Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23222-24] to Eph receptor B4/HTK - BSA and Azide free
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free
    See all Eph receptor B4/HTK primary antibodies

  • Description

    Rabbit monoclonal [EPR23222-24] to Eph receptor B4/HTK - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IHC-P, WBmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP

  • Species reactivity

    Reacts with: Mouse, Human

  • Immunogen

    Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK293T, HT-29, HUVEC, HCT116, MCF7, T47D, PC-3, NIH:OVCAR-3, SK-BR-3, GR-M, 4T1 and NIH/3T3 lysates. IHC-P: Human colon, Human colon cancer and Human breast cancer tissues.

  • General notes

    ab269369 is the carrier-free version of ab254301. 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

    This product 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)
    Western blot - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)
    All lanes : Anti-Eph receptor B4/HTK antibody [EPR23222-24] (ab254301) at 1/1000 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : EPHB4 knockout HEK293T cell lysate
    Lane 3 : MCF7 cell lysate
    Lane 4 : HT-29 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 108 kDa
    Observed band size: 125 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab254301).

    Lanes 1-4: Merged signal (red and green). Green - ab254301 observed at 125 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab254301 Anti-Eph receptor B4/HTK antibody [EPR23222-24] was shown to specifically react with Eph receptor B4/HTK in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266733 (knockout cell lysate ab257429) was used. Wild-type and Eph receptor B4/HTK knockout samples were subjected to SDS-PAGE. ab254301 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 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 20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)

    Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling Eph receptor B4 with ab254301 at 1/1000 dilution (0.545ug/ml) followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution. Mainly membranous staining in cancer cells of human breast cancer (PMID: 16816380) is observed. The section was incubated with ab255611 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at Ready to use dilution.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254301).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)

    Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling Eph receptor B4 with ab254301 at 1/1000 dilution (0.545ug/ml) followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution. Positive staining in cancer cells of human colon cancer (PMID: 19366806) is observed. The section was incubated with ab255611 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at Ready to use dilution.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254301).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)

    Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling Eph receptor B4 with ab254301 at 1/1000 dilution (0.545ug/ml) followed by a Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at a ready to use dilution. Weak positive staining in base of crypts of human colon (PMID:19366806) is observed. The section was incubated with ab255611 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at Ready to use dilution.

    Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab254301).

  • Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)
    Anti-Eph receptor B4/HTK antibody [EPR23222-24] - BSA and Azide free (ab269369)

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