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Anti cd4

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Anti-CD4 is a monoclonal antibody that binds to the CD4 cell surface glycoprotein, which is primarily expressed on the surface of T helper cells. The core function of Anti-CD4 is to assist in the identification and characterization of CD4+ cells in various research and analytical applications.

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Lab products found in correlation

Anti cd8, by Santa Cruz Biotechnology (5 mentions) Dm6 microscope, by Leica (3 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Anti il 17, by Santa Cruz Biotechnology (2 mentions) Goat anti rabbit igg, by Vector Laboratories (1 mentions) Anti nitrotyrosine, by Merck Group (1 mentions) Avidin biotin peroxidase complex, by Vector Laboratories (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Fluorescein isothiocyanate (fitc), by Santa Cruz Biotechnology (1 mentions) Lsm 700 confocal microscope, by Zeiss (1 mentions) Anti il 17, by Abcam (1 mentions) Anti α4β7, by BioLegend (1 mentions) Biotinylated anti rabbit igg, by Vector Laboratories (1 mentions) Dab peroxidase substrate kit, by Vector Laboratories (1 mentions) Biotinylated anti goat igg, by Vector Laboratories (1 mentions) Anti il 16, by Santa Cruz Biotechnology (1 mentions) Streptavidin peroxidase complex, by Thermo Fisher Scientific (1 mentions) Dfc425, by Leica (1 mentions) Anti cd68, by Santa Cruz Biotechnology (1 mentions) Biotin conjugated goat anti rabbit immunoglobulin g, by Vector Laboratories (1 mentions)

Close spelling variants of this product

Mouse anti-CD4 (3 citations) Anti-CD4+ (sc-13573 L (2 citations)

18 protocols using anti cd4

1

Immunohistochemical Analysis of Inflammatory Markers

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Immunohistochemical examination was performed as previously described [31 (link)], 4 days after DNBS administration. The sections were incubated overnight with the following primary antibodies: anti-CD4+ (Santa Cruz Biotechnology (SCB), 1:100 in PBS, v/v, sc-19641, D.B.A, Milan, Italy); anti-CD8+ (SCB, 1:100 in PBS, v/v, sc-1177, D.B.A, Milan, Italy); anti-ICAM-1 (SCB, 1:100 in PBS, v/v, sc-107, D.B.A, Milan, Italy), anti-P-selectin (1:100 in PBS, v/v, sc-8419 SCB, D.B.A, Milan, Italy), anti-PARP (1:100 in PBS, v/v, sc-8007 SCB, D.B.A, Milan, Italy), and anti-nitrotyrosine (1:200 in PBS, v/v, Millipore, D.B.A, Milan, Italy). All slices were washed with PBS and then treated as previously reported [32 (link)]. Briefly, slices were incubated with universal secondary antibody. Specific labeling was detected with a biotin- conjugated goat anti-rabbit IgG and avidin-biotin peroxidase complex (Vector).
Five stained sections from each mouse were scored in a blinded fashion and observed using a Leica DM6 microscope (Leica Microsystems SpA, Milan, Italy) following a typical procedure [33 (link)]. The histogram profile is related to the positive pixel intensity value obtained.
Impellizzeri D., Fusco R., Genovese T., Cordaro M., D’Amico R., Trovato Salinaro A., Ontario M.L., Modafferi S., Cuzzocrea S., Di Paola R., Calabrese V, & Siracusa R. (2022). Coriolus Versicolor Downregulates TLR4/NF-κB Signaling Cascade in Dinitrobenzenesulfonic Acid-Treated Mice: A Possible Mechanism for the Anti-Colitis Effect. Antioxidants, 11(2), 406.
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2

