RecombinantRabMAb

Recombinant Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)

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Flow Cytometry - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
  • Flow Cytometry - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
  • Immunocytochemistry - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)
  • Immunoprecipitation - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
  • Western blot - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
  • Immunoprecipitation - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
  • Western blot - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
  • Western blot - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
  • Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR23501-203] to CD16+CD32
  • Suitable for: Flow Cyt, IP, ICC, WB, IHC-P
  • Reacts with: Mouse, Recombinant fragment

Conjugates logo Related conjugates and formulations

Carrier Free

Overview

  • Product name

    Anti-CD16+CD32 antibody [EPR23501-203]
    See all CD16+CD32 primary antibodies

  • Description

    Rabbit monoclonal [EPR23501-203] to CD16+CD32

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, IP, ICC, WB, IHC-Pmore details
    Unsuitable for: IHC-Fr

  • Species reactivity

    Reacts with: Mouse, Recombinant fragment

  • Immunogen

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

  • Positive control

    • WB: J774A.1 and DC2.4 whole cell lysates; Mouse spleen and liver tissue lysates; His-tagged mouse CD16 recombinant protein; His-tagged mouse CD32 recombinant protein. IHC-P: Mouse spleen, liver and colon carcinoma tissue. ICC: J774A.1 cells. Flow cyt: J774A.1 and mouse splenocyte cells. IP: J774A.1 whole cell lysate; Mouse spleen tissue 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

  • Form

    Liquid

  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.

  • Storage buffer

    Preservative: 0.01% Sodium azide
    Constituents: 59.94% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR23501-203

  • Isotype

    IgG

  • Research areas

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab223200 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
Flow Cyt
1/500.
IP
1/30.
ICC
1/500.
WB
1/1000. Detects a band of approximately 40-60 kDa.
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
Flow Cyt
1/500.
IP
1/30.
ICC
1/500.
WB
1/1000. Detects a band of approximately 40-60 kDa.
IHC-P
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for IHC-Fr.

    • Target

    • Relevance

      Function: Binds to the Fc region of immunoglobulins gamma. Low affinity receptor. By binding to IgG it initiates cellular responses against pathogens and soluble antigens. Promotes phagocytosis of opsonized antigens. Tissue specificity: Found on monocytes, neutrophils and eosinophil platelets. Similarity: Contains 2 Ig-like C2-type (immunoglobulin-like) domains. PTM: Phosphorylated by SRC-type Tyr-kinases such as LYN, BLK, FYN, HCK and SYK.

    • Cellular localization

      CD32: Type I membrane protein. CD16: Attached to the membrane by a GPI anchor. Exists also as a soluble receptor, produced by a proteolytic cleavage.

    • Database links

      • Alternative names

        • CD16a antibody
        • CD16b antibody
        • CD32 antibody
        • CD32A antibody
        • CD32B antibody
        • CDw32 antibody
        • Fc fragment of IgG low affinity IIa receptor antibody
        • Fc fragment of IgG low affinity IIb receptor antibody
        • Fc fragment of IgG low affinity IIIa receptor antibody
        • Fc fragment of IgG low affinity IIIb receptor antibody
        • Fc gamma receptor III A antibody
        • Fc gamma receptor IIIb antibody
        • Fc gamma RIIa antibody
        • Fc gamma RIIb antibody
        • Fc gamma RIII beta antibody
        • Fc-gamma RII-a antibody
        • Fc-gamma-RIIa antibody
        • FCG2A_HUMAN antibody
        • FcGR antibody
        • FCGR2A antibody
        • FCGR2B antibody
        • FCGR3 antibody
        • FCGR3A antibody
        • FCGR3B antibody
        • FcRII-a antibody
        • FcRIII antibody
        • IgG Fc receptor II-a antibody
        • Low affinity immunoglobulin gamma Fc region receptor II-a antibody
        see all

      Images

      • Flow Cytometry - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
        Flow Cytometry - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)

        Flow cytometric analysis of mouse splenocytes cells labelling CD16+CD32 with ab223200 at 1/500 dilution (Right) compared with a Rabbit monoclonal IgG (ab172730) isotype control (Left).

        Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

        Gated on viable cells.

