Key Product Details

Validated by

Knockout/Knockdown

Species Reactivity

Validated:

Human

Cited:

Human, Mouse, Transgenic Mouse

Applications

Validated:

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

Cited:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry, Immunoprecipitation, Chromatin Immunoprecipitation (ChIP), Co-Immunoprecipitation, Mobility Shift Assay

Label

Unconjugated

Antibody Source

Polyclonal Goat IgG
View all KLF4 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human KLF4
Ala2-Phe470
Accession # AAH29923

Specificity

Detects human KLF4 in direct ELISAs and Western blots.

Clonality

Polyclonal

Host

Goat

Isotype

IgG

Scientific Data Images for Human KLF4 Antibody

Detection of Human KLF4 antibody by Western Blot.

Detection of Human KLF4 by Western Blot.

Western blot shows lysates of HT-29 human colon adenocarcinoma cell line, SW480 human colorectal adenocarcinoma cell line, and HCT-116 human colorectal carcinoma cell line. PVDF membrane was probed with 0.5 µg/mL of Goat Anti-Human KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3640) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). A specific band was detected for KLF4 at approximately 60 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 5.

KLF4 antibody in BG01V Human Stem Cells by Immunocytochemistry (ICC).

KLF4 in BG01V Human Stem Cells.

KLF4 was detected in immersion fixed BG01V human embryonic stem cells using 10 µg/mL Goat Anti-Human KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3640) for 3 hours at room temperature. Cells were stained with 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.

Detection of KLF4-regulated Genes antibody by Chromatin Immunoprecipitation.

Detection of KLF4-regulated Genes by Chromatin Immunoprecipitation.

BG01V human embryonic stem cells were fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. KLF4/DNA complexes were immunoprecipitated using 5 µg Goat Anti-Human KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3640) 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 Human Colon by Immunohistochemistry (IHC-P).

KLF4 in Human Colon.

KLF4 was detected in immersion fixed paraffin-embedded sections of human colon using 15 µg/mL Goat Anti-Human KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3640) overnight at 4 °C. Tissue was stained with the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). Specific labeling was localized to the nuclei of epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Detection of Human KLF4 antibody by Simple WesternTM.

Detection of Human KLF4 by Simple WesternTM.

Simple Western lane view shows lysates of HT-29 human colon adenocarcinoma cell line, loaded at 0.2 mg/mL. A specific band was detected for KLF4 at approximately 63 kDa (as indicated) using 25 µg/mL of Goat Anti-Human KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3640) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.

Detection of KLF4 in HT‑29 Human Colon Adenocarcinoma Cell Line (positive) and HDLM‑2 Human Hodgkin’s Lymphoma Cell Line (negative) Cells.

KLF4 was detected in immersion fixed HT‑29 Human Colon Adenocarcinoma Cell Line (positive) and HDLM‑2 Human Hodgkin’s Lymphoma Cell Line (negative) Cells using Goat Anti-Human KLF4 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3640) at 5 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Goat IgG Secondary Antibody (red; Catalog # NL001) and counterstained with DAPI (blue). Specific staining was localized to cytoplasm. View our protocol for Fluorescent ICC Staining of Cells on Coverslips.

Detection of Mouse KLF4 by Knockdown Validated

Detection of Mouse KLF4 by Knockdown Validated

KLF4 regulates the EndMT switch in CCM1 KO ECsA–DCultured lung‐derived WT and CCM1 KO ECs were lentivirally transduced with shRNA directed to either Klf4 (shKLF4) or control sequence (shCTRL). (A) qRT–PCR of mesenchymal (Fsp1, Id1), stem cell‐like (Sca1), and endothelial markers (VE‐cadherin and Claudin5) in WT shCTRL, WT shKLF4, CCM1 KO shCTRL, and CCM1 KO shKLF4 ECs. Data are mean ± SD (n = 3). Fold difference in gene expression is relative to WT shCTRL ECs. A two‐tailed unpaired t‐test was performed. Klf4: ****P < 0.00001, ***P = 0.0008, **P = 0.007; Fsp1: ***P = 0.0001, ****P = 3.9E‐05, ###P = 0.0002; Sca1: ***P = 0.0001, ****P < 0.00001; Id1: ***P = 0.0001; Ve‐cadherin: ***P = 0.0001; Claudin5: ****P = 1.37E‐05. (B) WB of EndMT markers in WT shCTRL, WT shKLF4, CCM1 KO shCTRL, and CCM1 KO shKLF4 ECs. Vinculin is the loading control. These data are representative of three independent observations. (C) Proliferation rate of WT shCTRL, WTshKLF4, CCM1 KO shCTRL, and CCM1 KO shKLF4 ECs cultured for 5 days. Columns represent mean ± SD (n = 8). A two‐tailed unpaired t‐test was performed. 3 days of culture: ***P = 0.0001, **P = 0.0045, ##P = 0.0017; 5 days of culture: ***P = 0.0001, **P = 0.0029, ##P = 0.0040. (D) Migration rate measured in a wound assay of WT shCTRL, CCM1 KO shCTRL, and CCM1 KO shKLF4 ECs. Mean ± SD is shown (n = 6). A two‐tailed unpaired t‐test was performed. ***P = 0.0004, ###P = 0.0009.ECultured lung‐derived WT ECs were lentivirally transduced with a full‐length murine Klf4 (LentiKLF4) or empty vector (Mock). qRT–PCR (left panel) and WB (right panel) of EndMT markers in Mock and LentiKLF4 ECs. qRT–PCR data are mean ± SD (n = 3) and the fold changes are relative to Mock ECs. A two‐tailed unpaired t‐test was performed. **P = 0.002, ***P = 0.0007, ###P = 0.0001. WB results are representative of three independent observations. Tubulin is the loading control.Source data are available online for this figure. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/26612856), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Human KLF4 by Knockdown Validated

