Human KLF4 Antibody

(8 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human KLF4 in direct ELISAs and Western blots. In direct ELISAs, this antibody shows less than 30% cross‑reactivity with recombinant mouse KLF4.
  • Source
    Polyclonal Goat IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human KLF4
    Ala2-Phe470
    Accession # AAH29923
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    0.5 µg/mL
    See below
  • Simple Western
    25 µg/mL
    See below
  • Immunohistochemistry
    15 µg/mL
    See below
  • Chromatin Immunoprecipitation (ChIP)
    5 µg/5 x 106 cells
    See below
  • Immunocytochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
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 (Catalog # 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.
Chromatin Immunoprecipitation (ChIP)
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 (Catalog # AB-108-C) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Goat IgG Secondary Antibody (Catalog # BAF109). Immunocomplexes were captured using 50 μL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. The B2R promoter was detected by standard PCR.
Immunocytochemistry
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; Catalog # NL001) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
Immunohistochemistry
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 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 (Catalog # 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.

Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • 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.
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
  • Entrez Gene IDs:
    9314 (Human)
  • Alternate Names:
    Epithelial zinc finger protein EZF; EZF; EZFendothelial Kruppel-like zinc finger protein; GKLFKrueppel-like factor 4; Gut-enriched krueppel-like factor; KLF4; Kruppel-like factor 4 (gut)
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

8 Citations: Showing 1 - 8
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Species
Applications
Sample Type
  1. HIF1? regulates single differentiated glioma cell dedifferentiation to stem-like cell phenotypes with high tumorigenic potential under hypoxia
    Authors: P Wang, C Lan, S Xiong, X Zhao, Y Shan, R Hu, W Wan, S Yu, B Liao, G Li, J Wang, D Zou, B Chen, H Feng, N Wu
    Oncotarget, 2017;8(17):28074-28092.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  2. KLF2 and KLF4 control endothelial identity and vascular integrity
    Authors: P Sangwung, G Zhou, L Nayak, ER Chan, S Kumar, DW Kang, R Zhang, X Liao, Y Lu, K Sugi, H Fujioka, H Shi, SD Lapping, CC Ghosh, SJ Higgins, SM Parikh, H Jo, MK Jain
    JCI Insight, 2017;2(4):e91700.
    Species: Human
    Sample Type: Cell Lysates
    Application: WB
  3. Induced overexpression of Oct4A in human dental pulp cells enhances pluripotency and multilineage differentiation capability.
    Authors: Liu L, Wu L, Wei X, Ling J
    Stem Cells Dev, 2015;24(8):962-72.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC Fluorescence
  4. Lymphatic regulator PROX1 determines Schlemm's canal integrity and identity.
    Authors: Park D, Lee J, Park I, Choi D, Lee S, Song S, Hwang Y, Hong K, Nakaoka Y, Makinen T, Kim P, Alitalo K, Hong Y, Koh G
    J Clin Invest, 2014;124(9):3960-74.
  5. ZNF750 interacts with KLF4 and RCOR1, KDM1A, and CTBP1/2 chromatin regulators to repress epidermal progenitor genes and induce differentiation genes.
    Authors: Boxer L, Barajas B, Tao S, Zhang J, Khavari P
    Genes Dev, 2014;28(18):2013-26.
    Species: Human
    Sample Type: Cell Lysates
    Application: IP
  6. KLF4 modulates expression of IL-6 in dendritic cells via both promoter activation and epigenetic modification.
    Authors: Rosenzweig, Jason M, Glenn, Justin D, Calabresi, Peter A, Whartenby, Katharin
    J Biol Chem, 2013;288(33):23868-74.
    Species: Human
    Sample Type: Cell Lysates
    Application: EMSA
  7. Endogenous Purification Reveals GREB1 as a Key Estrogen Receptor Regulatory Factor.
    Authors: Mohammed H
    Oncogene, 2012;3(2):342-9.
    Species: Human
    Sample Type: Cell Lysates
    Application: ChIP
  8. Innate immune suppression enables frequent transfection with RNA encoding reprogramming proteins.
    Authors: Angel M, Yanik MF,
    PLoS ONE, 2010;5(7):e11756.
    Species: Human
    Sample Type: Whole Cells
    Application: ICC
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