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    products/primary-antibodies/cd4-antibody-sp35-ab213215.pdf

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RecombinantRabMAb

Recombinant Anti-CD4 antibody [SP35] (ab213215)

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  • SDS
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Multiplex immunohistochemistry - Anti-CD4 antibody [SP35] (ab213215)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
  • Flow Cytometry - Anti-CD4 antibody [SP35] (ab213215)
  • Multiplex immunohistochemistry - Anti-CD4 antibody [SP35] (ab213215)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
  • Flow Cytometry - Anti-CD4 antibody [SP35] (ab213215)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
  • Anti-CD4 antibody [SP35] (ab213215)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [SP35] to CD4
  • Suitable for: IHC-P, Flow Cyt, mIHC
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alkaline Phosphatase APC Carrier Free HRP PE

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Overview

  • Product name

    Anti-CD4 antibody [SP35]
    See all CD4 primary antibodies
  • Description

    Rabbit monoclonal [SP35] to CD4
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, Flow Cyt, mIHCmore details
  • Species reactivity

    Reacts with: Human
    Predicted to work with: Chimpanzee
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • Flow Cyt: THP-1 and Jurkat cells. IHC-P: Human Hodgkin's lymphoma and tonsil tissue.
  • General notes

    This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.

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: 7.60
    Preservative: 0.1% Sodium azide
    Constituents: PBS, 1% BSA
  • Concentration information loading...
  • Purity

    Protein A/G purified
  • Purification notes

    Purified from TCS by protein A/G.
  • Clonality

    Monoclonal
  • Clone number

    SP35
  • 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 [SP35], prediluted (ab101530)
    • Anti-CD4 antibody [SP35] - BSA and Azide free (ab238798)
    • Alexa Fluor® 568 Anti-CD4 antibody [SP35] (ab312782)
  • 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 ab213215 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
IHC-P (3)
1/50.

Deparaffinize slides using xylene or xylene alternative and graded alcohols. Boil tissue section in EDTA buffer for 10 min followed by cooling at room temperature for 20 min.  Incubate with primary antibody for 30 minutes at room temperature. Slides must be washed in between steps.  Rinse slides with PBS/0.05% Tween. 

Flow Cyt
1/20 - 1/100.

Incubate with primary antibody for 30 minutes at 4°C.

mIHC
Use at an assay dependent concentration.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2)

Notes
IHC-P
1/50.

Deparaffinize slides using xylene or xylene alternative and graded alcohols. Boil tissue section in EDTA buffer for 10 min followed by cooling at room temperature for 20 min.  Incubate with primary antibody for 30 minutes at room temperature. Slides must be washed in between steps.  Rinse slides with PBS/0.05% Tween. 

Flow Cyt
1/20 - 1/100.

Incubate with primary antibody for 30 minutes at 4°C.

mIHC
Use at an assay dependent concentration.

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2)

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: 450124 Chimpanzee
    • Entrez Gene: 920 Human
    • Omim: 186940 Human
    • SwissProt: P16004 Chimpanzee
    • SwissProt: P01730 Human
    • Unigene: 631659 Human
    • 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

    • Multiplex immunohistochemistry - Anti-CD4 antibody [SP35] (ab213215)
      Multiplex immunohistochemistry - Anti-CD4 antibody [SP35] (ab213215)

      This data was developed using ab238798 , the same antibody clone in a different buffer formulation.

       

      Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human spleen tissue labeling CD8 alpha with ab245118 at 1/500 dilution, CD4 with ab238798 at 1/500, and CD19 with ab237772 at 1/5000 dilution.

      Panel A: merged staining of anti-CD8 alpha (magenta; Opal™690), anti-CD4 (green; Opal™520) and anti-CD19 (red; Opal™570) on human spleen.
      Panel B: anti-CD8 alpha stained on cytotoxic T cells.
      Panel C: anti-CD4 stained on T helper cells.
      Panel D: anti-CD19 stained on B cells.

      Sections  were treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins before antibody incubation. The section was incubated in three rounds of staining: in the order of abab245118 for 30 mins, then ab238798 and ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

      DAPI was used as a nuclear counterstain.

      The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human Hodgkin lymphoma tissue sections labeling CD4 with ab213215 at 1/50 dilution (3.1 µg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on the human Hodgkin's lymphoma, performed on a Leica Biosystems BOND™ RX instrument.
      The section was incubated with ab213215 for 30 mins at room temperature.

    • Flow Cytometry - Anti-CD4 antibody [SP35] (ab213215)
      Flow Cytometry - Anti-CD4 antibody [SP35] (ab213215)

      Flow cytometry analysis of THP-1 (human acute monocytic leukemia) labeling CD4 with purified ab213215 at 1/20 dilution (7.75 µg/ml) (red).

      Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as a secondary antibody. Isotype control - Rabbit monoclonal IgG (ab172730) (Black). Unlableled control - Unlabeled cells (blue).

    • Multiplex immunohistochemistry - Anti-CD4 antibody [SP35] (ab213215)
      Multiplex immunohistochemistry - Anti-CD4 antibody [SP35] (ab213215)

      This data was developed using ab238798 , the same antibody clone in a different buffer formulation.

