Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR22714-2] to FOXP4 - BSA and Azide free
  • Suitable for: IHC-Fr, WB, Flow Cyt (Intra), IP, IHC-P, ICC
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free
    See all FOXP4 primary antibodies

  • Description

    Rabbit monoclonal [EPR22714-2] to FOXP4 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt (Intra)
    Human
    ICC
    Human
    IHC-Fr
    Mouse
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: HepG2, Caco-2, HeLa, Hepa1-6, 4T1 and PC-12 whole cell lysates; Human colon and tonsil tissue lysates; Rat spleen tissue lysate. IHC-P: Human tonsil tissue; Mouse spleen tissue; Rat spleen tissue. IHC-Fr: Mouse colon tissue. ICC: HepG2 cells. Flow cyt: HepG2 cells. IP: HepG2 whole cell lysate.

  • General notes

    ab279342 is the carrier-free version of ab242127. 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

    ab279342 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.

Properties

Images

  • Western blot - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)
    Western blot - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)
    All lanes : Anti-FOXP4 antibody [EPR22714-2] (ab242127) at 1/1000 dilution

    Lane 1 : HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate
    Lane 2 : FOXP4 knock out HepG2 whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (IRDye® 800CW) (ab216773) and Goat Anti-Mouse IgG H&L (IRDye® 680RD) (ab216776), 1/10000 dilution at 1/10000 dilution

    Predicted band size: 73 kDa
    Observed band size: 80 kDa
    why is the actual band size different from the predicted?



    This data was developed using ab242127, the same antibody clone in a different buffer formulation. 

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

    The lysates were kindly provided by our collaborator Dr. Bin Zhao, Zhejiang University.

     

    Lanes 1-2: Merged signal (red and green). Green - ab242127 observed at 80 kDa. Red - loading control ab8245 (Mouse monoclonal [6C5] to GAPDH) observed at 36 kDa.

     

    Lanes 1-2: ab242127 Anti-FOXP4 antibody [EPR22714-2] was shown to react with FOXP4 in HepG2 cells in Western blot. Loss of signal was observed when FOXP4 knockout sample was used. Wild-type and FOXP4 knockout samples were subjected to SDS-PAGE. ab242127 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at 4°C overnight at 1 in 1000 dilution and 1 in 10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 10000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)
    Western blot - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)
    All lanes : Anti-FOXP4 antibody [EPR22714-2] (ab242127) at 1/1000 dilution

    Lane 1 : HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate
    Lane 2 : Caco-2 (human colorectal adenocarcinoma epithelial cell) whole cell lysate
    Lane 3 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate
    Lane 4 : Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 73 kDa
    Observed band size: 80 kDa why is the actual band size different from the predicted?



    This data was developed using ab242127, the same antibody clone in a different buffer formulation. 

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

    Lysates were made freshly and used in WB immediately to minimize protein degradation.

    Exposure time: Lanes 1-2: 59 seconds; Lanes 3-4: 3 minutes.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)

    This data was developed using ab242127, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Human tonsil tissue labeling FOXP4 with ab242127 at 1/4000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on human tonsil. The section was incubated with ab242127 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

  • Immunocytochemistry - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)
    Immunocytochemistry - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)

    This data was developed using ab242127, the same antibody clone in a different buffer formulation.

    Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HepG2 cells labelling FOXP4 with ab242127 at 1/500 dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing nuclear staining in HepG2 cells. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

    Secondary antibody only control: Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000  dilution.

  • Flow Cytometry (Intracellular) - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)
    Flow Cytometry (Intracellular) - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)

    This data was developed using ab242127, the same antibody clone in a different buffer formulation.

    Flow cytometric analysis of 4% paraformaldehyde fixed, 90% methanol permeabilized HepG2 (human hepatocellular carcinoma epithelial cell) cells labelling FOXP4 with ab242127 at 1/50 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

  • Immunoprecipitation - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)
    Immunoprecipitation - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)

    This data was developed using ab242127, the same antibody clone in a different buffer formulation.

    FOXP4 was immunoprecipitated from 0.35 mg HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate with ab242127 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab242127 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate 10 ug

    Lane 2: ab242127 IP in HepG2 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab242127 in HepG2 whole cell lysate

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

    Exposure time: 8 seconds

     

  • Western blot - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)
    Western blot - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)
    All lanes : Anti-FOXP4 antibody [EPR22714-2] (ab242127) at 1/1000 dilution

    Lane 1 : Human colon tissue lysate
    Lane 2 : Human tonsil tissue lysate
    Lane 3 : Rat spleen tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Lanes 1-2 : VeriBlot for IP secondary antibody(HRP)(ab131366) at 1/1000 dilution
    Lane 3 : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 dilution

    Predicted band size: 73 kDa
    Observed band size: 80 kDa why is the actual band size different from the predicted?



    This data was developed using ab242127, the same antibody clone in a different buffer formulation. 

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

    Bands around 50 kDa and 37 kDa in lane3 are caused by degradation.

    Exposure time: 3 minutes

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)

    This data was developed using ab242127, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling FOXP4 with ab242127 at 1/4000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on mouse spleen. The section was incubated with ab242127 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

  • Immunohistochemistry (Frozen sections) - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)
    Immunohistochemistry (Frozen sections) - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)

    This data was developed using ab242127, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse colon tissue labeling FOXP4 with ab242127 at 1/1000 dilution followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution (Green). Nuclear staining on mouse colon is observed. The nuclear counterstain was DAPI (Blue).

    Secondary antibody control: Secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at 1/1000 dilution.

    Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

  • Western blot - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)
    Western blot - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)
    All lanes : Anti-FOXP4 antibody [EPR22714-2] (ab242127) at 1/1000 dilution

    Lane 1 : 4T1 (mouse mammary gland carcinoma epithelial cell) whole cell lysate
    Lane 2 : PC-12 (rat adrenal gland pheochromocytoma ) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)

    Predicted band size: 73 kDa
    Observed band size: 80 kDa why is the actual band size different from the predicted?



    This data was developed using ab242127, the same antibody clone in a different buffer formulation. 

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

    Lysates were made freshly and used in WB immediately to minimize protein degradation.

    Exposure time: 3 minutes

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)

    This data was developed using ab242127, the same antibody clone in a different buffer formulation.

    Immunohistochemical analysis of paraffin-embedded Rat spleen tissue labeling FOXP4 with ab242127 at 1/4000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Nuclear staining on rat spleen. The section was incubated with ab242127 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

    Secondary antibody only control: Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

    Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

  • Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)
    Anti-FOXP4 antibody [EPR22714-2] - BSA and Azide free (ab279342)

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