Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CD40 antibody [EPR23890-151] (ab252428)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23890-151] to CD40
  • Suitable for: WB, IHC-P, IP
  • Reacts with: Mouse

Overview

  • Product name

    Anti-CD40 antibody [EPR23890-151]
    See all CD40 primary antibodies

  • Description

    Rabbit monoclonal [EPR23890-151] to CD40

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    IHC-P
    Mouse
    IP
    Mouse
    WB
    Mouse
    See all applications and species data

  • Immunogen

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

  • Positive control

    • WB: RAW 264.7 treated with/ml LPS, Mouse lymph node lysates. IHC-P: Mouse spleen, Mouse colon tissues. IP: RAW 264.7 cell lysate

  • General notes

    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-CD40 antibody [EPR23890-151] (ab252428)
    Western blot - Anti-CD40 antibody [EPR23890-151] (ab252428)
    All lanes : Anti-CD40 antibody [EPR23890-151] (ab252428) at 1/1000 dilution

    Lane 1 : Untreated RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) whole cell lysate
    Lane 2 : RAW 264.7 treated with 1 µg/ml LPS (Lipopolysaccharide) for 6 hours whole cell lysate
    Lane 3 : Mouse lymph node tissue lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Lane 1 : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution
    Lanes 2-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

    Predicted band size: 30 kDa
    Observed band size: 40 kDa
    why is the actual band size different from the predicted?



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

    LPS induces the expression of CD40 in Raw 264.7 cells (PMID: 23605514) 

    Exposure time: Lanes 1-2: 10 seconds Lane 3: 48 seconds.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR23890-151] (ab252428)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR23890-151] (ab252428)

    Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CD40 with ab252428 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Membranous and cytoplasmic staining on mouse spleen (PMID: 25316820) is observed. The section was incubated with ab252428 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

  • Immunoprecipitation - Anti-CD40 antibody [EPR23890-151] (ab252428)
    Immunoprecipitation - Anti-CD40 antibody [EPR23890-151] (ab252428)

    CD40 was immunoprecipitated from 0.35 mg RAW 264.7 (mouse abelson murine leukemia virus-induced tumor macrophage) treated with 1 ug/ml LPS (Lipopolysaccharide) for 6 hours whole cell lysate with ab252428 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab252428 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

    Lane 1: RAW 264.7 treated with 1 ug/ml LPS (Lipopolysaccharide) for 6 hours whole cell lysate 10 ug

    Lane 2: ab252428 IP in RAW 264.7 whole cell lysate

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab252428 in RAW 264.7 whole cell lysate

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

    Exposure time: 3 minutes

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR23890-151] (ab252428)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD40 antibody [EPR23890-151] (ab252428)

    Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling CD40 with ab252428 at 1/500 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on stromal cells of mouse colon is observed. The section was incubated with ab252428 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 Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

  • Anti-CD40 antibody [EPR23890-151] (ab252428)
    Anti-CD40 antibody [EPR23890-151] (ab252428)

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