Human Survivin Antibody

Detection of Survivin in Jurkat cells by Flow Cytometry

Jurkat cells were stained with Mouse Anti-Human Survivin Monoclonal Antibody (Catalog # MAB886, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram) followed by Phycoerythrin-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # F0102B). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.

Detection of Survivin by Flow Cytometry

Exposure to survivin enriched PD-1+ Bcl-6+ subset of Tfh via STAT3 dependent mechanismsDBA/1 mice were immunized with survivin-derived peptide (100 μg/mouse × 4, subcutaneously). Control mice were immunized with ovalbumin-derived peptide (OVA). Both groups were then subjected to collagen-induced arthritis. Anti-survivin IgG antibodies, anti-Fcg antibodies, & survivin levels in serum were measured by ELISA A. Flow cytometry analysis of the expression of survivin B. in CD44hiCD4+ lymphocytes, & expression of PD-1 C. & Bcl-6 E. in survivin+CXCR5+ CD4+ lymphocytes in spleen (SPL) & lymph nodes (LN). PD-1 expression correlated to the size of CXCR5+survivin+ population D. Dots represent individual mice & the horizontal line shows median of the group. Protein levels of active STAT3 phosphorylated at Y705 (pStat3), total Stat3, Bcl-6 & actin in spleen were analyzed by the Western blot F. The levels of each protein were quantified in ratio to actin of each sample. Quantification of the detected bands is presented as box plots. Transcription of Bcl-6, cMaf & IL-21 in the spleen was analyzed by RT-PCR & presented in relative quantity (RQ) to the median of the control group G. Anti-survivin antibodies in serum correlated with the size of survivin+ CD4 population in spleen of survivin-immunized mice H. Comparison of the correlations in the survivin- & OVA-immunized groups is done by the Fisher r-to-z transformation analysis. Anti-Fcg antibodies correlated with the intensity of PD-1 on CXCR5+survivin+ CD4 lymphocytes I., as measured by flow cytometry. Box plots with line represent IQR of the group & median, respectively, & error lines indicate min & max values. The Mann-Whitney U-test was used to compare differences between groups. Correlation analyses were performed using Spearman's test. Image collected & cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26343374), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Survivin by Flow Cytometry

Survivin positive subset of CD44hi CD4 lymphocytes in mouse possess a complete phenotype of Tfh cells Spleen and lymph nodes from collagen II immunized arthritic (CIA) mice were analyzed for expression of survivin and Bcl-6 using flow cytometry A. Cells were gated on memory CD44hiCD4+ lymphocytes. Expression of CXCR5 B. and PD-1 C. was investigated within Bcl-6+ survivin+ and Bcl-6− survivin+ cells. Dots represent individual mice and the horizontal line shows median of the group. Survivin translation in CIA mice was inhibited by shRNA-producing constructs provided as a single intra-peritoneal injection (shSurv16, n = 10, or shSurv13+16, n = 10). Control mice were treated with a non-targeting RNA construct (shNT, n = 9). Survivin expression in spleen was analyzed by flow cytometry 12 days after the injection. Cells were gated on CD4+ lymphocytes. Intensity of survivin expression (MFI) within the groups is shown by a representative histogram and summarized in a box plot D. Survivin expression (MFI) on CD4 lymphocytes correlated to the size of CD44hiCD62L+ population E. Expression of CXCR5 on CD44hiCD4+ lymphocytes in the groups is shown as box plot F. CXCR5+ population correlates with the intensity of survivin G. and with Bcl-6 mRNA H. Boxes and lines represent IQR and median, respectively, and error lines indicate min and max values. The Mann-Whitney U-test was used to compare differences between groups. Correlation analyses were performed using Spearman's test. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/26343374), licensed under a CC-BY license. Not internally tested by R&D Systems.