Recombinant
RabMAb

Recombinant Anti-CD8B antibody [EPR22331-54] (ab228965)

Overview

  • Product name

    Anti-CD8B antibody [EPR22331-54]
    See all CD8B primary antibodies
  • Description

    Rabbit monoclonal [EPR22331-54] to CD8B
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, IHC-P, WB, IHC-Frmore details
    Unsuitable for: Flow Cyt
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

    Recombinant fragment within Mouse CD8B aa 1-200. The exact sequence is proprietary.
    Database link: P10300

  • Positive control

    • WB: Mouse spleen, lymph node and thymus lysates. IP: Mouse thymus lysate. IHC-P: Mouse spleen and thymus tissue. IHC-Fr: Mouse thymus 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

  • 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: PBS, 0.05% BSA, 40% Glycerol
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR22331-54
  • Isotype

    IgG

Applications

Our Abpromise guarantee covers the use of ab228965 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
IP 1/30.
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Predicted molecular weight: 24 kDa.
IHC-Fr 1/100.
  • Application notes
    Is unsuitable for Flow Cyt.
  • Target

    • Cellular localization

      Isoform 1: Cell membrane; Single-pass type I membrane protein (Probable). Isoform 2: Cell membrane; Single-pass type I membrane protein (Probable). Isoform 3: Secreted (Probable). Isoform 4: Cell membrane; Single-pass type I membrane protein (Probable). Isoform 5: Cell membrane; Single-pass type I membrane protein (Probable). Isoform 6: Secreted (Probable). Isoform 7: Secreted (Probable). Isoform 8: Secreted (Probable).
    • Database links

    • Alternative names

      • CD_antigen=CD8b antibody
      • CD8 antigen 37 kDa chain antibody
      • CD8 antigen, beta chain antibody
      • CD8 antigen, beta polypeptide 1 (p37) antibody
      • CD8 antigen, beta polypeptide 1 antibody
      • CD8B antibody
      • CD8b molecule antibody
      • CD8B1 antibody
      • Flags: Precursor antibody
      • LEU2 antibody
      • LY3 antibody
      • LYT3 antibody
      • MGC119115 antibody
      • OTTHUMP00000160762 antibody
      • OTTHUMP00000160763 antibody
      • OX 8 membrane antigen antibody
      • P37 antibody
      • T cell surface glycoprotein CD8 antibody
      • T cell surface glycoprotein CD8 beta chain antibody
      • T lymphocyte surface glycoprotein beta chain antibody
      see all

    Images

    • CD8B was immunoprecipitated from 0.35 mg of mouse thymus lysate with ab228965 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab228965 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.

      Lane 1: Mouse thymus lysate 10 μg (Input).
      Lane 2: ab228965 IP in mouse thymus lysate.
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab22896 in mouse thymus lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.
      Exposure time: 3 seconds.

    • Frozen section of 4% PFA-fixed, 0.2% Triton X-100 permeabilized mouse thymus tissue stained for CD8B (green) using ab228965 at 1/100 dilution in immunohistochemical analysis. The secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution. Specific membrane staining on T cells in the cortex of mouse thymus (PMID 12702501) is observed. The nuclear conter stain is DAPI (blue).

      Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

      Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

       

    • Immunohistochemical analysis of paraffin-embedded mouse thymus tissue labeling CD8B with ab228965 at 1/500 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on T cells in the cortex of mouse thymus (PMID: 12702501) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

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

      The section was incubated with ab228965 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

       

    • Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD8B with ab228965 at 1/500 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on T cells in periarterial lymphatic sheath of mouse spleen (PMID: 18776164) is observed. Counter stained with hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, the ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody.

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

      The section was incubated with ab228965 for 30 mins at RT. The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    • All lanes : Anti-CD8B antibody [EPR22331-54] (ab228965) at 1/1000 dilution

      Lane 1 : Mouse spleen lysate
      Lane 2 : Mouse lymph node lysate
      Lane 3 : Mouse thymus lysate

      Lysates/proteins at 10 µg per lane.

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

      Predicted band size: 24 kDa
      Observed band size: 28-32 kDa why is the actual band size different from the predicted?



      Blocking and dilution buffer: 5% NFDM/TBST.

      CD8B is a glycoprotein. The molecular weight observed is consistent with what has been described in the literature (PMID: 11719190).

      Exposure times.

      Lanes 1-2: 37 seconds. Lane 3: 6 seconds.

    References

    ab228965 has not yet been referenced specifically in any publications.

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