Recombinant
RabMAb

Recombinant Anti-CD16 + CD32 antibody [EPR22345] (ab228971)

Overview

  • Product name

    Anti-CD16 + CD32 antibody [EPR22345]
  • Description

    Rabbit monoclonal [EPR22345] to CD16 + CD32
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Pmore details
    Unsuitable for: Flow Cyt,ICC/IF or IP
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

    Recombinant fragment within Mouse CD16 aa 1-200. The exact sequence is proprietary.
    Database link: P08508

  • Positive control

    • WB: Mouse spleen, liver and thymus lysate. His-tagged mouse CD16 recombinant protein(aa31-215) and His-tagged mouse CD32 recombinant protein (aa30-210). J774A.1 and DC2.4 whole cell lysate. IHC-P: Mouse liver and colon tissue.
  • General notes

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab228971 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
WB 1/1000. Predicted molecular weight: 29 kDa.
IHC-P 1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for Flow Cyt,ICC/IF or IP.
  • Target

    Images

    • Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling CD16 with ab228971 at 1/4,000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on Kupffer cells and sinusoidal cells of mouse liver (PMID:26512139) 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).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • All lanes : Anti-CD16 + CD32 antibody [EPR22345] (ab228971) at 1/1000 dilution

      Lane 1 : Mouse spleen lysate
      Lane 2 : Mouse thymus lysate
      Lane 3 : Mouse liver lysate
      Lane 4 : J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate
      Lane 5 : DC2.4 (mouse dendritic cell) whole cell lysate
      Lane 6 : bEnd.3 (mouse brain endothelioma) 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

      Predicted band size: 29 kDa
      Observed band size: 30-55 kDa
      why is the actual band size different from the predicted?



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure times: Lanes 1,2,3 and 6: 3 minutes; Lanes 4 and 5: 15 seconds.

      Bands between 30-55 KD are likely to be glycosylated forms of CD16 and CD32 (PMID: 9378972, PMID: 19503602).

      Negative control: bEnd.3 cell line (PMID: 23911392).

       

       

       

    • All lanes : Anti-CD16 + CD32 antibody [EPR22345] (ab228971) at 1/1000 dilution

      Lane 1 : His-tagged mouse CD16 recombinant protein (aa31-215), 20 ng
      Lane 2 : His-tagged mouse CD32 recombinant protein (aa30-210), 20 ng

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

      Predicted band size: 29 kDa
      Observed band size: 21 kDa why is the actual band size different from the predicted?


      Exposure time: 6 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling CD16 with ab228971 at 1/4,000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Cytoplasmic staining on stromal cells in mouse colon (PMID:26512139) 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).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    References

    ab228971 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

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    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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