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Cd3 antibody

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The CD3 antibody is a protein that binds to the CD3 complex, which is a group of proteins that are essential for the activation of T cells. The CD3 antibody is commonly used in research and clinical applications to identify and study T cells.

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Lab products found in correlation

Cd28 antibody, by Abcam (3 mentions) Immunomagnetic negative selection, by STEMCELL (1 mentions) Interleukin 2 (il 2), by Abcam (1 mentions) Transwell insert, by Corning (1 mentions) H 1500, by Vector Laboratories (1 mentions) Immunohistochemistry kit, by Wuhan Servicebio Technology (1 mentions) Cd8 antibody, by Cell Signaling Technology (1 mentions) Cd4 antibody, by Abcam (1 mentions) Foxp3 antibody, by Cell Signaling Technology (1 mentions) Setdb1 antibody, by Abcam (1 mentions) Alexa 488 conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) Fluorescence microscope, by Olympus (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions) Dhe staining solution, by Merck Group (1 mentions) Phospho stat3, by Cell Signaling Technology (1 mentions)

9 protocols using cd3 antibody

1

Transwell Assay for Cancer Cell Migration

Cited 2 times
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The process of this experiment was implemented as described in former study.18 (link),19 (link) Briefly, T cells were isolated from PBMC through immunomagnetic negative selection (STEMCELL Technologies, Canada). Isolated T cells were pre-activated by 100ng/mL CD3 antibody (Abcam, Shanghai, China) and recombination 10ng/mL IL-2 (Abcam, Shanghai, China). Cancer cells were transfected and cultured for 24h. The supernatant of cancer cell medium was collected. Polycarbonate membranes and cancer cell conditioned medium was placed in the bottom of the well. T cells (5×105) were seeded and cultured on Transwell inserts (diameter, 6.5 mm) containing 5- or 3-μm pore size (Costar, USA). After 2h of incubation, the Transwell inserts were lifted. The number of transitional cells was measured by a hemocytometer, cell migrated were normalized to NC group (cell migrated (%) = Cell migrated/Cell migrated in NC×100%). Each experiment was replicated triplicates.
Ye J., Chen X, & Lu W. (2021). Identification and Experimental Validation of Immune-Associate lncRNAs for Predicting Prognosis in Cervical Cancer. OncoTargets and therapy, 14, 4721-4734.
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2

Inflammatory Response and Src Kinase Analysis in cSCC

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cSCC tumor tissue was harvested from representative lesion prior to initiating treatment, at 2–3 weeks and at the end of treatment. Harvested tissues were fixed in 10% formalin for tissue processing. Histologic slides with hematoxylin and eosin staining were reviewed. The neutrophilic inflamatory response was assessed by morphology on H&E staining, inflammatory cells with a segmented nucleus and cytoplasmic granules. Murine CD3 positive T cells were detected using immunohistochemistry with an CD3 antibody, 1:100 (Abcam, Cambridge, MA). To assess cell numbers, five square millimeters of dermis was examined to determine the number of T cells and neutrophils in each biopsy specimen. Cells in the epidermis, hair follicle or fat layer were not counted. Biopsies from 3 different mice in the indicated cohorts were evaluated. Statistical significance was determined using a Student’s T test for the means.
Tissue sections were subjected to immunohistochemistry staining as previously reported (Zhao, 2009). Phosphorylated Src kinases was detected using pY416 rabbit Ab from Cell Signaling, #2101 at a 1:20 dilution. Total Src kinases were detected using the SRC2 (sc-18) antibody from Santa Cruz at a 1:100 diltion. All photomicrographs were obtained using a Keyence microscope.
Yang X., Daifallah A.E., Shankar S., Beer J., Marshall C., Dentchev T., Seykora F., D’Armas S., Hahn J., Lee V., Sabry H.H., Farag A.M, & Seykora J.T. (2019). Topical kinase inhibitors induce regression of cutaneous squamous cell carcinoma. Experimental dermatology, 28(5), 609-613.
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3