      • Flow Cytometry - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
        Flow Cytometry - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)

        Flow cytometric analysis of bEnd.3 (mouse brain endothelioma, Left) / J774A.1 (mouse reticulum cell sarcoma monocyte macrophage, Right) cells labelling CD16+CD32 with ab223200 at 1/500 dilution (Red) compared with a Rabbit monoclonal IgG (ab172730)  isotype control (Black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue).

        Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) at 1/2000 dilution was used as the secondary antibody.

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

        Gated on viable cells.

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)

        Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD16+CD32 with ab223200 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Cytoplasmic and membranouse staining on mouse spleen (PMID: 22618994). The section was incubated with ab223200 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.

      • Immunocytochemistry - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)
        Immunocytochemistry - Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)

        Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) cells labelling CD16+CD32 with ab223200 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 strong membranous and weak cytoplasmic staining in J774A.1 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.

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

      • Immunoprecipitation - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
        Immunoprecipitation - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)

        CD16+CD32 was immunoprecipitated from 0.35 mg Mouse spleen tissue lysate with ab223200 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab223200 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

        Lane 1: Mouse spleen tissue lysate 10 ug

        Lane 2: ab223200 IP in Mouse spleen tissue lysate

        Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab223200 in mouse spleen tissue lysate

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

        Exposure time: 3 minutes.

         

      • Western blot - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
        Western blot - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
        All lanes : Anti-CD16+CD32 antibody [EPR23501-203] (ab223200) at 1/1000 dilution

        Lane 1 : His-tagged mouse CD16 recombinant protein (aa 31-215), 5 uL
        Lane 2 : His-tagged mouse CD32 recombinant protein (aa 30-210), 25 uL

        Secondary
        All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution


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

        This product can recognize both CD16 and CD32.

        The loading samples are E.coil extracts containing CD16 or CD32 recombinant protein respectively.

        Exposure time: 70 seconds.

         

      • Immunoprecipitation - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
        Immunoprecipitation - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)

        CD16+CD32 was immunoprecipitated from 0.35 mg J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate with ab223200 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab223200 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

        Lane 1: J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) whole cell lysate 10 ug

        Lane 2: ab223200 IP in J774A.1 whole cell lysate

        Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab223200 in J774A.1 whole cell lysate

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

        Exposure time: 8 seconds.

         

      • Western blot - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
        Western blot - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
        All lanes : Anti-CD16+CD32 antibody [EPR23501-203] (ab223200) at 1/1000 dilution

        Lane 1 : Mouse spleen tissue lysate
        Lane 2 : Mouse liver tissue lysate

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

        Observed band size: 40-60 kDa why is the actual band size different from the predicted?



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

        The molecular weight observed is consistent with what has been described in the literature (PMID: 19503602, 31172847).

        Exposure time: 3 minutes.

         

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)

        Immunohistochemical analysis of paraffin-embedded mouse colon carcinoma tissue labeling CD16+CD32 with ab223200 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the stroma in mouse colon carcinoma. The section was incubated with ab223200 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.

      • Western blot - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
        Western blot - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
        All lanes : Anti-CD16+CD32 antibody [EPR23501-203] (ab223200) at 1/1000 dilution

        Lane 1 : J774A.1 (mouse reticum cell sarcoma monocyte macrophage) whole cell lysate
        Lane 2 : bEnd.3 (mouse brain endothelioma) whole cell lysate
        Lane 3 : DC2.4 (mouse dendritic cell) whole cell lysate

        Lysates/proteins at 20 µg per lane.

        Secondary
        All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/50000 dilution

        Observed band size: 40-60 kDa why is the actual band size different from the predicted?



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

        The molecular weight observed is consistent with what has been described in the literature (PMID: 19503602, 31172847).

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

        Exposure time: Lanes 1-2: 5.5 seconds; Lane 3: 59 seconds.

         

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 + CD32 antibody [EPR23501-203] (ab223200)

        Immunohistochemical analysis of paraffin-embedded mouse liver tissue labeling CD16+CD32 with ab223200 at 1/1000 dilution followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse liver. The section was incubated with ab223200 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-CD16+CD32 antibody [EPR23501-203] (ab223200)
        Anti-CD16+CD32 antibody [EPR23501-203] (ab223200)

      Protocols

      To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

      Click here to view the general protocols

      References (0)

      Publishing research using ab223200? Please let us know so that we can cite the reference in this datasheet.

      ab223200 has not yet been referenced specifically in any publications.

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