Overview

  • Product name
    Anti-CD4 antibody [EPR19514]
    See all CD4 primary antibodies
  • Description
    Rabbit monoclonal [EPR19514] to CD4
  • Host species
    Rabbit
  • Tested applications
    Suitable for: IP, IHC-P, IHC-Fr, WBmore details
  • Species reactivity
    Reacts with: Mouse
  • Immunogen

    Recombinant fragment within Mouse CD4 aa 1-400. The exact sequence is proprietary.
    Database link: P06332

  • Positive control
    • WB: Mouse thymus lysate. IHC-P: Mouse spleen, colon and thymus tissues. IHC-Fr: Mouse spleen tissue. IP: Mouse thymus whole cell lysate.
  • 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 ab183685 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 1/40.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IHC-Fr 1/200.
WB 1/1000. Detects a band of approximately 51 kDa (predicted molecular weight: 51 kDa).

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

  • 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.
  • 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.

  • All lanes : Anti-CD4 antibody [EPR19514] (ab183685) at 1/2000 dilution

    Lane 1 : Mouse thymus lysate
    Lane 2 : Mouse liver tissue lysate
    Lane 3 : Mouse brain tissue lysate
    Lane 4 : Mouse kidney tissue lysate
    Lane 5 : Raw264.7 (mouse abelson murine leukemia virus-induced tumor) whole cell lysate
    Lane 6 : NIH/3T3 (mouse embryo) 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: 51 kDa
    Observed band size: 51 kDa



    The expression profile observed is consistent with what has been described in the literature (PMID: 2155425).

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1:3 seconds; Lanes 2,3,4,5 and 6:30 seconds

     

  • 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.

  • 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.
  • 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.

    See Abreview

  • 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.

  • Formaldehyde-fixed, paraffin-embedded mouse spleen tissue stained for CD4 using ab183685 at 1/2000 dilution in immunohistochemical analysis, followed by Goat anti Rabbit IgG Biotin.

    See Abreview

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

    See Abreview

References

This product has been referenced in:
  • Camilio KA  et al. Combining the oncolytic peptide LTX-315 with doxorubicin demonstrates therapeutic potential in a triple-negative breast cancer model. Breast Cancer Res 21:9 (2019). Read more (PubMed: 30670061) »
  • Hosomi S  et al. Myeloid-Derived Suppressor Cells Infiltrate the Brain and Suppress Neuroinflammation in a Mouse Model of Focal Traumatic Brain Injury. Neuroscience 406:457-466 (2019). Read more (PubMed: 30880103) »
See all 19 Publications for this product

Customer reviews and Q&As

1-10 of 12 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Spleen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Leica ER2 solution
Permeabilization
No
Specification
Spleen
Fixative
Formaldehyde

Zohar Gavish

Verified customer

Submitted May 13 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (B16F10 (melanoma) syngeneic mouse tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA
Permeabilization
Yes - 3% H2O2
Specification
B16F10 (melanoma) syngeneic mouse tumor
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Apr 30 2019

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (lungs)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer pH 6
Permeabilization
No
Specification
lungs
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Nov 28 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (brain tissue)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tris/EDTA ph 9
Permeabilization
No
Specification
brain tissue
Blocking step
Normal Goat Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 26°C
Fixative
Formaldehyde

Herr Dr. Markus Kipp

Verified customer

Submitted Aug 21 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Eo771 mammary tumor)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate Buffer, Vector laboratories(H-3300)
Permeabilization
Yes - 0.2% Triton-X
Specification
Eo771 mammary tumor
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
Fixative
Formalin

Anugraha Rajagopalan

Verified customer

Submitted Apr 11 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Thymus)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Bond TM Epitope Retrieval 2-1L (Leika Biosystems)
Permeabilization
No
Specification
Thymus
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Mar 27 2018

Application
Western blot
Sample
Mouse Cell lysate - whole cell (mouse hepatocytes)
Gel Running Conditions
Reduced Denaturing
Loading amount
25 µg
Specification
mouse hepatocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jan 10 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Colon)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: pH 9.0 EDTA
Permeabilization
No
Specification
Colon
Blocking step
Protein Block ab64226 as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: RT°C
Fixative
Formaldehyde

Abcam user community

Verified customer

Submitted Dec 07 2017

Application
Immunohistochemistry (Frozen sections)
Sample
Mouse Tissue sections (Liver)
Permeabilization
Yes - 0.1% tween, trition 0.3% in PBS
Specification
Liver
Blocking step
BSA as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 4°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 30 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Spleen)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citric acid
Permeabilization
No
Specification
Spleen
Blocking step
BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
Fixative
Formaldehyde

Mr. Carl Hobbs

Verified customer

Submitted Mar 27 2017

1-10 of 12 Abreviews or Q&A

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