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Anti cd4

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Anti-CD4 is a laboratory reagent used for the detection and identification of CD4+ T cells. It is a monoclonal antibody that specifically binds to the CD4 antigen present on the surface of certain T lymphocytes. This reagent can be used in various immunological techniques, such as flow cytometry, to analyze the presence and proportion of CD4+ T cells in a sample.

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Lab products found in correlation

Anti cd8, by Agilent Technologies (6 mentions) Anti cd3, by Agilent Technologies (5 mentions) Anti granzymeb, by Agilent Technologies (2 mentions) Anti foxp3, by Abcam (2 mentions) Anti atg5, by Novus Biologicals (1 mentions) Anti p smad2 3, by Santa Cruz Biotechnology (1 mentions) Envision flex target retrieval solution, by Agilent Technologies (1 mentions) Envision flex, by Agilent Technologies (1 mentions) Anti nkp46, by R&D Systems (1 mentions) Anti ki67, by Agilent Technologies (1 mentions) Anti cd40 1c10, by Thermo Fisher Scientific (1 mentions) Vector novared substrate kit, by Vector Laboratories (1 mentions) Biotinylated anti rat igg, by Jackson ImmunoResearch (1 mentions) Hematoxylin, by Merck Group (1 mentions) Hematoxylin, by Leica (1 mentions) Anti ccl3, by Thermo Fisher Scientific (1 mentions) Fetal calf serum (fcs), by Harvard Bioscience (1 mentions) Interleukin 2 (il 2), by Roche (1 mentions) Interleukin 4 (il 4), by R&D Systems (1 mentions) Rpmi 1640 medium, by Merck Group (1 mentions)

Spelling variants of this product

Mouse anti-CD4 (2 citations) Anti-CD4 APC (2 citations) Anti-CD3 and anti-CD4 (2 citations) Anti-TM (1 citations)

12 protocols using anti cd4

1

Immunofluorescence Analysis of LC3, Atg5, CD4, CD8

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IIF analysis was performed on paraffin-embedded sections in order to characterize LC3-II-expressing cells and to assess whether LC3-II colocalized with Atg5, CD4, and CD8 cells. Sections were incubated with anti-human-LC3 (Novus Biologicals, Littleton, CO, USA), anti-Atg5 (Novus Biologicals), anti-CD3, anti-CD4, and anti-CD8 Abs (Dako) and then labeled with FITC- or Rhodamine Red-conjugated anti-mouse or anti-rabbit Abs plus RNasi (200 ng/mL) and counterstained using Toto-3 iodide (642/660; Invitrogen, Monza MB, Italy). Confocal analysis was used to acquire fluorescence staining.
Alessandri C., Ciccia F., Priori R., Astorri E., Guggino G., Alessandro R., Rizzo A., Conti F., Minniti A., Barbati C., Vomero M., Pendolino M., Finucci A., Ortona E., Colasanti T., Pierdominici M., Malorni W., Triolo G, & Valesini G. (2017). CD4 T lymphocyte autophagy is upregulated in the salivary glands of primary Sjögren’s syndrome patients and correlates with focus score and disease activity. Arthritis Research & Therapy, 19, 178.
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2

Immunohistochemical Profiling of Immune Cells

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Immunohistochemical staining was done at the Mayo Pathology Resource Core Facility. Antibodies used were purified anti CD8 (DAKO), purified anti CD4 (DAKO), purified anti FoxP3 (Abcam), purified anti granzymeB (DAKO), purified anti IL-15R (Biorbyt), and purified anti psmad2/3 (Santa Cruz Biotechnology).
Marietta E.V., Nadeau A.M., Cartee A.K., Singh I., Rishi A., Choung R.S., Wu T.T., Rubio-Tapia A, & Murray J.A. (2015). Original Scientific Paper: Immunopathogenesis of Olmesartan-Associated Enteropathy. Alimentary pharmacology & therapeutics, 42(0), 1303-1314.
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3

Immunohistochemical Analysis of Tumor-Infiltrating Immune Cells

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Paraffin-embedded tissue blocks from patient tumours prepared for routine clinical histopathology were obtained from Sahlgrenska University Hospital. Sections (3–4  μm) from paraffin blocks were placed on glass slides and treated in Dako PT-Link using EnVision™ FLEX Target Retrieval Solution (high pH). The following primary antibodies were used: anti-CD3 (Dako/Agilent Technologies, Santa Clara, CA, USA; IR503), anti-CD4 (Dako; 4B12), anti-CD8 (Dako; C8/144B), anti-NKp46 (R&D Systems, Minneapolis, MN, USA; 195314), anti-granzyme B (Novocastra/Leica Biosystems, Wetzlar, Germany; 11F1), anti-PD-L1 kit (Dako; PD-L1 IHC 28-8 pharmDx; according to manufacturer’s instructions), and anti-Ki-67 (Dako; MIB-1). Immunohistochemical staining was performed in a Dako Autostainer Link using EnVision™ FLEX according to the manufacturer’s instructions (DakoCytomation). EnVision™ FLEX+ (LINKER) mouse was used for anti-NKp46, anti-CD25, anti-granzyme B, and anti-Ki-67. Positive and negative controls were included in each run. Each staining was evaluated by a board-certified pathologist (O.N.). The Ki67 index was calculated by manually counting the percentage of labelled tumour cell nuclei on printed images [28 (link)].
Hofving T., Liang F., Karlsson J., Yrlid U., Nilsson J.A., Nilsson O, & Nilsson L.M. (2021). The Microenvironment of Small Intestinal Neuroendocrine Tumours Contains Lymphocytes Capable of Recognition and Activation after Expansion. Cancers, 13(17), 4305.
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4

