mouse, Transgenic Mouse

Applications

Validated:

Immunohistochemistry, Western Blot, Immunocytochemistry, Chromatin Immunoprecipitation (ChIP)

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Flow Cytometry, Immunocytochemistry, Immunocytochemistry/ Immunofluorescence, Immunoprecipitation, Chromatin Immunoprecipitation (ChIP), Co-Immunoprecipitation, In Situ Hybridization

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all KLF4 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant mouse KLF4
Ala11-Phe483
Accession # Q60793

Specificity

Detects mouse KLF4 in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Mouse KLF4 Antibody

Detection of KLF4-regulated Genes antibody by Chromatin Immunoprecipitation.

Detection of KLF4-regulated Genes by Chromatin Immunoprecipitation.

D3 mouse embryonic stem cell line was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. KLF4/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Mouse KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3158) or control antibody (AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (BAF109). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (MAG999) and DNA was purified using chelating resin solution. TheB2Rpromoter was detected by standard PCR.

KLF4 antibody in D3 Mouse Cell Line by Immunocytochemistry (ICC).

KLF4 in D3 Mouse Cell Line.

KLF4 was detected in immersion fixed D3 mouse embryonic stem cell line using Mouse KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3158) at 10 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

KLF4 antibody in Mouse Colon Tissue by Immunohistochemistry (IHC-Fr).

KLF4 in Mouse Colon Tissue.

KLF4 was detected in perfusion fixed frozen sections of mouse colon tissue using Goat Anti-Mouse KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3158) at 1 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (CTS013). Tissue was stained using the Anti-Goat IgG VisUCyte™ HRP Polymer Antibody (brown; VC004) and counterstained with hematoxylin (blue). Specific staining was localized to cell nuclei. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.

Detection of Mouse KLF4 by Immunohistochemistry

Detection of Mouse KLF4 by Immunohistochemistry

Overexpression of beta -catenin maintains B6 ESC self-renewal in the presence of LIF.(A) Immunoblot analysis of phospho-beta -catenin, beta -catenin and beta -actin levels in B6 ES cells cultured in 1000 U/ml LIF condition supplemented without or with 3 µM CH for 4 days. (B) Immunofluorescence analysis of 3 µM CH-treated and non-treated B6 ESC clones stained for beta -catenin and Hoechst for nuclei in the presence of LIF for 4 days. Scale bars represent 100 µm. (C) Quantitative RT-PCR analysis of Axin2, T and Cdx1 transcript levels in 3 µM CH-treated and control B6 ES cells. Error bars represent the SD of three biological replicates. (D) beta -catenin S33A protein stably expressed in ES cells was detected by immunoblotting. (E) Phase contrast microscopy image and alkaline phosphatase staining of control and beta -catenin S33A-transfected B6 ES cells in the presence of 1000 U/ml LIF for 4 days. Scale bars represent 100 µm. (F) Immunofluorescence staining for OCT4, NANOG, and KLF4 in mock and beta -catenin S33A-transfected B6 ES cells in the presence of 1000 U/ml LIF for 4 days. Cell nuclei were stained with Hoechst. Scale bars represent 100 µm. (G) Quantitative RT-PCR analysis of Oct4, Nanog, Rex1, Klf4, Tbx3, and Fgf5 transcript levels in mock and beta -catenin S33A-transfected B6 ES cells in the presence of 1000 U/ml LIF for 4 days. Error bars represent the SD of three biological replicates. (H) beta -catenin S33A delta C, beta -catenin S33A and TCF4 delta N protein were detected by immunoblotting with anti-flag tag antibody from extracts of ES cells transfected with PiggyBac plasmids. Quantitative RT-PCR analysis of Axin2, T and Cdx1 transcript levels in beta -catenin S33A delta C, beta -catenin S33A, TCF4 delta N, and mock-transfected B6 ES cells. Error bars represent the SD of three biological replicates. (I) Alkaline phosphatase staining of mock-transfected, and beta -catenin delta C over-expressing B6 ES cells cultured in LIF for 5 passages, and mock-transfected and TCF4 delta N-transfected B6 ESCs cultured in CHIR99021 plus LIF for 5 passages. Scale bars represent 100 µm. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0035892), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse KLF4 by Immunocytochemistry/Immunofluorescence

Detection of Mouse KLF4 by Immunocytochemistry/Immunofluorescence

LIF maintains B6 ES cell self-renewal in the presence of CHIR99021.(A) Phase contrast image of B6 ES cells in the presence of 1000 U/ml LIF supplemented without or with 3 µM CHIR99021 (CH) for 4 days. Scale bars represent 100 µm. (B) Alkaline phosphatase staining of B6 ES cells in the presence of 1000 U/ml LIF supplemented without or with 3 µM CH for 4 days. Scale bars represent 100 µm. (C) Immunofluorescence staining for OCT4, NANOG, and KLF4 in B6 ES cells treated with 1000 U/ml LIF and 3 µM CH for 4 days. Cell nuclei were stained using Hoechst. Scale bars represent 100 µm. (D) Relative quantification of Oct4, Nanog, Rex1, Klf4, Tbx3, and Fgf5 mRNA in B6 ES cells in 1000 U/ml LIF culture without or with 3 µM CH, by qRT-PCR. Error bars represent the SD of three biological replicates. (E) B6 ES cells,1×104/well, cultured in different concentrations of LIF, ranging from 0 to 10,000 U/ml, in the absence or presence of 3 µM CH for 5 days, were assessed for alkaline phosphatase activity. Scale bars represent 100 µm. Image collected and cropped by CiteAb from the following publication (https://dx.plos.org/10.1371/journal.pone.0035892), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Mouse KLF4 by Chromatin Immunoprecipitation