Visualizing Intestinal Immune Cells

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Small intestine tissues were stained with anti-CD4 (Santa Cruz Biotechnology, Dallas, TX, USA), anti-IL-17 (Abcam), and anti-α4β7 (Biolegend, San Diego, CA, USA) primary antibodies at 4°C overnight, followed by secondary antibodies conjugated with FITC (Santa Cruz Biotechnology, APC (Thermo Fisher Scientific, Rockford, IL, USA) and PE (Thermo Fisher Scientific) incubated at room temperature for 2 h. Nuclei were stained with 4,’6-diamidino-2-phenylindole (DAPI; Invitrogen, Carlsbad, CA). Confocal images were analyzed using an LSM 700 confocal microscope (Zeiss, Oberkochen, Germany) at 200× magnification.
Hwang S.H., Woo J.S., Moon J., Yang S., Park J.S., Lee J., Choi J., Lee K.H., Kwok S.K., Park S.H, & Cho M.L. (2021). IL-17 and CCR9+α4β7– Th17 Cells Promote Salivary Gland Inflammation, Dysfunction, and Cell Death in Sjögren’s Syndrome. Frontiers in Immunology, 12, 721453.
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3

Ovarian Expression of IL16 and CD4

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Immunohistochemical procedures were performed for IL16 and its known receptor CD4 in order to localize expression in the developing ovary. Formalin-fixed tissues from 14-day old and 19-day old rat pups were used for immunohistochemical staining as previously described (Nilsson et al., 2002 (link)). The ovary sections were incubated with a blocking buffer [10% goat serum (normal goat serum) or 10% rabbit serum (normal rabbit serum)], followed by the primary antibody, either anti- IL16 (Santa Cruz Biotechnology Inc., cat# sc-7902, USA) (made in rabbit, 0.2 μg/ml, 0.8 μg/ml, or 4 μg/ml) or anti CD4 (Santa Cruz Biotechnology Inc., cat# sc-1140, USA) (made in goat, 0.2 μg/ml, 0.8 μg/ml, or 4 μg/ml) for antigen binding. Secondary antibody was biotinylated anti-rabbit IgG made in goat (Vector Laboratories Inc., cat# BA-1000, USA) or biotinylated anti-goat IgG made in rabbit (Vector Laboratories Inc., cat# BA-5000, USA). The ovary sections were incubated with Streptavidin Peroxidase Complex (Invitrogen, cat# 50-420Z, USA) for enzyme coupling. A color reaction was performed using a DAB peroxidase substrate kit (Vector Laboratories Inc., cat# SK-4100, USA). Images were taken of the stained sections and the results were qualitatively analyzed.
Feeney A., Nilsson E, & Skinner M.K. (2014). Cytokine (IL16) and Tyrphostin Actions on Ovarian Primordial Follicle Development. Reproduction (Cambridge, England), 148(3), 321-331.
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4

Immunohistochemical Visualization of CD4

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The sections of the colons were incubated with anti-CD4 (1:200 dilution; Santa Cruz Biotechnology, Inc., Dallas, TX, USA) antibodies, followed by secondary antibodies conjugated with AlexaFluor 488 (1:1500 dilution) from Invitrogen. The immunostained antigens were visualized using a Leica DFC425 fluorescence microscope [Leica Microsystems (Schweiz) AG, Heerbrugg, Switzerland].
Shi Y., Liu Z., Cui X., Zhao Q, & Liu T. (2020). Intestinal vitamin D receptor knockout protects from oxazolone-induced colitis. Cell Death & Disease, 11(6), 461.
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5