PBMC Activation and Co-Culture Assay

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Human peripheral blood mononuclear cells (PBMCs) were procured from the Cell Bank of the Chinese Academy of Sciences (Shanghai, China). PBMCs were activated using CD3 antibody (Abcam, Cambridge, UK) and CD28 antibody (Abcam). Activated PBMCs were then co-cultured for 72 h at a 5:1 ratio with Huh-7 cells and SMMC-7721 cells from different treatment groups (control group without treatment, IFN-γ with/without AS-IV treatment group). Huh-7 and SMMC-7721 cells were isolated and collected.
Ma Y., Li Y., Wu T., Li Y, & Wang Q. (2023). Astragaloside IV Attenuates Programmed Death-Ligand 1-Mediated Immunosuppression during Liver Cancer Development via the miR-135b-5p/CNDP1 Axis. Cancers, 15(20), 5048.
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4

Immunohistochemical Analysis of Npc1 and App Genotypes

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Mice brains of the indicated ages of Npc1+/+/App+/+, Npc1+/+/App−/−, Npc1−/−/App+/+, and Npc1−/−/App−/− genotypes were processed for immunohistochemistry as described by us [6 (link)]. Sections 25 μm thick were cut sagittally through the cerebellum and mounted onto gelatin-chrome alum-coated Superfrost microscope slides (VWR, Denver, USA). Slides were placed on a warming surface at 37 °C for 30 min and rinsed with PBS for 10 min six times. Slides were incubated in blocking solution (PBS with 5% normal goat serum, 1% bovine serum albumin and 0.2% of 10% Triton x100) for 2 h at room temperature. This step was followed by a 4 °C overnight incubation with CD3 antibody at 1:200 (Abcam 135372); incubation buffer consisted of PBS with 2% normal goat serum, 1% bovine serum albumin, and 0.1% Triton X-100. Following 3 washes in PBS with 0.1% Tween-20, slides were incubated in the dark with donkey anti-rabbit 488 secondary antibody (Abcam 21206) for 2 h at room temperature; incubation buffer consisted of PBS with 2% normal goat serum, 1% bovine serum albumin and 0.1% Triton X-100. Samples were washed twice in PBS with 0.1% Tween-20 and once with PBS. Slides were mounted in Vectashield/DAPI hard-set mounting medium (Vectashield H-1500).
Shin S.D., Shin A., Mayagoitia K., Siebold L., Rubini M., Wilson C.G., Bellinger D.L, & Soriano S. (2019). Loss of amyloid precursor protein exacerbates early inflammation in Niemann-Pick disease type C. Journal of Neuroinflammation, 16, 269.
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5

Immunohistochemical Analysis of Immune Cell Markers

Cited 1 time
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Tissues were fixed overnight in 4% paraformaldehyde and embedded in paraffin. All IHC staining was performed on 4-µm sections. After deparaffinization, rehydration, antigen unmasking, and endogenous peroxidase blocking, sections were blocked in 5% BSA with 0.1% Triton X-100 in PBS. Sections were incubated overnight at 4°C with primary antibodies. The details of antibodies are as follows: CD3 antibody (Abcam; cat #ab5690; 1:200); CD4 antibody (Abcam; cat #ab183685; 1:200); CD8 antibody (Cell Signaling Technology; cat #98941; 1:200); Foxp3 antibody (Cell Signaling Technology; cat #12653; 1:200); SETDB1 antibody (Abcam; cat #ab12317; 1:200); and FOXM1 antibody (Abcam; cat #ab232649; 1:200). Tissue sections were then stained using an Immunohistochemistry kit (Servicebio; cat #G1215; cat #G1215-200T; cat #G1216-200T). Tissues were observed under a microscope and photographed after being counterstained with hematoxylin, dried, and mounted. For assessment of CD3+, CD4+, CD8+, and Foxp3+ T cell infiltration, positively stained cells were counted from five high-power random fields of each sample, and the numbers averaged for each field. The IHC scores for SETDB1 and FOXM1 staining are assigned using a semi-quantitative five-category grading system as previously described (Sun et al., 2018 (link)).
Guo E., Xiao R., Wu Y., Lu F., Liu C., Yang B., Li X., Fu Y., Wang Z., Li Y., Huang Y., Li F., Wu X., You L., Qin T., Lu Y., Huang X., Ma D., Mills G.B., Sun C, & Chen G. (2021). WEE1 inhibition induces anti-tumor immunity by activating ERV and the dsRNA pathway. The Journal of Experimental Medicine, 219(1), e20210789.
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6

