rat igg anti cd3  (Bio-Rad)

Journal: Regenerative Therapy

Article Title: Xenotransplantation of cryopreserved human clumps of mesenchymal stem cells/extracellular matrix complexes pretreated with IFN-γ induces rat calvarial bone regeneration

doi: 10.1016/j.reth.2022.04.003

Figure Lengend Snippet: Human C-MSCsγ and Cryo-C-MSCsγ suppressed rat T cells infiltration in a rat calvarial defect model. (A and B) Cultured human C-MSCs, C-MSCsγ, Cryo-C-MSCs, Cryo-C-MSCsγ were transplanted into a rat calvarial defect 1.6 mm in diameter with no artificial scaffold. Animals were sacrificed at 3 weeks after surgery, and the calvarial bones were fixed. Coronal serial sections were obtained and stained with HE (A) or immunostained with anti-rat CD3 antibody (B). Nuclei were counterstained with DAPI. The upper panel indicates the lower magnification images. Higher magnified images in the boxed regions are shown in the lower panels. (A) Bar = 500 μm (upper panels) and 250 μm (lower panels). (B) Bar = 250 μm (upper panels) and 50 μm (lower panels). (C) Graph shows the number of rat CD3-positive cells in the total defect area. Values are mean ± S.D. of six mice per group. ∗∗ p < 0.01 (ANOVA).

Article Snippet: These sections were incubated with a mouse anti-rat CD3 IgG antibody (clone G4.18; 1:100; Novus Biologicals, Littleton, CO) or a rabbit anti-human vimentin IgG antibody (clone SP20, 1:100, Abcam, Cambridge, MA) at 4 °C overnight.

Techniques: Cell Culture, Staining

Journal: PLoS ONE

Article Title: Pharmacologic Stem Cell Based Intervention as a New Approach to Osteoporosis Treatment in Rodents

doi: 10.1371/journal.pone.0002615

Figure Lengend Snippet: (A) Mouse BMMSCs co-cultured with LN cells indicated that anti-CD3 antibody activated LN cells were capable of inducing BMMSC death as shown a blank well without BMMSC staining (blue). When co-cultured BMMSCs (MSC) and LN cells were separated by a transwell culture system, anti-CD3 antibody treated LN cells failed to induce BMMSC death. (B) It is known that immunocompromised mice have no T lymphocytes. Thus, LN cells derived from immunocompromised failed to induce BMMSC death following anti-CD3 antibody activation in the co-culture system. (C) TUNEL staining showed that BMMSC death caused by anti-CD3 antibody-activated LN cell is through an apoptotic pathway. (D) Condition medium (CM) derived from naïve LN cells and anti-CD3 antibody activated LN cells were not able to induce cell death of BMMSCs. (E) Neutralizing anti-TNF-α and IFN-γ antibodies were not able to inhibit BMMSC death induced by anti-CD3 antibody-activated LN cells. (F) Neutralizing Fas ligand antibodies and brefeldin A, but not concanamycin A, were capable of blocking BMMSC death induced by anti-CD3 antibody-activated LN cells. (G) Western blot analysis showed that mouse and human BMMSCs (mMSC and hMSC) express Fas. (H) Fas antibody can induce significant reduction in number of living BMMSCs in culture. (I) Anti-CD3 antibody-activated LN cells were not able to induce cell death of BMMSCs derived from CD95 -deficient mice ( lpr ). (n = 5; [ P <0.05 and [[[ P <0.005).

Article Snippet: Rat anti-mouse CD3 IgG 3 , anti-mouse CD25 IgM, FITC- or PE-conjugated rat anti-mouse CD25 IgG 1 , FITC-, PE- or PerCP-conjugated rat anti-mouse CD4 IgG 2a , and PE-conjugated rat anti-mouse CD45R/B220 IgG 2a were purchased from BD Bioscience.

Techniques: Cell Culture, Staining, Derivative Assay, Activation Assay, Co-Culture Assay, TUNEL Assay, Blocking Assay, Western Blot

Journal: International Journal of Molecular Sciences

Article Title: Blockade of Glycosphingolipid Synthesis Inhibits Cell Cycle and Spheroid Growth of Colon Cancer Cells In Vitro and Experimental Colon Cancer Incidence In Vivo

doi: 10.3390/ijms221910539

Figure Lengend Snippet: The number of colorectal adenomas were reduced in Genz-treated mice. ( A ) Ki67 staining indicated proliferative cells in colons of mice. Both Genz-treated mice and controls developed predominantly adenomas with its typical polypoid-like shape ( A , B , middle images) and only low numbers of adenocarcinomas ( A , B, lower images). However, Genz-feeding markedly reduced the number of adenomas as compared to chow-fed controls ( B ). ( C ) The number of Ki67 positive cells in the colorectal parts of the colons was lower in crypts of Genz-treated mice as compared to controls; each data point represents mean numbers of Ki67 positive cells of approximately 50 crypts per mouse. Counted were positive cells only from completely vertically cut villi apart from the tumor areas; significances are *, p ≤ 0.05; ***, p < 0.001. ( D , F4/80) Both Genz-fed mice and controls lacked macrophage infiltrations in the tumor but showed an enrichment in peritumoral areas. ( D , CD3) Only a few cells stained positive with the T cell marker CD3; T, tumor area. Genz treatment did not lead to increased apoptosis ( D , TUNEL); a few cells with smaller nuclei than epithelial cells stained positive in tumors of treated mice and controls; scale bars, 100 µM.

Article Snippet: T cells were stained with rat anti-CD3 (#DIA-303, Hamburg, Germany), 1:50 and secondary biotinylated anti-rat (Vector), 1:200 and streptavidin- AP (Vector, Burlingame, CA, USA), 1:200.

Techniques: Staining, Marker, TUNEL Assay