Immunohistochemical Characterization of Thyroid Tissue

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Frozen or formaldehyde fixed paraffin sections of thyroids were blocked
with 5% BSA in PBS, and endogenous peroxidase was inhibited by
incubation with 0.3% H2O2 for 30 min. Anti-TTF-1 (H-190, Santa Cruz
Biotechnology), anti-CD40 (1C10; eBioscience for frozen sections or C-20 (sc975;
Santa Cruz for paraffin sections), anti-human CD3 (rabbit polyclonal, Dako),
anti-CD4 (clone GK1.5, supernatant) or anti-CD8 (clone 53.6, supernatant) were
used as primary Ab. For staining of frozen sections (CD4, CD8 and CD40),
biotinylated anti-rat IgG (Jackson ImmunoResearch Laboratories Inc., West Grove,
PA) was used as secondary Ab (1:500), followed by avidin-HRP binding using a
Vectastain Elite PK-6100 kit (Vector Laboratories, Burlingame, CA). Peroxidase
activity was visualized using a Vector Nova-Red Substrate Kit (Vector). TTF1,
CD3 and CD40 staining of paraffin sections was done by IDEXX RADIL, Columbia,
MO. They were developed with biotinylated anti-rabbit or anti-goat IgG at
previously determined optimal concentrations followed by avidin-HRP and
visualized using diaminobenzidine tetrahydrochloride (DAB) as the chromogen.
Kayes T.D., Weisman G.A., Camden J.M., Woods L.T., Bredehoeft C., Downey E.F., Cole J, & Braley-Mullen H. (2016). A new murine model of early onset autoimmune thyroid disease/hypothyroidism and autoimmune exocrinopathy of the salivary gland. Journal of immunology (Baltimore, Md. : 1950), 197(6), 2119-2130.
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5

Immunohistochemical Analysis of Skin Biopsies

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Cryostat frozen skin biopsies were sliced into 10-μm thick sections and fixed in acetone or 10% neutral buffered formalin. Sections were incubated with anti-CD4 (1:10; Dako) and anti-CCL3 (1:20; Thermo Fisher) antibodies overnight at 4 °C. Slides were counterstained with hematoxylin (Sigma-Aldrich), dehydrated, and mounted. After counterstaining with hematoxylin, sections were examined using a Leica microscope and analyses performed on two independent fields per section.
Eoli M., Corbetta C., Anghileri E., Di Ianni N., Milani M., Cuccarini V., Musio S., Paterra R., Frigerio S., Nava S., Lisini D., Pessina S., Maddaloni L., Lombardi R., Tardini M., Ferroli P., DiMeco F., Bruzzone M.G., Antozzi C., Pollo B., Finocchiaro G, & Pellegatta S. (2019). Expansion of effector and memory T cells is associated with increased survival in recurrent glioblastomas treated with dendritic cell immunotherapy. Neuro-oncology Advances, 1(1), vdz022.
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6

Lymphocyte Isolation and Cytokine Assays

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All tissue culture media contained RPMI-1640 medium (Sigma Aldrich, St Louis, MO, USA), 10% heat-inactivated FCS (Biochrom AG, Berlin, Germany), and 100 U/mL of penicillin and 100 μg/mL of streptomycin (Biochrom). Separating of lymphocytes was conducted using Ficoll–Hypaque (Seromed, Biochrom KG, Berlin, Germany). IL-2 was purchased from Roche Diagnostics GmbH (Mannheim, Germany), IL-4 was purchased from R&D Systems, Inc., (Minneapolis, MN, USA), and recombinant human Interleukin-12 (RH) IL-12 was purchased from Gibco® Invitrogen Corporartion (Camarillo, CA, USA). ELISA (Bender MedSystems GmbH) was used for the IFN-γ assays. Monoclonal antibodies anti-CD3, anti-CD4 and anti-CD8 obtained from DakoCytomation (Ely, Cambridgeshire, UK); anti-CD25 purchased from Fitzgerald Industries International (Acton, MA, USA) were used for the flow cytometric analysis.
Gabeen A.A., Abdel-Hamid F.F., El-Houseini M.E, & Fathy S.A. (2014). Potential immunotherapeutic role of interleukin-2 and interleukin-12 combination in patients with hepatocellular carcinoma. Journal of Hepatocellular Carcinoma, 1, 55-63.
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7

