Overview

  • Product name

  • Description

    Rabbit polyclonal to KLF4
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse
    Predicted to work with: Rat, Chinese hamster
  • Immunogen

    Synthetic peptide corresponding to Mouse KLF4 aa 1-100 conjugated to Keyhole Limpet Haemocyanin (KLH).
    Database link: Q60793

  • Positive control

    • WB: Mouse ES-E14TG2a whole cell lysate. ICC/IF: mES cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab129473 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
ICC/IF Use a concentration of 1 µg/ml.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 60 kDa (predicted molecular weight: 54 kDa).

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

  • Anti-KLF4 antibody (ab129473) at 1 µg/ml + E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 54 kDa
    Observed band size: 60 kDa
    why is the actual band size different from the predicted?


    Exposure time: 8 minutes


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab129473 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

  • ICC/IF image of ab129473 stained mouse ES cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab129473, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM. This antibody also gave a positive result in 100% methanol fixed (5 min) mouse ES cells at 1µg/ml.

References

This product has been referenced in:

  • Mathieu J  et al. Folliculin regulates mTORC1/2 and WNT pathways in early human pluripotency. Nat Commun 10:632 (2019). Read more (PubMed: 30733432) »
  • Kaowinn S  et al. CGK062, a small chemical molecule, inhibits cancer upregulated gene 2-induced oncogenesis through NEK2 and ß-catenin. Int J Oncol 54:1295-1305 (2019). Read more (PubMed: 30968157) »
See all 7 Publications for this product

Customer reviews and Q&As

1-4 of 4 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (primary astrocytes)
Permeabilization
No
Specification
primary astrocytes
Blocking step
Serum as blocking agent for 45 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Acetone

Abcam user community

Verified customer

Submitted Sep 10 2018

Application
Western blot
Loading amount
15 µg
Gel Running Conditions
Reduced Denaturing
Sample
Mouse Cell lysate - whole cell (mouse embryonic stem cells (46C))
Specification
mouse embryonic stem cells (46C)
Treatment
transfection with Klf4 expression vector (Flag-tagged Klf4)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

Abcam user community

Verified customer

Submitted Apr 04 2014

Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 0.1% · Temperature: 21°C
Sample
Mouse Cell (D3 Mouse Embryonic Stem Cell line)
Specification
D3 Mouse Embryonic Stem Cell line
Permeabilization
Yes - 0.1% Triton X-100
Fixative
Paraformaldehyde

Joe Segal

Verified customer

Submitted Jul 18 2013

Application
Western blot
Loading amount
20000 cells
Gel Running Conditions
Reduced Denaturing (gel 10%)
Sample
Mouse Cell lysate - whole cell (D3 Mouse Embryonic Stem Cell line)
Specification
D3 Mouse Embryonic Stem Cell line
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C

Joe Segal

Verified customer

Submitted Jul 02 2013

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