PE Anti-CD4 antibody [MEM-241] (ab18282)

Reviews (6)Q&A (6)References (4)
Flow Cytometry - PE Anti-CD4 antibody [MEM-241] (ab18282)

    Key features and details

    • PE Mouse monoclonal [MEM-241] to CD4
    • Suitable for: Flow Cyt
    • Reacts with: Human
    • Conjugation: PE. Ex: 488nm, Em: 575nm
    • Isotype: IgG1

    Overview

    • Product name

      PE Anti-CD4 antibody [MEM-241]
      See all CD4 primary antibodies

    • Description

      PE Mouse monoclonal [MEM-241] to CD4

    • Host species

      Mouse

    • Conjugation

      PE. Ex: 488nm, Em: 575nm

    • Tested applications

      Suitable for: Flow Cytmore details

    • Species reactivity

      Reacts with: Human

    • Immunogen

      CD4 Fusion protein (Human).

    • General notes

      The purified antibody is conjugated with R-Phycoerythrin (R-PE) under optimum conditions. The conjugate is purified by size-exclusion chromatography and adjusted for direct use.

      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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

      If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

    Properties

    Applications

    The Abpromise guarantee

    Our Abpromise guarantee covers the use of ab18282 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
    Flow Cyt (5)
    Use at an assay dependent concentration.

    Use 5-20µl for 100µl whole human blood cells.

     

     

     

    ab91357 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

     
    Notes
    Flow Cyt
    Use at an assay dependent concentration.

    Use 5-20µl for 100µl whole human blood cells.

     

     

     

    ab91357 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

     

    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

      • Flow Cytometry - PE Anti-CD4 antibody [MEM-241] (ab18282)
        Flow Cytometry - PE Anti-CD4 antibody [MEM-241] (ab18282)

        ab18282 staining CD4 in human T cells by Flow Cytometry . 

      References (4)

      Publishing research using ab18282? Please let us know so that we can cite the reference in this datasheet.

      ab18282 has been referenced in 4 publications.

      • Kim S  et al. Improved Enumeration of Weakly Fluorescent CD4+ T-lymphocytes by Confining Cells in a Spinning Sample Cartridge with a Helical Minichannel. Micromachines (Basel) 11:N/A (2020). PubMed: 32630535
      • Hernández-Castro JA  et al. Two-level submicron high porosity membranes (2LHPM) for the capture and release of white blood cells (WBCs). Lab Chip 19:589-597 (2019). PubMed: 30648711
      • Dieckmann NM  et al. Munc18-2 is required for Syntaxin 11 Localization on the Plasma Membrane in Cytotoxic T-Lymphocytes. Traffic 16:1330-41 (2015). PubMed: 26771955
      • Seyedi S  et al. The Effects of Isoproterenol and Propranolol on Cytokine Profile Secretion by Cultured Tumor-infiltrating Lymphocytes Derived from Colorectal Cancer Patients. Cell J 13:281-9 (2012). PubMed: 23507624

      Customer reviews and Q&As

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      1-10 of 12 Abreviews or Q&A

      Flow Cytometry abreview for Anti-CD4 antibody [MEM-241] (Phycoerythrin)

       Poor
      Abreviews
      abreview image
      Application
      Flow Cytometry
      Sample
      Rat Cell (Splenocytes)
      Specification
      Splenocytes
      Preparation
      Cell harvesting/tissue preparation method: harvesting of spleen
      Sample buffer: PBS/BSA 1%
      Permeabilization
      No
      Gating Strategy
      alive lymphocytes
      Read More

      Abcam user community

      Verified customer

      Submitted Nov 13 2012

      Flow Cytometry abreview for Anti-CD4 antibody [MEM-241] (Phycoerythrin)

       Poor
      Abreviews
      abreview image
      Application
      Flow Cytometry
      Sample
      Mouse Cell (Splenocytes)
      Specification
      Splenocytes
      Preparation
      Cell harvesting/tissue preparation method: harvesting of spleen
      Sample buffer: PBS/BSA 1%
      Permeabilization
      No
      Gating Strategy
      alive lymphocytes
      Read More

      Abcam user community

      Verified customer

      Submitted Nov 13 2012

      Flow Cytometry abreview for Anti-CD4 antibody [MEM-241] (Phycoerythrin)

       Good
      Abreviews
      abreview image
      Application
      Flow Cytometry
      Sample
      Monkey Cell (leucocytes from whole blood)
      Specification
      leucocytes from whole blood
      Preparation
      Cell harvesting/tissue preparation method: after red blood cell lysis
      Sample buffer: PBS/BSA 1%
      Fixation
      Paraformaldehyde
      Permeabilization
      No
      Gating Strategy
      alive lymphocytes
      Read More

      Abcam user community

      Verified customer

      Submitted Nov 13 2012

      Flow Cytometry abreview for Anti-CD4 antibody [MEM-241] (Phycoerythrin)

       Good
      Abreviews
      abreview image
      Application
      Flow Cytometry
      Sample
      Human Cell (leucocytes from whole blood)
      Specification
      leucocytes from whole blood
      Preparation
      Cell harvesting/tissue preparation method: after red blood cell lysis
      Sample buffer: PBS/BSA 1%
      Fixation
      Paraformaldehyde
      Permeabilization
      No
      Gating Strategy
      alive cells
      Read More

      Abcam user community

      Verified customer

      Submitted Nov 12 2012

      Flow Cytometry abreview for Anti-CD4 antibody [MEM-241] (Phycoerythrin)

