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    cd4-antibody-cal4-ab237722.pdf

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Immunology Adaptive Immunity T Cells CD
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RecombinantRabMAb

Recombinant Anti-CD4 antibody [CAL4] (ab237722)

  • Datasheet
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [CAL4] (ab237722)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [CAL4] (ab237722)
  • Western blot - Anti-CD4 antibody [CAL4] (ab237722)
  • Immunohistochemistry (Frozen sections) - Anti-CD4 antibody [CAL4] (ab237722)
  • Flow Cytometry (Intracellular) - Anti-CD4 antibody [CAL4] (ab237722)
  • Immunoprecipitation - Anti-CD4 antibody [CAL4] (ab237722)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [CAL4] (ab237722)
  • Anti-CD4 antibody [CAL4] (ab237722)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [CAL4] to CD4
  • Suitable for: WB, IHC-Fr, IP, Flow Cyt (Intra), IHC-P
  • Reacts with: Mouse, Rat

Conjugates logo Related conjugates and formulations

Carrier Free

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Overview

  • Product name

    Anti-CD4 antibody [CAL4]
    See all CD4 primary antibodies
  • Description

    Rabbit monoclonal [CAL4] to CD4
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-Fr, IP, Flow Cyt (Intra), IHC-Pmore details
    Unsuitable for: ICC/IF
  • Species reactivity

    Reacts with: Mouse, Rat
  • Immunogen

    Synthetic peptide within Mouse CD4 aa 417-457. The exact sequence is proprietary.
    Database link: P06332

  • Positive control

    • IHC-P: Mouse and rat spleen tissue. WB: Mouse and rat thymus, spleen and lymph node lysate. IHC-Fr: Mouse spleen tissue. Flow Cyt (intra): Mouse splenocytes. IP: Mouse thymus lysate.

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

    pH: 7.2
    Preservative: 0.05% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Purification notes

    Purity >99%.
  • Clonality

    Monoclonal
  • Clone number

    CAL4
  • Isotype

    IgG
  • Research areas

    • Immunology
    • Adaptive Immunity
    • T Cells
    • CD
    • Stem Cells
    • Hematopoietic Progenitors
    • Lymphoid
    • T Lymphocytic Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Dendritic Cell Lineage
    • Stem Cells
    • Hematopoietic Progenitors
    • Myeloid
    • Monocytic Lineage
    • Immunology
    • Adaptive Immunity
    • Regulatory T Cells
    • Immunology
    • Immune System Diseases
    • Antiviral Signaling
    • HIV-related

Associated products

  • Alternative Versions

    • Anti-CD4 antibody [CAL4] - BSA and Azide free (ab251608)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab237722 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
1/1000. Predicted molecular weight: 51 kDa.
IHC-Fr
1/100.

Perform heat mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 and 0.05% Tween-20).

IP
1/30.
Flow Cyt (Intra)
1/40.
IHC-P (1)
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Notes
WB
1/1000. Predicted molecular weight: 51 kDa.
IHC-Fr
1/100.

Perform heat mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 and 0.05% Tween-20).

IP
1/30.
Flow Cyt (Intra)
1/40.
IHC-P
1/4000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Application notes
Is unsuitable for ICC/IF.

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.
  • Target information above from: UniProt accession P01730 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 12504 Mouse
    • Entrez Gene: 24932 Rat
    • SwissProt: P06332 Mouse
    • SwissProt: P05540 Rat
    • Unigene: 2209 Mouse
    • Unigene: 10748 Rat
    • 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

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [CAL4] (ab237722)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [CAL4] (ab237722)

      Immunohistochemical analysis of paraffin-embedded mouse spleen tissue labeling CD4 with ab237722 at 1/4000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on mouse spleen. The section was incubated with ab237722 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use 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 mins.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [CAL4] (ab237722)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [CAL4] (ab237722)

      Immunohistochemical analysis of paraffin-embedded rat spleen tissue labeling CD4 with ab237722 at 1/4000 dilution, followed by ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on rat spleen. The section was incubated with ab237722 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ready to use 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 mins.