Evaluating T Cell-Mediated Cancer Cell Killing

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Primary human T cells were activated using CD3 antibody (Abcam, UK) and CD28 antibody (Abcam, UK). Colon cancer cells and activated T cells were cocultured in 6-well plates, and the wells were then washed with PBS three times to remove T cells after a period of coculture. The surviving cancer cells were fixed and stained using crystal violet solution.
Liu Y., Xu L., Hao C., Wu J., Jia X., Ding X., Lin C., Zhu H, & Zhang Y. (2022). Identification and Validation of Novel Immune-Related Alternative Splicing Signatures as a Prognostic Model for Colon Cancer. Frontiers in Oncology, 12, 866289.
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7

Immunofluorescent Detection of CD3 in Orbital Tissues

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In order to determine CD3 expression in orbital tissues, immunofluorescence staining was performed. The OFs were fixed in 10% neutral buffered formalin and permeabilized by 0.3% Triton X-100 in PBS. Fixed tissue or cells were rinsed in PBS, incubated in blocking solution (1% BSA, 0.3% Triton X-100 in PBS) for 30 min, then incubated in CD3 antibody (1:10; Abcam) solution at 4 °C overnight. The tissue or cells were washed three times in PBS and incubated in Alexa-488-conjugated secondary antibody (Invitrogen) for 1 h at room temperature. Finally, the tissue or cells were rinsed twice in PBS, and counterstained with DAPI for 5 min, and images were taken by fluorescence microscope (Olympus).
Li H., Min J., Yang Y., Suo W., Wang W., Tian J, & Qin Y. (2023). TMEM2 inhibits the development of Graves’ orbitopathy through the JAK-STAT signaling pathway. The Journal of Biological Chemistry, 300(2), 105607.
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8

Activated PBMC-Induced Apoptosis in Cancer Cells

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Human peripheral blood mononuclear cells (PBMCs) were purchased from ORIBIOTECH (Shanghai, China). 12-well plates were used to culture SMMC-7721 or Huh-7 cells with a density of 1 × 105 cells per well. PBMCs were activated by co-culturing with 2 μg/mL CD3 antibody (Abcam, Cambridge, UK) and 1 μg/mL CD28 antibody (Abcam) for 24 h. Then the activated PBMCs was incubated with SMMC-7721 or Huh-7 cells with the proportion of 5:1 of for 72 h. Annexin V and propidium iodide were stained. Finally, SMMC-7721 or Huh-7 cells were used to analyse the apoptosis rate by flow cytometry.
He L., Xu K., Niu L, & Lin L. (2022). Astragalus polysaccharide (APS) attenuated PD-L1-mediated immunosuppression via the miR-133a-3p/MSN axis in HCC. Pharmaceutical Biology, 60(1), 1710-1720.
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9

Investigating Inflammatory Responses

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QianLieJinDan tablets (National Drug Approval Number: B20020221) were obtained from Shandong Zhongda Pharmaceutical. Complete Freund's adjuvant (CFA) (Biyuntian Biotechnology, P2036), DAPI (Biyuntian Biotechnology, C1002), anti-fluorescence quenching sealing agent (Biyuntian Biotechnology, P0126), CD3+ antibody (Abcam, ab16669), DHE staining solution (Sigma, D7008), IL-6 kit (ELISA Biotech, EIA-3756), and Phospho-Stat3 (Ser727) Antibody(Cell Signaling Technology, #9134) were used.
Jia Z., Lv D., Chen T., Shi Z., Li X., Ma J., Gao Z, & Zhong C. (2024). Network pharmacology and in vivo experiment-based strategy for investigating the mechanism of chronic prostatitis/chronic pelvic pain syndrome in QianLieJinDan tablets. Heliyon, 10(9), e29975.
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