Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-KLF4 antibody [EPR20753-25] - BSA and Azide free (ab255335)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR20753-25] to KLF4 - BSA and Azide free
  • Suitable for: IHC-P, ICC/IF, IP, IHC - Wholemount, WB
  • Reacts with: Mouse, Rat

Overview

  • Product name

    Anti-KLF4 antibody [EPR20753-25] - BSA and Azide free
    See all KLF4 primary antibodies

  • Description

    Rabbit monoclonal [EPR20753-25] to KLF4 - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, ICC/IF, IP, IHC - Wholemount, WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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.

  • General notes

    ab255335 is the carrier-free version of ab214666. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab255335 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

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

Images

  • IHC - Wholemount - Anti-KLF4 antibody [EPR20753-25] - BSA and Azide free (ab255335)
    IHC - Wholemount - Anti-KLF4 antibody [EPR20753-25] - BSA and Azide free (ab255335) Image courtesy of Vidur Garg from the Sloan-Kettering Institute

    This data was developed using ab214666, the same antibody clone in a different buffer formulation.

    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

  • Immunoprecipitation - Anti-KLF4 antibody [EPR20753-25] - BSA and Azide free (ab255335)
    Immunoprecipitation - Anti-KLF4 antibody [EPR20753-25] - BSA and Azide free (ab255335)

    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214666).
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KLF4 antibody [EPR20753-25] - BSA and Azide free (ab255335)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KLF4 antibody [EPR20753-25] - BSA and Azide free (ab255335)

    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214666).

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KLF4 antibody [EPR20753-25] - BSA and Azide free (ab255335)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KLF4 antibody [EPR20753-25] - BSA and Azide free (ab255335)

    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).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214666).

    Perform heat mediated antigen retrieval with EDTA buffer pH 9 before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-KLF4 antibody [EPR20753-25] - BSA and Azide free (ab255335)
    Immunocytochemistry/ Immunofluorescence - Anti-KLF4 antibody [EPR20753-25] - BSA and Azide free (ab255335)

    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).


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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214666).
  • Anti-KLF4 antibody [EPR20753-25] - BSA and Azide free (ab255335)
    Anti-KLF4 antibody [EPR20753-25] - BSA and Azide free (ab255335)

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