    • Western blot - Anti-CD4 antibody [CAL4] (ab237722)
      Western blot - Anti-CD4 antibody [CAL4] (ab237722)
      All lanes : Anti-CD4 antibody [CAL4] (ab237722) at 1/1000 dilution

      Lane 1 : Mouse thymus lysate
      Lane 2 : Mouse spleen lysate
      Lane 3 : Mouse lymph node lysate
      Lane 4 : Rat thymus lysate
      Lane 5 : Rat spleen lysate
      Lane 6 : Rat lymph node 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



      Blocking/Dilution buffer: 5% NFDM/TBST.

    • Immunohistochemistry (Frozen sections) - Anti-CD4 antibody [CAL4] (ab237722)
      Immunohistochemistry (Frozen sections) - Anti-CD4 antibody [CAL4] (ab237722)

      Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen mouse spleen tissue labeling CD4 with ab237722 at 1/100 dilution (green), followed by ab150077 AlexaFluor®488 Goat anti-Rabbit secondary at a 1/1,000 dilution. Positive staining on T cell surface (PMID: 26994219) is observed. Counterstained with DAPI (blue). Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab150077 AlexaFluor®488 Goat anti-Rabbit used at a 1/1,000 dilution. Heat mediated antigen retrieval using sodium citrate buffer (10 mM citrate pH 6.0 and 0.05% Tween-20).

    • Flow Cytometry (Intracellular) - Anti-CD4 antibody [CAL4] (ab237722)
      Flow Cytometry (Intracellular) - Anti-CD4 antibody [CAL4] (ab237722)

      Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed, 0.1% Tween-20 permeabilized mouse splenocytes labeling CD4 with ab237722 at 1/40 (right panel) compared with a Rabbit monoclonal IgG (ab172730) (left panel). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody. Cells were surface stained with anti-CD4 conjugated to Alexa Fluor® 647. Then fixed with 2% PFA for 10min and intracellularly stained with rabbit IgG (Left) or ab237722 (Right). They recognize the same cell population. 

       

       

    • Immunoprecipitation - Anti-CD4 antibody [CAL4] (ab237722)
      Immunoprecipitation - Anti-CD4 antibody [CAL4] (ab237722)

      CD4 was immunoprecipitated from 0.35 mg mouse thymus lysate with ab237722 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab237722 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used at 1/5000 dilution.
      Lane 1: Mouse thymus lysate 10 µg (Input).
      Lane 2: ab237722 IP in mouse thymus lysate .
      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab237722 in mouse thymus lysate .
      Blocking/Dilution buffer: 5% NFDM/TBST.
      Exposure time: 30 seconds.

      102kDa band represents CD4 dimer based on literature. (PMID: 20739067).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [CAL4] (ab237722)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [CAL4] (ab237722)

      Formalin-fixed, paraffin-embedded mouse spleen tissue stained for CD4 using ab237722 at 0.5 µg/ml in immunohistochemical analysis.

    • Anti-CD4 antibody [CAL4] (ab237722)
      Anti-CD4 antibody [CAL4] (ab237722)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (4)

    Publishing research using ab237722? Please let us know so that we can cite the reference in this datasheet.

    ab237722 has been referenced in 4 publications.

    • Irastorza-Lorenzo A  et al. Evaluation of Marine Agarose Biomaterials for Tissue Engineering Applications. Int J Mol Sci 22:N/A (2021). PubMed: 33672027
    • Chen KT  et al. Neuronavigation-guided focused ultrasound for transcranial blood-brain barrier opening and immunostimulation in brain tumors. Sci Adv 7:N/A (2021). PubMed: 33547073
    • Zhu D  et al. Minimally invasive delivery of therapeutic agents by hydrogel injection into the pericardial cavity for cardiac repair. Nat Commun 12:1412 (2021). PubMed: 33658506
    • Ortiz-Arrabal O  et al. Generation and Evaluation of Novel Biomaterials Based on Decellularized Sturgeon Cartilage for Use in Tissue Engineering. Biomedicines 9:N/A (2021). PubMed: 34356839

    Customer reviews and Q&As

    Show All Reviews Q&A
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    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD4 antibody [CAL4]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Tonsille)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: high Target retrival solution
    Permeabilization
    No
    Specification
    Tonsille
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 5µg/mL · Temperature: 25°C
    Fixative
    Formaldehyde
    Read More
    The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

    FRAU. Ines Kleiber-Schaaf

    Verified customer

    Submitted Aug 07 2020

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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