Anti human cd3

Manufactured by Agilent Technologies

Sourced in Denmark

Anti-human CD3 is a laboratory equipment used for the detection and quantification of CD3, a protein complex found on the surface of T cells. It is a valuable tool for immunological research and analysis.

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Lab products found in correlation

Discovery xt platform, by Roche (1 mentions) Anti human cd8 clone c8 144b, by Agilent Technologies (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Alexa fluor 488 goat anti mouse igg, by Thermo Fisher Scientific (1 mentions) Cx41 light microscope, by Olympus (1 mentions) Anti human cd20, by Thermo Fisher Scientific (1 mentions) Hematoxylin, by Merck Group (1 mentions) Proteinase k, by Agilent Technologies (1 mentions) Harris hematoxylin solution, by Merck Group (1 mentions) Vectastain elite abc kit, by Vector Laboratories (1 mentions) Anti human cd8, by Abcam (1 mentions) Anti gfap, by Abcam (1 mentions) Prolong gold antifade mountant with dapi, by Thermo Fisher Scientific (1 mentions) Anti p24, by Abcam (1 mentions) Anti hu nuclei, by Merck Group (1 mentions)

Close spelling variants of this product

Anti-CD3 (82 citations) Polyclonal rabbit anti-human CD3 (14 citations) Rabbit anti-human CD3 (11 citations) Mouse anti-human CD3 (8 citations) Anti-human CD3 antibody (4 citations) Rabbit anti-human CD3 antibody (4 citations) Monoclonal mouse anti-human CD3 (clone F7.2.38; (3 citations) Polyclonal rabbit anti-human CD3 antibodies (2 citations) Anti-human CD3 (clone F7.2.38) (2 citations) Mouse anti-human CD3 antibody (2 citations)

7 protocols using anti human cd3

Histological Assessment of Kidney Injury

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The formalin-fixed tissue was paraffin embedded, cut in 4μm
sections, mounted and stained with Gomori’s one-step Trichrome. Tubular
protein casts and glomeruli injury were quantified as previously
described^{11 (link)}. Glomeruli
were scored by a blinded observer on a scale from 0–4. Two distinct
populations were scored, cortical and juxtamedullary glomeruli, determined by
their depth in the kidney. Separate sections were stained with CD3+
(Anti-human CD3; 1:100; DAKO) for qualitative assessment of T-cell
localization.

Evans L.C., Petrova G., Kurth T., Yang C., Bukowy J.D., Mattson D.L, & Cowley AW J.r. (2017). INCREASED PERFUSION PRESSURE DRIVES RENAL T-CELL INFILTRATION IN THE DAHL SALT-SENSITIVE RAT. Hypertension (Dallas, Tex. : 1979), 70(3), 543-551.

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Quantifying Immune Cell Populations in FFPE Tissue

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FFPE specimens were sectioned at five-micrometers and stained on the DISCOVERY XT platform (Ventana Medical Systems, Inc., Tucson, U.S.A). TAM and microglia were identified using antibodies against IBA1. Antibodies targeting CD3 and CD8 stained the entire T cell population and cytotoxic T cells, respectively. Regulatory T cells were stained by antibodies against forkhead box P3 (FOXP3). Due to the lack of a validated antibody, the CD4⁺ T cell number was estimated to be the difference between the CD3⁺ and CD8⁺ T cell numbers according to published protocol.³⁵ In six cases, the number of CD8⁺ T cells exceeded or was identical to the number of CD3⁺ T cells, so that the CD4⁺ T cell population was considered to be equal to the FOXP3⁺ T cell number, since the latter constitutes a subpopulation of CD4⁺ T cells. The following primary antibodies were used: Anti-IBA1 (1:500, rabbit polyclonal, #019–19741, Wako Pure Chemical Ind., Ltd., Osaka, Japan); anti-human FOXP3, clone 259D (1:100, mouse monoclonal, #320202, BioLegend, San Diego, U.S.A); anti-human CD8, clone C8/144B (1:100, mouse monoclonal, code M7103, Dako, Glostrup, Denmark); anti-human CD3 (1:100, rabbit polyclonal, code A0452, Dako, Glostrup, Denmark).

