Overview

  • Product name

    Anti-KLF4 antibody [EPR20753-25]
    See all KLF4 primary antibodies
  • Description

    Rabbit monoclonal [EPR20753-25] to KLF4
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, ICC/IF, IP, IHC - Wholemountmore details
  • Species reactivity

    Reacts with: Mouse, Rat
  • Immunogen

    Recombinant fragment within Mouse KLF4 aa 1-100. The exact sequence is proprietary.
    Database link: Q60793

  • Positive control

    • WB: F9 and ES-D3 whole cell lysate; Mouse and rat skin and mouse colon tissue lysate. IHC-P: Mouse and rat colon tissue. ICC/IF: F9 cells. IP: F9 whole cell lysate IHC-Wm: E3.75 mouse blastocyst
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Applications

Our Abpromise guarantee covers the use of ab214666 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
IHC-P 1/1000.
WB 1/1000. Predicted molecular weight: 54 kDa.
ICC/IF 1/1000.
IP 1/30.
IHC - Wholemount 1/1000.

Target

  • 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
  • Database links

  • Alternative names

    • Endothelial Kruppel like zinc finger protein antibody
    • Epithelial zinc finger protein EZF antibody
    • EZF antibody
    • GKLF antibody
    • gut Kruppel-like factor antibody
    • Gut-enriched krueppel-like factor antibody
    • KLF antibody
    • KLF4 antibody
    • KLF4_HUMAN antibody
    • Krueppel-like factor 4 antibody
    • Kruppel like factor 4 (Epithelial zinc finger protein EZF) (Gut enriched Krueppel like factor) antibody
    • Kruppel like factor 4 (gut) antibody
    see all

Images

  • Several embryos at the mid-blastocyst stage (~E3.75) were tested. The embryos were fixed for 10 mins at room temperature in 4% PFA. Fixed embryos were washed for 5 mins in PBS + 0.1% Triton X-100 (PBX) at room temperature. The embryos were then permeabilized in PBS + 0.5% Triton X-100 + 100mM Glycine for 5 mins at room temperature and washed again in PBX for 5 mins at room temperature. Blocking was carried out for 1 hour at room temperature in PBS + 2% horse serum and then the embryos were incubated overnight in ab214666 (1/1000) diluted in blocking solution. The next day embryos were washed three times for 5 mins each in PBX, before blocking again for 1 hour at room temperature. Then incubated in Donkey anti rabbit Alexa Fluor® 488 secondary antibodies (1/500) diluted in blocking solution for 1 hour at 4ºC. This was followed by 2 washes of 5 mins each in PBX. The embryos were then incubated in PBS + 5μg/ml DAPI for 10 min, or until mounting for imaging. Images were acquired on a laser-scanning confocal microscope system. Displayed image shows maximum intensity projection of 5 optical sections acquired at 1μm increments. Scale bar: 20 μm

  • Lanes 1-4 : Anti-KLF4 antibody [EPR20753-25] (ab214666) at 1/1000 dilution
    Lane 5 : Anti-KLF4 antibody [EPR20753-25] (ab214666) at 1/5000 dilution

    Lane 1 : F9 (mouse embryonal carcinoma epithelial cell), whole cell lysate
    Lane 2 : Mouse skin tissue lysate
    Lane 3 : Mouse colon tissue lysate
    Lane 4 : Rat skin tissue lysate
    Lane 5 : ES D3 (mouse embryonic stem cell), whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Lanes 1-4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
    Lane 5 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 54 kDa
    Observed band size: 52 kDa
    why is the actual band size different from the predicted?



    Exposure times:
    Lanes 1-2: 3 minutes
    Lane 3-4: 92 seconds
    Lane 5: 48 seconds

    Blocking/Diluting buffer and concentration: 5% NFDM/TBST

     

  • KLF4 was immunoprecipitated from 0.35 mg F9 (Mouse embryonal carcinoma epithelial cell) whole cell lysate using ab214666 at 1/30 dilution. Western blot was performed on the immunoprecipitate using ab214666 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used as the secondary at 1/5000 dilution.

    Lane 1: F9 whole cell lysate 10 µg (input).
    Lane 2: ab214666 IP in F9 whole cell lysate.
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214666 in F9 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 3 minutes.

    Fresh lysate was required and IP test was conducted immediately. Incubation time was shortened from overnight to 2h.

  • Immunohistochemical analysis of paraffin-embedded Rat colon tissue labeling KLF4 with ab214666 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody. Nuclear staining on epithelial cells of rat colon (PMID: 21070761, PMID: 19109406) is observed. Counterstained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

  • Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling KLF4 with ab214666 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP) secondary antibody. Nuclear staining on epithelial cells of mouse colon (PMID: 21070761, PMID: 19109406) is observed. Counterstained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized F9 (mouse embryonal carcinoma epithelial cell) cells labeling KLF4 with ab214666 at 1/100 dilution, followed by a AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000 dilution (Green). Confocal image showing nuclear staining in F9 cell line is observed. The nuclear counterstain was DAPI (Blue). Tubulin was counterstained using an Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594, ab195889, Red) at 1.200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a AlexaFluor®488 Goat anti-Rabbit secondary (ab150077) at 1/1000.

References

ab214666 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Application
IHC - Wholemount
Sample
Mouse Embryo (E3.75 mouse blastocyst)
Specification
E3.75 mouse blastocyst

Mr. Vidur Garg

Verified customer

Submitted Jul 08 2019

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