       

      Multiplex immunohistochemistry analysis of formalin/PFA-fixed paraffin-embedded Human tonsil tissue labeling CD8 alpha with ab245118 at 1/500 dilution, CD4 with ab238798 at 1/500, and CD19 with ab237772 at 1/5000 dilution.

      Panel A: merged staining of anti-CD8 alpha (magenta; Opal™690), anti-CD4 (green; Opal™520) and anti-CD19 (red; Opal™570) on human tonsil.
      Panel B: anti-CD8 alpha stained on cytotoxic T cells.
      Panel C: anti-CD4 stained on T helper cells.
      Panel D: anti-CD19 stained on B cells.

      Sections  were treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins before antibody incubation. The section was incubated in three rounds of staining: in the order of abab245118 for 30 mins, then ab238798 and ab237772 for 10 mins at room temperature. Each round was followed by a separate fluorescent tyramide signal amplification system.

      DAPI was used as a nuclear counterstain.

      The immunostaining was performed on a Leica Biosystems BOND® RX instrument with an Opal™ 4-color kit. Image acquisition was performed with Leica SP8 confocal microscope.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)Image is from Lemos MP et al., PLoS One 9(9). Fig 5A,; doi: 10.1371/journal.pone.0108954 Reproduced under the Creative Commons license https://creativecommons.org/publicdomain/zero/1.0/

      Foreskin epidermis at 10× magnification stained with CCR5 (pseudocolored green), CD4 (pseudocolored red; ab213215) and SYTOX Orange (pseudocolored blue) for nuclear identification. All fields from ∼26.5 mm2 sections were used for analysis.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human tonsil tissue sections labeling CD4 with ab213215 at 1/50 dilution (3.1 µg/ml). Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 10mins. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used as the secondary antibody. Hematoxylin was used as a counterstain. Positive staining on the human tonsil, performed on a Leica Biosystems BOND™ RX instrument.
      The section was incubated with ab213215 for 30 mins at room temperature.

    • Flow Cytometry - Anti-CD4 antibody [SP35] (ab213215)
      Flow Cytometry - Anti-CD4 antibody [SP35] (ab213215)

      Flow cytometric analysis of Jurkat (Human T cell leukemia cell line from peripheral blood) cells labeling CD4 with ab213215 at 1/100 dilution (green), compared with a rabbit IgG negative control (blue).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)

      Immunohistochemical analysis of formalin-fixed, paraffin-embedded human tonsil tissue labeling CD4 with ab213215 at 1/50 dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)

      Immunohistochemical analysis of formalin-fixed, paraffin-embedded human tonsil tissue labeling CD4 with ab213215 at 1/50 dilution.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [SP35] (ab213215)

      Tissue Microarrays stained for "Anti-CD4 antibody [SP35]” using "ab213215" in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 10 minutes. The sections were incubated with ab213215 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

    • Anti-CD4 antibody [SP35] (ab213215)
      Anti-CD4 antibody [SP35] (ab213215)

    Protocols

    • Flow cytometry protocols
    • Immunohistochemistry protocols

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (17)

    Publishing research using ab213215? Please let us know so that we can cite the reference in this datasheet.

    ab213215 has been referenced in 17 publications.

    • Masuda K  et al. Multiplexed single-cell analysis reveals prognostic and nonprognostic T cell types in human colorectal cancer. JCI Insight 7:N/A (2022). PubMed: 35192548
    • Johnson BE  et al. An omic and multidimensional spatial atlas from serial biopsies of an evolving metastatic breast cancer. Cell Rep Med 3:100525 (2022). PubMed: 35243422
    • Ono Y  et al. Relationship between consolidation tumor ratio and tumor-infiltrating lymphocytes in small-sized lung adenocarcinoma. Thorac Cancer 13:2134-2141 (2022). PubMed: 35791738
    • Lai Y  et al. Iron controls T helper cell pathogenicity by promoting glucose metabolism in autoimmune myopathy. Clin Transl Med 12:e999 (2022). PubMed: 35917405
    • Nguyen T  et al. Multiplex Immunohistochemistry Analysis of Melanoma Tumor-Infiltrating Lymphocytes. Methods Mol Biol 2265:557-572 (2021). PubMed: 33704740
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-3 of 3 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD4 antibody [SP35]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (Human Tissue sections (Human Xenograft))
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 1 mM EDTA Buffer + 0.05% Tween20 , pH 8.0
    Permeabilization
    No
    Specification
    Human Tissue sections (Human Xenograft)
    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.

    MS. Loreen Weichert

    Verified customer

    Submitted Mar 11 2022

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD4 antibody [SP35]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Monkey Tissue sections (Skin)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: buffer pH9
    Permeabilization
    No
    Specification
    Skin
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · 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.

    Abcam user community

    Verified customer

    Submitted Nov 10 2021

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-CD4 antibody [SP35]

    Inconclusive
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Dog Tissue sections (lymph node)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: buffer pH9
    Permeabilization
    No
    Specification
    lymph node
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 10% · 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.

    Abcam user community

    Verified customer

    Submitted Nov 10 2021

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com

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