Kaffes I., Szulzewsky F., Chen Z., Herting C.J., Gabanic B., Velázquez Vega J.E., Shelton J., Switchenko J.M., Ross J.L., McSwain L.F., Huse J.T., Westermark B., Nelander S., Forsberg-Nilsson K., Uhrbom L., Maturi N.P., Cimino P.J., Holland E.C., Kettenmann H., Brennan C.W., Brat D.J, & Hambardzumyan D. (2019). Human Mesenchymal glioblastomas are characterized by an increased immune cell presence compared to Proneural and Classical tumors. Oncoimmunology, 8(11), e1655360.

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Vα7.2 Immunodetection in Liver Samples

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Vα7.2 immunodetection was performed in frozen sections (10 μm) of normal (F0, n = 4) or subnormal (F1, n = 3) liver samples from patients who underwent resection surgeries for non-hepatocellular primary tumor (n = 3), colorectal (n = 2), or kidney (n = 1) cancer liver metastasis, and showed no (F0 n = 4) or mild (F1 n = 3) fibrosis and no alteration in liver biological tests. Liver samples from patients with cirrhosis were obtained from non-tumoral part of HCC resection (n = 3) or liver explant during liver transplantation (n = 2).
Tissue sections were incubated overnight at 4 °C with either anti-human CD3 (DAKO, 1:50 in PBS containing 1% BSA and 0.2% Triton X-100) or anti-human α-SMA (BioLegend, 1:200). Slides were then incubated for 1 h at room temperature with secondary antibody-HRP working solution, followed by Opal 570 fluorophore. Incubation with purified anti-human Vα7.2 antibody (BioLegend, 1:50) was performed overnight at 4 °C, followed by incubation in Alexa Fluor 488 goat anti-mouse IgG (1:200, Life Technologies) for 1 h at room temperature. Nuclear counterstaining was obtained with DAPI (1:10,000, Thermo). Images were acquired by confocal microscopy (Confocal Zeiss LSM 780). Vα7.2-positive cells were assessed (0: absent; +: positive) and their location was determined (sinusoidal space and mesenchymal space).

Hegde P., Weiss E., Paradis V., Wan J., Mabire M., Sukriti S., Rautou P.E., Albuquerque M., Picq O., Gupta A.C., Ferrere G., Gilgenkrantz H., Kiaf B., Toubal A., Beaudoin L., Lettéron P., Moreau R., Lehuen A, & Lotersztajn S. (2018). Mucosal-associated invariant T cells are a profibrogenic immune cell population in the liver. Nature Communications, 9, 2146.

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Immunohistochemical Analysis of Humanized NSG Mice Spleen

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All of the humanized NSG mice spleen tissues were harvested, fixed in 10% formalin and embedded in paraffin. Immunohistochemistry was performed on 4 μm tissue sections fixed on slide glass, deparaffinized with xylene and rehydrated with ethanol. Immunohistochemistry (IHC) was performed using Dako Real Envision Detection System, Peroxidase/DAB Rabbit/Mouse (DAKO, Glostrup, Denmark) kit according to the manufacturer’s instructions. Primary antibody used for IHC was rabbit polyclonal anti-human CD3 (1:1000, DAKO, Glostrup, Denmark). Stained slides were observed using an Olympus CX41 light microscope (Olympus, Japan) with 10X/22 numeric aperture and 40x/0.75 numeric aperture objective, and photographs were taken by a microscope digital camera DP50 (Olympus, Japan) and image-pro plus 5.1 Software. All samples were counterstained with hematoxylin and eosin (H&E).

Kwon D.H., Park J.B., Lee J.S., Kim S.J., Choi B, & Lee K.Y. (2021). Human delta like 1-expressing human mesenchymal stromal cells promote human T cell development and antigen-specific response in humanized NOD/SCID/IL-2Rnull (NSG) mice. Scientific Reports, 11, 10603.

