Recombinant Anti-CD4 antibody [EPR19514] (ab183685)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19514] to CD4
- Suitable for: IP, IHC-P, IHC-Fr, WB
- Reacts with: Mouse
Overview
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Product name
Anti-CD4 antibody [EPR19514]
See all CD4 primary antibodies -
Description
Rabbit monoclonal [EPR19514] to CD4 -
Host species
Rabbit -
Tested Applications & Species
Application Species IHC-Fr MouseIHC-P MouseIP MouseWB Mouse -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: Mouse thymus, liver, brain and kidney tissue lysates; Raw264.7 and NIH/3T3 cell lysates IHC-P: Mouse spleen, colon, brain, cerebrum, lung and thymus tissues; mammary tumor tissue IHC-Fr: Mouse spleen and liver tissue. IP: Mouse thymus whole cell lysate.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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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: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19514 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
Application | Species |
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IHC-Fr |
Mouse
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IHC-P |
Mouse
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IP |
Mouse
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WB |
Mouse
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Application | Abreviews | Notes |
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IP |
1/40.
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IHC-P | (14) |
1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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IHC-Fr | (4) |
1/200.
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WB | (1) |
1/1000. Detects a band of approximately 51 kDa (predicted molecular weight: 51 kDa).
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Notes |
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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
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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
modificationsPalmitoylation 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
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Database links
- Entrez Gene: 12504 Mouse
- SwissProt: P06332 Mouse
- Unigene: 2209 Mouse
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Alternative names
- CD 4 antibody
- CD4 (L3T4) antibody
- CD4 antibody
see all
Images
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (ab183685)Verified customer
Formaldehyde-fixed, non-permeabilized mouse colon tissue stained for CD4 with ab183685 (30 mins at a 1/500 dilution) in immunohistochemical analysis. A Rabbit polyclonal HRP conjugate was used as the secondary.
Heat mediated antigen retrieval buffer/enzyme used: pH 9.0 EDTA.
Blocking step: 1% ab64226 for 10 mins at RT.
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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 kDaThe 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
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IHC image of CD4 staining in a section of frozen normal Mouse Spleen.
The section was fixed using 10% formaldehyde in 1XPBS for 10 minutes. No antigen retrieval step was performed prior to staining. Non-specific protein-protein interactions were then blocked in TBS containing 0.025% (v/v) Triton X-100, 0.3M glycine and 1% (w/v) BSA for 1h at room temperature. The section was then incubated overnight at +4°C in TBS containing 0.025% (v/v) Triton X-100 and 1% (w/v) BSA with ab183685 (Rabbit monoclonal [EPR19514] to CD4) at 1/200 and ab243840 at 1µg/ml, to show the distinct staining of B cells and T cells. The section was then incubated with ab150165 (Goat Anti-Rat IgG H&L (Alexa Fluor® 488) preabsorbed, (Shown in green) 1/1000) and ab150080 (Goat Anti-Rabbit IgG H&L (Alexa Fluor®594) (Shown in red) 1/1000) for 1 hour at room temperature. The secondary-only control insert image is taken from an identical assay without primary antibody. DAPI was used to stain the cell nuclei (blue). The section was then mounted using Fluoromount®.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
For IHC staining systems (automated and non-automated), customers should optimize variable parameters such as antibody concentrations and incubation times.
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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.
Antigen retrieval: Heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20).
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Paraformaldehyde-fixed, 0.1% Tween 0.3% Triton in PBS permeabilized mouse liver tissue stained for CD4 with ab183685 (12 hours, 4°C at a 1/100 dilution) in immunohistochemical analysis. A Donkey anti Rabbit IgG (H+L) AlexaFluor®647 was used as the secondary at a 1/500 dilution (red).
Blocking step: 1% BSA for 12 hours at 4°C.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (ab183685)Anugraha Rajagopalan
Formalin-fixed, 0.2% Triton-X permeabilized mouse Eo771 mammary tumor tissue stained for CD4 with ab183685 (18 hours, 4°C at a 1/1000 dilution) in immunohistochemical analysis. A Goat anti Rabbit IgG (H&L) polyclonal AlexaFluor®488 conjugate was used as the secondary (green).