       Poor
      Abreviews
      abreview image
      Application
      Flow Cytometry
      Sample
      Dog Cell (from whole blood)
      Specification
      from whole blood
      Preparation
      Cell harvesting/tissue preparation method: after red blood cell lysis
      Sample buffer: PBS/BSA 1%
      Fixation
      Paraformaldehyde
      Permeabilization
      No
      Gating Strategy
      alive lymphocytes
      Read More

      Abcam user community

      Verified customer

      Submitted Nov 07 2012

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD4 antibody [MEM-241] (Phycoerythrin)

       Inconclusive
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
      Sample
      Mouse Tissue sections (Thymus)
      Specification
      Thymus
      Fixative
      Paraformaldehyde
      Antigen retrieval step
      Heat mediated - Buffer/Enzyme Used: Citrate buffer PH6.0
      Permeabilization
      No
      Blocking step
      Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
      Read More

      Dr. Ruifeng Teng

      Verified customer

      Submitted Apr 30 2012

      Question

      Thank you for your advices. Actually, as the whole blood staining could induce some artefacts, we have performed some additional controls in those experiments :

      We have used the antibody directly in the blood (before fixation and lysis) at a 1/10 dilution with a 2h RT incubation. This was performed to be sure that the PFA does not alter some important epitope. In parallel, we have done experiments were the antibody incubation was performed after fixation and red blood cell lysis.

      No staining was observed in all situations.

      Also, please note that we have used the same protocol for experiment with cynomologus monkeys in a context of a preclinical study and the antibody worked perfectly well in the very same conditions.

      For all those reasons, we believed that the antibody did not work probably because of lack of cross-reactivity to the mouse/rat orthologue.

      Read More
      Answer

      Thank you so much for the additional information.
      Yes, this probably explains why you did not get any signal with mouse and rat samples, but got signal with monkey.
      Thank you so much for submitting your reviews!

      Read More

      Question

      ReviewID: xxx AbID: 18282, Anti-CD4 antibody [MEM-241] (Phycoerythrin) Rating: Poor Image: Anti-CD4 antibody [MEM-241] (Phycoerythrin) for Flow Cytometry (Mouse) Sample: Species: Mouse Type: Cell Specification: Splenocytes Preparation: Cell harvesting/tissue preparation method: harvesting of spleen
      Sample buffer: PBS/BSA 1% Application: Fixation: Permeabilization: No Gating Strategy: alive lymphocytes Other detail: Dilution: 1/100 Incubation time: 1 hour at 4°C Diluent: PBS/BSA 1% Secondary Antibody: Name: None used Conjugation: Additional Data:

      Read More
      Answer

      Thank you for submitting your Abreviews for ab18282. As you may have noticed, your reviews has now been published on our website.
      Since you obtained poor results using the antibody in the untested species (mouse and rat), we would like to follow up on this to see if we can possibly improve the results you are seeing with this antibody.
      Based on the information in your review, I feel that altering the following step in your protocol may help improve your results:
      - longer incubation time with the primary antibody
      - using more antibody
      However, alternatively, this could also mean that the crossreactivity with mouse and rat is poor due to low homology.
      If you decide to follow these suggestions, please let me know if they are helpful.
      I wish you good luck with your research.

      Read More

      Question

      Thank you for your advices. Actually, as the whole blood staining could induce some artefacts, we have performed some additional controls in those experiments :

      We have used the antibody directly in the blood (before fixation and lysis) at a 1/10 dilution with a 2h RT incubation. This was performed to be sure that the PFA does not alter some important epitope. In parallel, we have done experiments were the antibody incubation was performed after fixation and red blood cell lysis.

      No staining was observed in all situations.

      Also, please note that we have used the same protocol for experiment with cynomologus monkeys in a context of a preclinical study and the antibody worked perfectly well in the very same conditions.

      For all those reasons, we believed that the antibody did not work probably because of lack of cross-reactivity to the dog orthologue.

      Read More
      Answer

      Thank you for the additional information and thank you for submitting your Abreviews.

      Read More

      Question

      ReviewID: xxx AbID: 18282, Anti-CD4 antibody [MEM-241] (Phycoerythrin) Rating: Poor Image: Anti-CD4 antibody [MEM-241] (Phycoerythrin) for Flow Cytometry (Dog) Sample: Species: Dog Type: Cell Specification: from whole blood Preparation: Cell harvesting/tissue preparation method: after red blood cell lysis
      Sample buffer: PBS/BSA 1% Application: Application: Fixation: Paraformaldehyde Permeabilization: No Gating Strategy: alive lymphocytes Other detail: Dilution: 1/100 Incubation time: 1 hour at 4°C Diluent: PBS/BSA 1% Secondary Antibody: Name: None used Conjugation: Additional Data:

      Read More
      Answer

      Thank you for submitting an Abreview for ab18282. As you may have noticed, your review has now been published on our website.
      Since you obtained poor results using the antibody in an untested species (dog), we would like to follow up on this to see if we can possibly improve the results you are seeing with this antibody.
      Based on the information in your review, I feel that altering the following step in your protocol may help improve your results:
      - fix with PFA after incubation with the primary antibody - if you haven't done that already.
      - incubate longer with the primary antibody (e.g. 2-3 h at RT or overnight at 4 degree)
      It could also just be the case that the antibody does not work for dog samples due to low homology.
      If you decide to follow these suggestions, please let me know if they are helpful.
      I wish you good luck with your research.

      Read More

      1-10 of 12 Abreviews or Q&A

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