Recombinant
RabMAb

Recombinant Anti-CD4 antibody [EPR19514] - Low endotoxin, Azide free (ab221775)

Overview

  • Product name

    Anti-CD4 antibody [EPR19514] - Low endotoxin, Azide free
    See all CD4 primary antibodies
  • Description

    Rabbit monoclonal [EPR19514] to CD4 - Low endotoxin, Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, IHC-Fr, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse
  • Immunogen

    Recombinant fragment aa 1-400. The exact sequence is proprietary.
    Database link: P06332

  • Positive control

    • WB: Mouse thymus lysate; EL4 whole cell lysate. IHC-P: Mouse spleen and colon tissues. IHC-Fr: Mouse spleen tissue. IP: Mouse thymus whole cell lysate.
  • General notes

    ab221775 is a carrier-free antibody designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our Low endotoxin, azide-free formats have low endotoxin level (≤ 1 EU/ml, determined by the LAL assay) and are free from azide, to achieve consistent experimental results in functional assays.

    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 ab221775 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 Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.

Antigen retrieval: Heated citrate solution (10mM citrate PH 6.0 + 0.05% Tween-20).

WB Use at an assay dependent concentration. Detects a band of approximately 51 kDa (predicted molecular weight: 51 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Target

  • Function

    Accessory protein for MHC class-II antigen/T-cell receptor interaction. May regulate T-cell activation. Induces the aggregation of lipid rafts.
  • Sequence similarities

    Contains 3 Ig-like C2-type (immunoglobulin-like) domains.
    Contains 1 Ig-like V-type (immunoglobulin-like) domain.
  • Post-translational
    modifications

    Palmitoylation and association with LCK contribute to the enrichment of CD4 in lipid rafts.
  • Cellular localization

    Cell membrane. Localizes to lipid rafts. Removed from plasma membrane by HIV-1 Nef protein that increases clathrin-dependent endocytosis of this antigen to target it to lysosomal degradation. Cell surface expression is also down-modulated by HIV-1 Envelope polyprotein gp160 that interacts with, and sequesters CD4 in the endoplasmic reticulum.
  • Information by UniProt
  • Database links

  • Alternative names

    • CD 4 antibody
    • CD4 (L3T4) antibody
    • CD4 antibody
    • CD4 antigen (p55) antibody
    • CD4 antigen antibody
    • CD4 molecule antibody
    • CD4 receptor antibody
    • CD4+ Lymphocyte deficiency, included antibody
    • CD4_HUMAN antibody
    • CD4mut antibody
    • L3T4 antibody
    • Leu3 antibody
    • Ly-4 antibody
    • Lymphocyte antigen CD4 antibody
    • MGC165891 antibody
    • OTTHUMP00000238897 antibody
    • p55 antibody
    • T cell antigen T4 antibody
    • T cell antigen T4/LEU3 antibody
    • T cell differentiation antigen L3T4 antibody
    • T cell OKT4 deficiency, included antibody
    • T cell surface antigen T4/Leu 3 antibody
    • T cell surface antigen T4/Leu3 antibody
    • T cell surface glycoprotein CD4 antibody
    • T-cell surface antigen T4/Leu-3 antibody
    • T-cell surface glycoprotein CD4 antibody
    • W3/25 antibody
    • W3/25 antigen antibody
    see all

Images

  • 10% NBF-fixed, paraffin-embedded mouse spleen tissue stained for CD4 using ab183685 at 1/2000 dilution in immunohistochemical analysis, followed by Goat anti-Rabbit IgG Alexa Fluor® 647.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183685).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Formaldehyde-fixed, paraffin-embedded mouse thymus tissue stained for CD4 using ab183685 at 1/500 dilution in immunohistochemical analysis.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183685).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling CD4 with ab183685 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
    Membrane staining on lymphocytes and negative on epithelium cells of mouse colon is observed.
    Counter stained with Hematoxylin.
     
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183685).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling CD4 with ab183685 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

    Negative on mouse cerebrum.

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183685).

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen Mouse spleen tissue labeling CD4 with ab183685 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    The result showed membrane staining on mouse spleen.

    The nuclear counterstain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183685).

  • CD4 was immunoprecipitated from 1mg of Mouse thymus whole cell lysate with ab183685 at 1/40 dilution.

    Western blot was performed from the immunoprecipitate using ab183685 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

    Lane 1: Mouse thymus whole cell lysate, 10µg (Input).

    Lane 2: ab183685 IP in Mouse thymus whole cell lysate.

    Lane 3: Rabbit IgG,monoclonal [EPR25A] - Isotype Control (ab172730)  instead of ab183685 in Mouse thymus whole cell lysate.

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

    Exposure time: 5 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab183685).

  • This IHC data was generated using the same anti-CD4 antibody clone, EPR19514, in a different buffer formulation (cat# ab183685).

    Immunohistochemical analysis of paraffin-embedded Mouse spleen tissue labeling CD4 with ab183685 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
    Membrane staining on T cells is observed.
    Counter stained with Hematoxylin.
     
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

References

This product has been referenced in:

  • Chute C  et al. Syndecan-1 induction in lung microenvironment supports the establishment of breast tumor metastases. Breast Cancer Res 20:66 (2018). Read more (PubMed: 29976229) »
  • Zhang C  et al. The mTOR signal regulates myeloid-derived suppressor cells differentiation and immunosuppressive function in acute kidney injury. Cell Death Dis 8:e2695 (2017). IHC-P ; Mouse . Read more (PubMed: 28333137) »
See all 5 Publications for this product

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