Recombinant
RabMAb

Recombinant Anti-CD4 antibody [EPR6855] - BSA and Azide free (ab181724)

Overview

  • Product name

    Anti-CD4 antibody [EPR6855] - BSA and Azide free
    See all CD4 primary antibodies
  • Description

    Rabbit monoclonal [EPR6855] to CD4 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IF, Flow Cyt, IHC-Pmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Recombinant fragment within Human CD4 aa 200-400. The exact sequence is proprietary.

  • Positive control

    • FC: Human whole blood IHC-P: Human tonsil, spleen, thymoma, colon, liver WB: THP-1 Negative control: no staining on human cerebrum and pancreas.
  • General notes

    Ab181724 is the carrier-free version of ab133616. 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 <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab181724 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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 ab181724 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
WB Use at an assay dependent concentration. Predicted molecular weight: 51 kDa.

Please check the parent abID, ab133616, for a recommended dilution.

ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval 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

  • Imaging Mass Cytometry™ (IMC™) image of human tonsil tissue stained with Anti-CD4 antibody [EPR6855]. ab181724 (carrier-free antibody, purified) was metal-conjugated using a Maxpar® Antibody Labeling Kit from Fluidigm. Immunostaining was performed according to Fluidigm’s protocols. Briefly, slides were subject to deparaffinization and heat-induced epitope retrieval, followed by overnight incubation at 4°C with an antibody cocktail containing metal-tagged antibodies in blocking buffer. Slides were subsequently washed with 0.2% Triton-X and 1x PBS, counterstained with Cell-ID™ Intercalator-Ir diluted at 1/400 in 1x PBS for 30 min at room temperature, rinsed for 5 min with distilled H2O, and air-dried prior to IMC™ acquisition. IMC™ acquisition was performed using the Fluidigm Hyperion™ Imaging System.

    Imaging Mass Cytometry™, IMC™, Cell-ID™, Hyperion™ and Maxpar® are trademarks of Fluidigm Canada Inc.​​

  • Human peripheral blood lymphocytes stained with unpurified ab133616 (red line). Human whole blood was processed using a modified protocol based on Chow et al, 2005 (PMID: 16080188). In brief, human whole blood was fixed in 4% formaldehyde (methanol-free) for 10 min at 22°C. Red blood cells were then lyzed by the addition of Triton X-100 (final concentration - 0.1%) for 15 min at 37°C. For experimentation, cells were treated with 50% methanol (-20°C) for 15 min at 4°C. Cells were then incubated with the antibody (unpurified ab133616, 1/100 dilution) for 30 min at 4°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 4°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >30,000 total events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. Gating strategy - peripheral blood lymphocytes.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133616).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD4 with purified ab133616 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133616).

  • Anti-CD4 antibody [EPR6855] - BSA and Azide free (ab181724) + THP-1 (human acute monocytic leukemia) whole cell lysate at 10 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051)

    Predicted band size: 51 kDa


    Exposure time: 3 minutes


    This WB data was generated using the same anti-CD4 antibody clone [EPR6855] in a different buffer formulation (cat# ab133616).

    Blocking buffer and concentration: 5% NFDM/TBST

    Diluting buffer and concetration: 5% NFDM/TBST

  • Anti-CD4 antibody [EPR6855] (ab133616)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD4 with ab133616 at a dilution of 1:500. Heat mediated antigen retrieval was performed using AR9 antigen retrieval solution, and microwave treatment for 15 min at 20% power. Anti-Rabbit/Mouse HRP polymer (PerkinElmer Opal Polymer HRP Ms Plus Rb) was used as secondary antibody. Opal tyramide amplification was performed using Opal 520 fluorophore. Counterstained with DAPI stain. Image scanned with Vectra 3.0 and analyzed via Phenochart software.
    This image was courteously provided by Dr. Houssein Abdul Sater, Georgia Cancer Center.

  • Negative control: no staining on human cerebrum.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human cerebrum showing no staining CD4 with purified ab133616 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9 (ab93684). Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133616).

  • Negative control: no staining on human pancreas.

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human pancreas showing no staining CD4 with purified ab133616 at 1/100. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9 (ab93684). Goat Anti-Rabbit IgG H&L (HRP) was used as the secondary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133616).

  • Paraffin-embedded human spleen tissue stained for CD4 using ab133616 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 (ab133616).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thymoma tissue labelling CD4 with purified ab133616 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133616).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling CD4 with ab133616 at a dilution of 1/100.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133616).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling CD4 with unpurified ab133616.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133616).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling CD4 with unpurified ab133616.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133616).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thymoma tissue labelling CD4 with unpurified ab133616.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab133616).

  • This IHC data was generated using the same anti-CD4 antibody clone [EPR6855] in a different buffer formulation (cat# ab133616).

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human tonsil tissue labelling CD4 with unpurified ab133616 at a dilution of 1/100.

References

ab181724 has not yet been referenced specifically in any publications.

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