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Immunohistochemical Analysis of Tumor-Infiltrating Immune Cells

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The presence of tumoral cells and infiltrating T and B cells in the biopsies was determined by hematoxylin and eosin (HE) and immunohistochemistry (IHC). Antibodies used for the paraffin-embedded sections of the TNBC biopsy were anti-human CD3 (clone 2GV6), anti-human CD4 (clone SP35), anti-human CD8 (clone SP57), and anti-human CD20 (clone L26) (Ventana Medical System, Inc.). IHC of the XABCL tissue was performed in samples fixed in 10% neutral buffered formalin. Transverse sections were used for processing. Morphological evaluation was performed on 5-µm paraffin-embedded sections, stained with HE. The primary antibodies used were rabbit polyclonal anti-human CD3 (Dako) and rabbit polyclonal anti-human CD20 (Thermo Fisher). Sections were incubated with a labeled polymer [anti-rabbit for CD3 and CD20 (Agilent-Dako)] according to the instructions of the manufacturer. IHC was completed using 3,3′-diaminobenzidine (DAB) and counterstaining in hematoxylin. Positive controls of the XABCL IHC were performed using normal mouse spleen and lymph node. In all the experiments, for the negative control, an isotype-specific immunoglobulin was used to substitute the primary antibody as negative control; no immunostaining was detected in these sections.

Aran A., Peg V., Rabanal R.M., Bernadó C., Zamora E., Molina E., Arribas Y.A., Arribas J., Pérez J., Roura-Mir C., Carrascal M., Cortés J, & Martí M. (2021). Epstein–Barr Virus+ B Cells in Breast Cancer Immune Response: A Case Report. Frontiers in Immunology, 12, 761798.

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Immunohistochemical Analysis of Skin Biopsies

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Skin and subcutaneous tissue biopsies were fixed in 10% phosphate-buffered formalin for 24 hours and paraffin embedded. The samples were sectioned in 5-µm-thick slides for hematoxylin and eosin (H&E) and immunohistochemical staining. Specifically, the paraffin samples were cleared and rehydrated through a series of xylene and ethanol and stained with hematoxylin (Sigma-Aldrich, St. Louis, Mo.) and eosin (Thermo Fisher Scientific, Cheshire, UK). Presence of CD3, CD4, CD8, and CD20 was evaluated using VECTASTAIN Elite ABC Kit (Vector Laboratories, Burlingame, Calif.) for anti-human CD3 (1:1000, Agilent), anti-human CD4 (1:200, Agilent), anti-human CD8 (1:500, Abcam) and anti-human CD20 (1:500, Agilent). Briefly, proteinase K (Dako, Carpinteria, Calif.) was utilized for antigen retrieval and nonspecific binding was blocked using normal horse serum. For isotype control, mouse IgG was utilized at the same dilution as that of the primary antibody. Slides were incubated with biotinylated secondary antibody for 30 minutes followed by 30-minute incubation with RTU VECTASTATIN ABC Reagent. Then 30 µl of ImmPACT DAB Reagent was diluted in 1 mL of ImmPACT DAB Dilutent (Vector Laboratories, Burlingame, Calif.) and applied to the slides. They were counterstained with Harris hematoxylin solution (Sigma-Aldrich, St. Louis, Mo.) and dehydrated and cleared in ethanol and xylene, respectively.

Haykal S., Barone N., Rostami S., Moshkelgosha S., Juvet S., Keshavjee S, & Ghazarian D. (2023). Immunohistochemical Analysis of Lymphocyte Populations in Acute Skin Rejection: The University Health Network Addition to the Banff Classification. Plastic and Reconstructive Surgery Global Open, 11(3), e4831.

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Immunohistochemical Analysis of Mouse Brain

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Mouse brains were fixed in 4% paraformaldehyde for 24 hours at 4 ºC then kept in 30% sucrose solution at 4 ºC. Forty μm thick coronal sections were used in these studies. Tissue sections were incubated in primary antibodies overnight treatment at room temperature. Primary antibodies are as follows: Anti-huGFAP (1:200–500, BioLegend, San Diego, CA, 837202), anti-GFAP (1:1000, Abcam, Cambridge, UK, ab4674), anti-huNuclei (1:200, Millipore, MAB1281), anti-human CD3 (1:200, Agilent, Santa Clara, CA, A0452) and anti-p24 (Abcam, ab53841). After washing sections of primary antibody they were incubated in secondary antibody (1:200, Life Technologies) for 1 h at room temperature. After washing the sections were covered with ProLong Gold Antifade Mountant with DAPI (Thermo Fisher).

Lutgen V., Narasipura S.D., Barbian H.J., Richards M., Wallace J., Razmpour R., Buzhdygan T., Ramirez S.H., Prevedel L., Eugenin E.A, & Al-Harthi L. (2020). HIV infects astrocytes in vivo and egresses from the brain to the periphery. PLoS Pathogens, 16(6), e1008381.

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