Detection of Mouse KLF4 by Chromatin Immunoprecipitation

Overlapping SIM2 occupancy with master transcription factor binding sites.a. Frequency distribution of OCT4, SOX2 and NANOG DNA binding sites in a 40kb window centered to the newly identified SIM2 DNA binding sites. Plots show a significant enrichment for the OSN binding sites at the SIM2 peak localization in SIM2 A6 expressing cells. Pie charts show the proportion of SIM2 DNA binding sites overlapping with the OCT4, SOX2 or NANOG binding sites (in grey) (100bp window). p = Fisher’s exact test p-value; F score: measure of the significance of the association (1 = perfect match). b. Protein co-immunoprecipitation experiments of SIM2-FLAG with endogenous OCT4, SOX2, KLF4 (left panel) and NANOG (right panel). Cellular protein extracts from Sim2 expressing cells (A6) or EB3 cells were immunoprecipitated by using antibodies directed against each of the pluripotency factors (N-terminal and C-terminal part of NANOG) or IgG as a negative control for co-immunoprecipitation. Associated proteins were immunoblotted using an anti-FLAG antibody. Red star shows the SIM2-FLAG protein, blue star the signal given by the recognition of the IgG heavy chains. Ø: Beads only; kDa: kilodaltons; protein lysat: protein lysat was loaded as an input control for the immunoblot. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/25955728), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of KLF4 by Western Blot

Detection of KLF4 by Western Blot

CMTR1 is required for histone and ribosomal protein gene expression during differentiation. (A) Cmtr1WT,Cmtr1KD and Cmtr1KD/HA-CMTR1 ESCs were cultured in 35S cysteine/methionine for 30 min and incorporation into cellular proteins was determined (N = 3). Average and standard deviation is presented. Student’s T test; ‘*’ indicates P-value < 0.05 relative to control. (B,C) Western blot analysis of ESC cultured in (B) LIF and (C) without LIF for 2 days following CMTR1 siRNA transfection; N = 3. (D) Cmtr1WT and Cmtr1KD ESC were cultured in and without LIF for the days indicated. Cell number was counted; N = 3. Average and standard deviation presented. Student’s T test; ‘*’ indicates P-value < 0.05 relative to control timepoint. (E) Cmtr1WT and Cmtr1KDESC were induced to differentiate (E) by LIF withdrawal, (F) towards a neural fate. Phase contrast images and representative western blot analysis. (G) The ESC lines indicated were cultured without LIF for 6 days, then stained with alkaline phosphatase. Resultant colonies were scored as alkaline phosphatase (AP) negative (-), positive (+) or mixed; N = 4. Average and standard deviation presented. Student’s T test performed on data from AP- colonies; ‘*’ indicates P-value < 0.05 relative to control timepoint. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/35212377), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Mouse KLF4 Antibody

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

5 µg/5 x 106 cells
Sample: D3 mouse embryonic stem cell line chromatin, B2R promoter detected by standard PCR.

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed D3 mouse embryonic stem cell line

Immunohistochemistry

1-25 µg/mL
Sample: Perfusion fixed frozen sections of mouse colon tissue

Western Blot

0.1 µg/mL
Sample: Recombinant Mouse KLF4

Reviewed Applications

Read 4 reviews rated 4.5 using AF3158 in the following applications:

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

Stability & Storage

Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: KLF4

KLF4 belongs to the KLF family of transcription factors that regulate several cell functions. It is highly expressed in differentiated epithelial cells of the gastrointestinal tract. Human and mouse KLF4 share 91% amino acid sequence homology.

Long Name

Kruppel-Like Factor 4

Alternate Names

EZF

Entrez Gene IDs

9314 (Human)

Gene Symbol

KLF4

UniProt

Q60793

Additional KLF4 Products

Product Documents for Mouse KLF4 Antibody

Certificate of Analysis

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Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Mouse KLF4 Antibody

For research use only

Citations for Mouse KLF4 Antibody

Customer Reviews for Mouse KLF4 Antibody (4)

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Customer Images

Showing  1 - 4 of 4 reviews Showing All
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  • Mouse KLF4 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Mouse intestinal tissue, Human colorectal cancer tissue, HCT116 , Mouse embryonic fibroblasts and Caco-2 human colorectal adenocarcinoma cell line
    Species: Mouse and Human
    Verified Customer | Posted 02/22/2016
  • Mouse KLF4 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Mouse embryonic fibroblasts
    Species: Mouse
    Verified Customer | Posted 10/26/2015
    MEFs expressing indicated constructs were lysed and total lysates run on 10% gels.<br /> Blots were blocked in 5% milk in TBST, incubated with antibody in 1% milk in TBST over night at 4C, washed 3x in TBST for 15min, incubated with HRP-conjugated secondary antibody in 1% milk in TBST for 1h, washed again 3x in TBST for 15min and then developed.<br /> Bands in other than Klf4 lane show endogenous Klf4 proteins. <br />Specificity: Specific<br />Sensitivity: Sensitive<br />Buffer: TBST + 1% milk<br />Dilution: 1:1000
    Mouse KLF4 Antibody AF3158
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 23395636
    Species: Mouse
    Verified Customer | Posted 01/07/2015
  • Name: Anonymous
    Application: Western Blot
    Sample Tested: See PMID 20550931
    Species: Mouse
    Verified Customer | Posted 01/07/2015

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Protocols

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FAQs for Mouse KLF4 Antibody

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  • Q: Can I use this antibody in rabbit samples?

    A: AF3158 is against KLF4 and is derived from mouse. Unfortunately, it has not been tested with the rabbit samples. Therefore we don't know for sure whether this antibody will work in rabbit. Since it is a polyclonal antibody, the likelihood that this antibody will recognize the rabbit KLF4 is good.

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