Heat mediated antigen retrieval buffer/enzyme used: Citrate buffer.
Blocking step: 4% BSA for 1 hour at 25°C.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (ab183685)Herr Dr. Markus Kipp
Formaldehyde-fixed, non-permeabilized mouse brain tissue stained for CD4 with ab183685 (14 hours, 4°C at a 1/1000 dilution) in immunohistochemical analysis. A Goat anti Rabbit IgG monoclonal HRP conjugate was used as the secondary at a 1/200 dilution.
Blocking step: 5% normal goat serum for 1 hour at 26°C.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (ab183685)Verified customer
Formaldehyde-fixed, paraffin-embedded mouse lung tissue stained for CD4 with ab183685 (18 hours, 4°C at a 1/1000 dilution) in immunohistochemical analysis. A Goat anti Rabbit IgG polyclonal HRP conjugate was used as the secondary at a 1/250 dilution (red).
Heat mediated antigen retrieval buffer/enzyme used: Citrate buffer pH 6.0.
Blocking step: 10% serum for 1 hour at 25°C.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (ab183685)Verified customer
Formaldehyde-fixed, 3% hydrogen peroxide permeabilized B16F10 (melanoma) syngeneic mouse tumor tissue stained for CD4 with ab183685 (36mins, 37°C at a 1/1000 dilution) in immunohistochemical analysis. A Goat anti Rabbit IgG polyclonal HRP conjugate was used as the secondary.
Heat mediated antigen retrieval buffer/enzyme used: Tris/EDTA.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (ab183685)Zohar Gavish
Formaldehyde-fixed, non-permeabilized mouse spleen tissue stained for CD4 with ab183685 (30mins at a 1/1000 dilution) in immunohistochemical analysis. A Goat polyclonal HRP conjugate was used as the secondary.
Heat mediated antigen retrieval buffer/enzyme used: Leica ER2 solution.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (ab183685)Verified customer
Formaldehyde-fixed mouse colon, chronic mucosal inflammation tissue stained for CD4 with ab183685 (90 mins at a 1/1000 dilution) in immunohistochemical analysis. A Goat polyclonal biotin conjugate was used as the secondary.
Heat mediated antigen retrieval buffer/enzyme used: High pH CC1.
Blocking step: 1% BSA for 40 mins at 24°C.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (ab183685)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.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (ab183685)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.Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (ab183685)This image is courtesy of an anonymous Abreview.
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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (ab183685)
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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (ab183685)This image is courtesy of an Abreview submitted by Mr. Carl Hobbs.
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.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD4 antibody [EPR19514] (ab183685)This image is courtesy of an anonymous Abreview.
Formaldehyde-fixed, paraffin-embedded mouse thymus tissue stained for CD4 using ab183685 at 1/500 dilution in immunohistochemical analysis.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (92)
ab183685 has been referenced in 92 publications.
- Boucher AA et al. Cell type-specific mechanisms coupling protease-activated receptor-1 to infectious colitis pathogenesis. J Thromb Haemost 18:91-103 (2020). PubMed: 31539206
- Zhang Z et al. A Serum Metabolomic Study on Rats Induced by Polygoni Multiflori Radix and Polygoni Multiflori Radix Preparata by Pattern Recognition and Pathways Analysis. Biol Pharm Bull 43:306-318 (2020). PubMed: 31761827
- Lentscher AJ et al. Chikungunya virus replication in skeletal muscle cells is required for disease development. J Clin Invest 130:1466-1478 (2020). PubMed: 31794434
- Sun T et al. Combination Immunotherapy with Cytotoxic T-Lymphocyte-Associated Antigen-4 and Programmed Death Protein-1 Inhibitors Prevents Postoperative Breast Tumor Recurrence and Metastasis. Mol Cancer Ther 19:802-811 (2020). PubMed: 31796506
- Orgaz JL et al. Myosin II Reactivation and Cytoskeletal Remodeling as a Hallmark and a Vulnerability in Melanoma Therapy Resistance. Cancer Cell 37:85-103.e9 (2020). PubMed: 31935375