Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-CD16+CD32 antibody [EPR23501-203] - BSA and Azide free (ab275095)

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 - BSA and Azide free
  • Suitable for: WB, IP, IHC-P, ICC, Flow Cyt
  • Reacts with: Mouse

Overview

  • Product name

    Anti-CD16+CD32 antibody [EPR23501-203] - BSA and Azide free
    See all CD16+CD32 primary antibodies

  • Description

    Rabbit monoclonal [EPR23501-203] to CD16+CD32 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Mouse
    ICC
    Mouse
    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: 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

    ab275095 is the carrier-free version of ab223200. 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

    ab275095 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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Flow Cytometry - Anti-CD16 + CD32 antibody [EPR23501-203] - BSA and Azide free (ab275095)
    Flow Cytometry - Anti-CD16 + CD32 antibody [EPR23501-203] - BSA and Azide free (ab275095)

    This data was developed using ab223200, the same antibody clone in a different buffer formulation.

    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] - BSA and Azide free (ab275095)
    Flow Cytometry - Anti-CD16 + CD32 antibody [EPR23501-203] - BSA and Azide free (ab275095)

    This data was developed using ab223200, the same antibody clone in a different buffer formulation.

    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] - BSA and Azide free (ab275095)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 + CD32 antibody [EPR23501-203] - BSA and Azide free (ab275095)

    This data was developed using ab223200, the same antibody clone in a different buffer formulation.

    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] - BSA and Azide free (ab275095)
    Immunocytochemistry - Anti-CD16+CD32 antibody [EPR23501-203] - BSA and Azide free (ab275095)

    This data was developed using ab223200, the same antibody clone in a different buffer formulation.

    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] - BSA and Azide free (ab275095)
    Immunoprecipitation - Anti-CD16 + CD32 antibody [EPR23501-203] - BSA and Azide free (ab275095)

    This data was developed using ab223200, the same antibody clone in a different buffer formulation.

    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] - BSA and Azide free (ab275095)
    Western blot - Anti-CD16 + CD32 antibody [EPR23501-203] - BSA and Azide free (ab275095)
    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


    This data was developed using ab223200, the same antibody clone in a different buffer formulation. 

    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] - BSA and Azide free (ab275095)
    Immunoprecipitation - Anti-CD16 + CD32 antibody [EPR23501-203] - BSA and Azide free (ab275095)

    This data was developed using ab223200, the same antibody clone in a different buffer formulation.

    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] - BSA and Azide free (ab275095)
    Western blot - Anti-CD16 + CD32 antibody [EPR23501-203] - BSA and Azide free (ab275095)
    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?



    This data was developed using ab223200, the same antibody clone in a different buffer formulation. 

    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] - BSA and Azide free (ab275095)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 + CD32 antibody [EPR23501-203] - BSA and Azide free (ab275095)

    This data was developed using ab223200, the same antibody clone in a different buffer formulation.

    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] - BSA and Azide free (ab275095)
    Western blot - Anti-CD16 + CD32 antibody [EPR23501-203] - BSA and Azide free (ab275095)
    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?



    This data was developed using ab223200, the same antibody clone in a different buffer formulation. 

    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] - BSA and Azide free (ab275095)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 + CD32 antibody [EPR23501-203] - BSA and Azide free (ab275095)

    This data was developed using ab223200, the same antibody clone in a different buffer formulation.

    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] - BSA and Azide free (ab275095)
    Anti-CD16+CD32 antibody [EPR23501-203] - BSA and Azide free (ab275095)

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