Immunohistochemistry Analysis of Heart Tissue

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An immunohistochemical analysis were performed as described previously [40 (link),41 (link)]. The heart tissues were fixed in 10% buffered formaldehyde and 7 μm sections were prepared from paraffin-embedded tissues. After deparaffinization, endogenous peroxidase was quenched with 0.3% H2O2 in 60% methanol for 30 min. The sections were permeabilized with 0.1% Triton X-100 in phosphate-buffered saline (PBS) for 20 min. Non-specific adsorption was minimized by incubating the section in 2% normal goat serum in phosphate-buffered saline for 20 min. Endogenous biotin or avidin binding sites were blocked by sequential incubation for 15 min with avidin and biotin. The sections were probed with the following primary antibodies: anti-CD4 (Santa Cruz Biotechnologies, sc-13573, Dallas, TX, USA) and anti-CD68 (Santa Cruz Biotechnologies, sc-20060). The slides were then washed with PBS and incubated with a secondary antibody. Specific labeling was achieved with an avidin–biotin–peroxidase complex and biotin-conjugated goat anti-rabbit immunoglobulin G (Vector Lab, Milan, Italy) [42 (link)]. The stained sections were observed using a Leica DM6 microscope (Leica Microsystems SpA, Milan, Italy).
Interdonato L., Impellizzeri D., D’Amico R., Cordaro M., Siracusa R., D’Agostino M., Genovese T., Gugliandolo E., Crupi R., Fusco R., Cuzzocrea S, & Di Paola R. (2023). Modulation of TLR4/NFκB Pathways in Autoimmune Myocarditis. Antioxidants, 12(8), 1507.
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6

Quantitative analysis of tumor-infiltrating immune cells

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CD4+, CD8+ and Treg cells were measured by immunohistochemistry in tumor samples of M-406 bearing mice in the three tumor growth conditions. The histological sections of mammary tumors were deparaffinized, incubated overnight at 4°C with anti-CD4 (1:400, Santa Cruz Biotechnology, CA, USA), anti-CD8 (1:400, Santa Cruz Biotechnology) or anti-Foxp3 (1:50, eBioscience, MA, USA) and then, with the Vectastain Elite ABC kit (Vector Labs, USA). Sections were visualized with 3,3′-diaminobenzidine (Sigma, MO, USA) as chromogen. The number of positive cells was calculated in 30 fields at 1000×.
Giúdice A.D., Pagura L., Capitani M.C., Mainetti L.E., Scharovsky O.G., Di Masso R.J., Rico M.J, & Rozados V.R. (2020). Nonclassical roles for IFN-γ and IL-10 in a murine model of immunoedition. Future Science OA, 6(10), FSO589.
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7

Comprehensive Immunophenotyping of Murine Cells

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The following Abs and reagents were used in this study: FITC anti-mouse CD3ε (#100306), FITC anti-mouse CD19 (#152404), APC anti-mouse CD21/CD35 (#123412), PerCP/Cyanine5.5 anti-mouse CD23 (#101618), and Brilliant Violet 421 anti-mouse IgD (#405725) (all from BioLegend, CA, USA); PE anti-mouse CD45 (#12-0451-82) (Invitrogen, CA, USA); anti-TCR α/β (#sc-19600), anti-CD4 (#sc-19641), and anti-GAPDH (#sc-365062) (Santa Cruz Biotechnology, TX, USA); anti-MHC Class II (#ab180779), anti-ICOS (#ab175401), anti-ICOSL (#ab138354), anti-CD40 (#ab188181), and anti-CD40L (#ab2391) (Abcam, Cambridge, UK); and FITC anti-mouse CD169 (MOMA-1, #MCA947F) (Bio-Rad, CA, USA).
Chei S., Oh H.J., Lee K., Jin H., Lee J.Y, & Lee B.Y. (2020). Dysfunction of B Cell Leading to Failure of Immunoglobulin Response Is Ameliorated by Dietary Silk Peptide in 14-Month-Old C57BL/6 Mice. Frontiers in Nutrition, 7, 583186.
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8