Detection of Human KLF4 by Knockdown Validated

Klf4 significantly rescues the effects of HDAC1 on leukemia cell proliferation. (a) Effects of ectopic expression of Klf4 on HDAC1-induced cell proliferation. * and ** indicate P<0.05 and P<0.01, respectively. (b) FACS analysis showing the effects of ectopic expression of Klf4 on HDAC1-induced cell cycle. * and ** indicate P<0.05 and P<0.01, respectively. (c) CCK8 analysis showing the effects of Klf4 knockdown on HDAC1 deficiency-mediated cell proliferation. * and ** indicate P<0.05 and P<0.01, respectively. (d) FACS analysis showing effects of Klf4 knockdown on HDAC1 deficiency-mediated cell cycle. * and ** indicate P<0.05 and P<0.01, respectively. (e) Western blotting analyses showing the effects of ectopic expression of Klf4 and HDAC1 on the expression levels of p21 and p27. GAPDH was used as the loading control. (f) Western blotting analyses showing the effects Klf4 and HDAC1 knockdown on the expression levels of p21 and p27. GAPDH was used as the loading control. (g) ChIP-PCR assays showing Klf4 binding at the promoter region of p21 and p27 after HDAC1 knockdown. Control or HDAC1-deficient cells were fixed and subjected to ChIP assay with anti-Klf4 antibody. ChIP-PCR primers were used for p21 and p27 promoters where shown in Supplementary Table S2. *, **, and *** indicate P<0.05, P<0.01, and P<0.001, respectively Image collected and cropped by CiteAb from the following publication (https://www.nature.com/articles/cddis2014433), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human KLF4 Antibody

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

5 µg/5 x 106 cells
Sample: BG01V human embryonic stem cell chromatin, B2R promoter detected by standard PCR.

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed BG01V human embryonic stem cells, HT 29 Human Colon Adenocarcinoma Cell Line (positive) and HDLM‑2 Human Hodgkin's Lymphoma Cell Line (negative) Cells

Immunohistochemistry

15 µg/mL
Sample:

Immersion fixed paraffin-embedded sections of human colon

Simple Western

25 µg/mL
Sample: HT‑29 human colon adenocarcinoma cell line

Western Blot

0.5 µg/mL
Sample: HT‑29 human colon adenocarcinoma cell line, SW480 human colorectal adenocarcinoma cell line, and HCT‑116 human colorectal carcinoma cell line

Reviewed Applications

Read 2 reviews rated 4 using AF3640 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

Human KLF4, also known as epithelial zinc finger protein (EZF), is a 53 kDa (470 aa) member of the kruppel C2H2-type zinc finger protein family. It contains three C2H2-type zinc fingers at the carboxyl terminus that preferentially bind to cis-DNA elements that are GC rich. KLF4 regulates the expression of target genes that are involved in different cellular functions. KLF4 is highly expressed in the epithelial cells of the skin and the gastrointestinal tract. Human and mouse KLF4 share 90% amino acid sequence identity.

Long Name

Kruppel-Like Factor 4

Alternate Names

EZF

Entrez Gene IDs

9314 (Human)

Gene Symbol

KLF4

UniProt

AAH29923

Additional KLF4 Products

Product Documents for Human 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 Human KLF4 Antibody

For research use only

Citations for Human KLF4 Antibody

Customer Reviews for Human KLF4 Antibody (2)

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  • Human KLF4 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: A172 human glioblastoma cell line
    Species: Human
    Verified Customer | Posted 01/25/2018
    Human KLF4 Antibody AF3640
  • Name: Anonymous
    Application: Chromatin Immunoprecipitation
    Sample Tested: See PMID 23159369
    Species: Human
    Verified Customer | Posted 01/08/2015

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Protocols

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FAQs for Human KLF4 Antibody

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

    A: AF3640 is against KLF4 and is derived from human. 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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