The anti-CTLA-4 Nb (Nb36) wasdeveloped in our previous study [17 (link)]. Anti-CTLA-4 mAb (Rabbit monoclonal, UC10-4F10–11), anti-CD3 mAb (Rabbit monoclonal, SP7), anti-CD56 mAb (Rabbit monoclonal, EPR21827), anti-CD80 mAb (Rabbit monoclonal, EPR1157(2)), anti-CD83 mAb (Rabbit monoclonal, EPR23809–19) and anti-MHC II mAb (Rabbit monoclonal, 6C6) were all purchased from Abcam (Cambridge, UK).
Anti cd3 mab
Manufactured by Abcam
Sourced in United Kingdom
Anti-CD3 mAb is a monoclonal antibody that binds to the CD3 complex, which is expressed on the surface of T cells. This antibody is commonly used in immunological research applications as a tool to activate and study T cell function.
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Lab products found in correlation
Anti ctla 4 mab, by Abcam (2 mentions)
Anti cd56 mab, by Abcam (2 mentions)
Anti cd80 mab, by Abcam (2 mentions)
Anti cd83 mab, by Abcam (2 mentions)
Anti mhc 2 mab, by Abcam (2 mentions)
Lymphocyte separation solution, by Absin (1 mentions)
Click it plus tunel assay, by Thermo Fisher Scientific (1 mentions)
Anti ki 67 antibody, by Maixin Group (1 mentions)
Rabbit anti rat igg, by Agilent Technologies (1 mentions)
Paraffin, by Sakura Finetek (1 mentions)
5 protocols using anti cd3 mab
1
Antibody Toolkit for Cellular Analysis
2
Generation and Characterization of Cytokine-Induced Killer Cells
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The spleens of BALB/c mice were obtained under aseptic conditions for the preparation of single-cell suspensions. The lymphocytes were separated using a lymphocyte separation solution (Absin, Shanghai, China) and washed twice in serum-free RPMI-1640 medium. The cell concentration was adjusted to 1×106/mL, cells were placed in a culture flask. medium containing 10% fetal bovine serum was added, and the cells were then cultured in a humidified atmosphere at 37°C with 5% CO2. Recombinant mouse interferon-γ (1000 U/mL, Boster, Wuhan, China) was added on the same day. After 24 h, 50 ng/mL anti-CD3 mAb (Abcam, Cambridge, UK), 100 U/mL recombinant mouse interleukin-1β (rm IL-1β) (Boster, Wuhan, China), and 300 U/mL recombinant mouse interleukin-2 (rm IL-2) (Boster, Wuhan, China) were added. The medium was changed every 3 days, and cytokines were added to maintain the cell concentration at 1-2×106 cells/mL. An inverted microscope was used to assess CIK growth, proliferation, and contamination. Fungal, bacterial culture, and endotoxin tests were performed before collection of CIK cells to confirm that these exogenous factors were not present. The survival rate of CIK cells was >97%, and their recovery survival after freezing was >75%.
3
Intratumoral Cell Proliferation and Apoptosis
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Mice from the above therapeutic experiment were sacrificed. The tumor tissues were separated, fixed in 4% paraformaldehyde, processed, and embedded in paraffin. To evaluate the intratumoral cell proliferation and apoptosis, the ultrathin tissue sections were either incubated with an anti-Ki-67 antibody (1:200 dilution; Maixin Biotechnology, China)-based IHC for cell proliferation, or subjected to a TUNEL assay (Click-iT Plus TUNEL assay, Invitrogen, Carlsbad, CA) for cell apoptosis, by following the manufacturer’s instructions. Similarly, the density of tumoral microvessels (CD31+) and the infiltration of T cells (CD3+) were quantified by IHC based on an anti-CD31 mAb (1:500; Abcam, Britain) and an anti-CD3 mAb (1:500; Abcam, Britain) as primary antibodies, following an HRP-labeled IgG monoclonal antibody as a secondary antibody staining, and the sections were counterstained with DAB. The number of positive cells was counted and analyzed to evaluate the density of intratumoral proliferated cells, apoptotic cells, microvessels or the T cell infiltration among different treatments.
4
Histological Analysis of Islet Transplants
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STZ-induced diabetic B6 mice were implanted in the left renal subcapsular space with 300 MMC-treated or nontreated islets cultured for 3 d. Mice with normalized blood glucose levels were killed on days 4 or 8 after grafting. Specimens of the transplanted kidney containing the islet graft were fixed in 10% formalin (GraphPad, La Jolla, CA, USA), embedded in paraffin (Sakura Finetek Japan Co, Tokyo, Japan), and serially cut into 4-µm thick sections. The sections were stained with hematoxylin and eosin (H&E) to estimate the host’s cell-mediated immune response. Subsequently, monocytes, T cells, and B cells that migrated to the transplantation site were subjected to immunohistochemical analyses.
To detect monocytes and T cells, we used anti-F4/80 mAb (1:200 dilution) and anti-CD3 mAb (1:100 dilution), each purchased from Abcam, and a rabbit anti-rat IgG (DAKO, Glostrup, Denmark) secondary antibody. For the detection of B cells, we used biotin-conjugated rat anti-mouse CD45 R mAb (1:50 dilution; Thermo Fisher Scientific Inc, Waltham, MA, USA) and rabbit anti-rat IgG (DAKO).
To detect monocytes and T cells, we used anti-F4/80 mAb (1:200 dilution) and anti-CD3 mAb (1:100 dilution), each purchased from Abcam, and a rabbit anti-rat IgG (DAKO, Glostrup, Denmark) secondary antibody. For the detection of B cells, we used biotin-conjugated rat anti-mouse CD45 R mAb (1:50 dilution; Thermo Fisher Scientific Inc, Waltham, MA, USA) and rabbit anti-rat IgG (DAKO).
5
Antibody Panel for Immune Profiling
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The anti-CTLA-4 Nb (Nb36) and anti-CD105 Nb were developed in our laboratory.
Anti-CTLA-4 mAb, anti-CD3 mAb, anti-CD56 mAb, anti-CD80 mAb, anti-CD83 mAb and anti-MHC II mAb were purchased from Abcam (Cambridge, UK).
Anti-CTLA-4 mAb, anti-CD3 mAb, anti-CD56 mAb, anti-CD80 mAb, anti-CD83 mAb and anti-MHC II mAb were purchased from Abcam (Cambridge, UK).
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