Quantifying Regulatory T Cells in Lungs

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For these studies, bronchoalveolar cells (1 × 106/mL) were incubated with anti‐mouse CD3 FITC conjugate and B220 PE (R&D Systems, San Diego, California) for 30 min at 4°C. Cells were washed twice with cold PBS and resuspended in PBS; 10.000 events were acquired using a Becton Dickinson FACScan, and the data were analyzed using WinMIDI software. Cell populations were identified according to criteria described by Van Rijt and colleagues 25. To assess Treg cells were used antibodies anti‐FOXP3 (BD Biosciences Pharmingen), anti‐CD4 and anti‐CD25 (Santa Cruz Biotechnology). Cells from lymph nodes were fixed with paraformaldehyde 2% for 10 min at 37°C and permeabilized in ice with 90% methanol for 15 min. Before labeling, the cells were washed three times in HBSS and incubated with antibodies for 20 min. At the end of incubation, cells were washed and suspended in HBSS for further analyses on flow cytometer.
Mozzini Monteiro T., Ferrera Costa H., Carvalho Vieira G., Rodrigues Salgado P.R., da Silva Stiebbe Salvadori M.G., de Almeida R.N., de Fatima Vanderlei de Souza M., Neves Matias W., Andrade Braga V., Nalivaiko E, & Piuvezam M.R. (2016). Anti‐asthmatic and anxiolytic effects of Herissantia tiubae, a Brazilian medicinal plant. Immunity, Inflammation and Disease, 4(2), 201-212.
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9

Immunohistochemical Profiling of Cytokines and Immune Markers

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For the immunohistochemical evaluation, the following antibodies were used: anti-IL-4 (Santa Cruz Biotechnology, California, USA; 1:600), anti-IL-5 (Santa Cruz Biotechnology, California, USA; 1:100), anti-IL-10 (Santa Cruz Biotechnology, California, USA; 1:500), anti-IL-13 (Santa Cruz Biotechnology, California, USA; 1:700), anti-IL-17 (Santa Cruz Biotechnology, California, USA; 1:800), anti-CD4+ (Santa Cruz Biotechnology, California, USA; 1:25), anti-CD8+ (Santa Cruz Biotechnology, California, USA; 1:50), anti-MMP-9 (Santa Cruz Biotechnology, California, USA; 1:500) and anti-TIMP-1 (Santa Cruz Biotechnology, California, USA; 1:100).Immunohistochemistry was performed with the following sequence of procedures: antigenic recovery, endogenous peroxidase blockade and nonspecific binding blockade, incubation with the primary antibody, incubation with the secondary antibody and complex, counterstaining and assembly of the blades. The count was determined as described above for the evaluation of eosinophil density.
Florencio A.C., de Almeida R.S., Arantes-Costa F.M., Saraiva-Romanholo B.M., Duran A.F., Sasaki S.D., Martins M.A., Lopes F.D., Tibério I.F, & Leick E.A. (2019). Effects of the serine protease inhibitor rBmTI-A in an experimental mouse model of chronic allergic pulmonary inflammation. Scientific Reports, 9, 12624.
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10

Immunohistochemical Analysis of Tumor Markers

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Immunohistochemical localization was carried out as previously described by Scuderi et al. [6 (link)]. Tumor sections were incubated overnight at room temperature with the following primary antibodies: anti-Ki-67 (sc-23900, 1:100; Santa Cruz Biotechnology, Dallas, TX, USA), anti-IL-1β (sc-32294, 1:100; Santa Cruz Biotechnology, Dallas, TX, USA), anti-CD4 (sc-13573, 1:100; Santa Cruz Biotechnology, Dallas, TX, USA), anti-CD8 (sc-1177, 1:100; Santa Cruz Biotechnology, Dallas, TX, USA), and anti-CD30 (sc-19984, 1:100; Santa Cruz Biotechnology, Dallas, TX, USA). After this incubation, the sections were washed with PBS and incubated with a secondary antibody (Santa Cruz Biotechnology, Santa Cruz, CA, USA) for 1 h. The reaction was revealed by a chromogenic substrate (brown DAB), and counterstaining with Nuclear Fast Red. A negative control was performed using no primary antibody; specifically, tissue was incubated with the antibody diluent alone, followed by incubation with secondary antibodies and detection reagents. For immunohistochemistry, 20× (50 µm scale bar) and 40× (20 µm scale bar) magnification are shown.
Scuderi S.A., Casili G., Basilotta R., Lanza M., Filippone A., Raciti G., Puliafito I., Colarossi L., Esposito E, & Paterniti I. (2021). NLRP3 Inflammasome Inhibitor BAY-117082 Reduces Oral Squamous Cell Carcinoma Progression. International Journal of Molecular Sciences, 22(20), 11108.
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