Immunohistochemical Analysis of Inflammatory Markers

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The tissue sections were processed with an Envision Flex+kit (DAKO), blocking endogenous peroxidase activity for 5 min, and then incubated with primary antibodies. The reaction was visualized by Envision Flex + horseradish peroxidase for 20 min and then with diaminobenzidine for 10 min. Sections were counterstained with Mayer’s hematoxylin for 5 min. Primary antibodies used for the study were: anti-CD4 (Dako, Santa Clara, CA, USA, ready to use, or Abcam (Cambridge, UK, ab846), 1:50 for 20 min), anti-IBA1 (Wako, Osaka, Japan, 1:300 for 30 min), anti-GFAP (Dako, ready to use, for 20 min), anti-TNFa (ab6671 from Abcam, Cambridge, UK, 1:2000 for 45 min), anti-CCl2 (66272, Proteintech, Rosemont, IL, USA, 1:1000), anti-CCl20 (ab9829, Abcam; 1:200 overnight), anti-CX3CL1 (14-7986 from Invitrogen, Waltham, MA, USA, 1:200 overnight).
Balzano T., Leone P., Ivaylova G., Castro M.C., Reyes L., Ramón C., Malaguarnera M., Llansola M, & Felipo V. (2021). Rifaximin Prevents T-Lymphocytes and Macrophages Infiltration in Cerebellum and Restores Motor Incoordination in Rats with Mild Liver Damage. Biomedicines, 9(8), 1002.
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8

Immunohistochemical Analysis of cSCC

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Ethical approval for the study was provided by the South Central Hampshire B NRES Committee (reference number 07/H0504/187). Archived formalin-fixed paraffin-embedded (FFPE) cSCC samples were obtained from University Hospital Southampton NHS Foundation Trust. Immunohistochemistry was performed as described previously (21 (link)), with microwave antigen retrieval performed in either 10 μM citrate or 1.6 μM EDTA buffers. Primary antibodies included anti-CD3 (1:200, DAKO), anti-CD4 (1:100, DAKO), anti-CD8 (1:20, Abcam), anti-CD25 (1:50, Novocastra), anti-FOXP3 (1:50, Abcam) and anti-OX40 (1:50, BD Biosciences), and biotinylated goat anti-mouse (1:200, DAKO) or swine anti-rabbit (1:400, DAKO) were used as secondary antibodies. For cell quantification, 5 representative images at 40× magnification were taken from each immunostained cSCC section and analysed using ImageJ software.
Lai C., August S., Albibas A., Behar R., Cho S.Y., Polak M.E., Theaker J., MacLeod A.S., French R.R., Glennie M.J., Al-Shamkhani A, & Healy E. (2016). OX40+ regulatory T cells in cutaneous squamous cell carcinoma suppress effector T cell responses and associate with metastatic potential. Clinical cancer research : an official journal of the American Association for Cancer Research, 22(16), 4236-4248.
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9

Kidney Allograft Biopsy Analysis

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Kidney allograft biopsies were obtained per protocol time point or suspected rejection. 36 biopsies were dual stained for anti-CD57 (DAKO, Carpinteria, CA) and anti-CD4 (DAKO, Carpinteria, CA). Blinded analysis was performed to determine cell density and localization.
Espinosa J., Herr F., Tharp G., Bosinger S., Song M., Farris AB I.I.I., George R., Cheeseman J., Stempora L., Townsend R., Durrbach A, & Kirk A.D. (2016). CD57+ CD4 T cells underlie belatacept-resistant allograft rejection. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 16(4), 1102-1112.
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10

Immunohistochemical Profiling of Tissue Samples

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For immunohistochemistry (IHC) staining, the following antibodies were applied on formalin-fixed, paraffin-embedded tissue using a routine immunoperoxidase technique [15 , 16 ] and experimental sequential IHC [17 (link)]: anti-CD3 (Dako M7254), anti-CD4 (Dako M7319), anti-CD8 (Dako M7103), anti-CD20 (Dako M0755), anti-Bcl2 (DakoM0887), anti-MPO (Dako C7246), anti-CD61 (Dako C7280). Two chromogens were used, DAB (DAB chromogen) as fix and NovaRed (VECTOR NovaRed) as removable chromogen for sequential IHC [17 (link)].
Weis C.A., Grießmann B.W., Scharff C., Detzner C., Pfister E., Marx A, & Zoellner F.G. (2015). On the representation of cells in bone marrow pathology by a scalar field: propagation through serial sections, co-localization and spatial interaction analysis. Diagnostic Pathology, 10, 151.
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