apc-cy7 cd45.1  (Becton Dickinson)

Journal: bioRxiv

Article Title: Epigenetic control of CD8 + T cell tissue homing and tissue resident memory T cell precursors by the histone methyltransferase SUV39H1

doi: 10.1101/2024.05.13.593994

Figure Lengend Snippet: A. Scheme of the experiment. B. Representative dot plots of WT (CD45.1-) and SUV39H1 KO (CD45.1+) OTI cells (Vα2+) in the indicated organs 33 days after adoptive transfer. C. Quantification of the ratio between SUV39H1 KO and WT OTI cells within each organ. D. Representative dot plot of intravascular (iv) staining ( left ) and expression of CD49d and Ly6c ( right ) in each population at day 34 post transfer in CD8 + T cells from lungs from Exp. 2 in C . E. Quantification of the percentage and absolute numbers of CD49d + Ly6c - cells among SUV39H1KO or WT OTI cells in the indicated organs from Exp 1 in C . Numbers represent the fold increase between KO and WT condition for each organ. F. Histogram of CD69 expression in iv-CD49d + Ly6c - CD8 + T cells and proportions of CD69+ cells among iv+ and iv-CD49d + Ly6c - CD8 + T cells in lungs from Exp. 4 in C . Numbers represent p values. Each dot represents a mouse, different shapes represent different experiments. * p<0.05 ; ** p<0.01 ; *** p< 0.001 ; **** p<0.0001 ; ns: not significant by one sample t test compared to 1 ( C ), multiple ratio paired t test with Holm-Šídák correction ( E ) and multiple paired t test ( F ).

Article Snippet: Cell suspensions were prepared in PBS and stained with Near IR (Fisher Scientific L34975) or Acqua LIVE/DEAD™ Fixable Cell Stain Kit (Fisher Scientific L34957) according to manufacturer’s instructions to exclude dead cells and then labelled in FACS buffer with the following antibodies: V500 Rat anti-Mouse CD4 (RM4-5, BD), eF450 CD5 Monoclonal Antibody (53-7.3, eBiosciences), FITC anti-CD8α (53-6.7, BD), PerCP-Cy5.5 CD8 α (53-6.7, BD), PE CD8 α Monoclonal Antibody (53-6.7, Ficher Scientific), BUV395 Rat Anti-Mouse CD8 β (H35-17.2, BD), APC anti-mouse CD25 Antibody (PC61, Biolegend), PerCP-Cy5.5 Rat Anti-Mouse CD44 (IM7, BD), APC Rat Anti-Mouse CD44 (IM7, BD), APC-R700 Rat Anti-Mouse CD44 (IM7, BD), eFluor 450 CD45.1 Monoclonal Antibody (A20, Fisher Scientific), PE Mouse Anti-Mouse CD45.1 (A20, BD), APC-Cy7 CD45.1 (A20, BD), PE Mouse Anti-Mouse CD45.2 (104, BD), APC Mouse anti-Mouse CD45.2 (104, BD), BV605 Hamster Anti-Rat/Mouse CD49a (Ha31/8, BD), BV786 Rat Anti-Mouse CD49d (R1-2, BD), PE/Cyanine7 anti-mouse CD49d Antibody (R1-2, Biolegend), PE-Cy7 Rat Anti-Mouse CD62L (MEL-14, BD), APC Rat Anti-Mouse CD62L (MEL-14, BD), APC/Cyanine7 anti-mouse CD62L Antibody (MEL-14, Biolegend), PerCP-Cy5.5 Hamster Anti-Mouse CD69 (H1.2F3, BD), PE-Cy7 Hamster Anti-Mouse CD69 (H1.2F3, BD), Brilliant Violet 605™ anti-mouse CD73 Antibody (TY/11.8, Biolegend), eFluor 450 CD103 Monoclonal Antibody (2E7, Fisher Scientific), eF660 CD127 (A7R34, eBiosciences), Brilliant Violet 711 anti-mouse CX3CR1 (SA011F11, Biolegend), BV711 CXCR3 (CXCR3-173, BD), Brilliant Violet 650™ anti-mouse Ly-6C Antibody (HK1.4, Biolegend), FITC anti-mouse Ly-6C (HK1.4, Biolegend), APC Ly-6C Monoclonal Antibody (HK1.4, eBiosciences), PE-CF594 Hamster Anti-Mouse KLRG1 (2F1, BD), BUV737 Hamster Anti-Mouse TCR β Chain (H57-597, BD), Alexa-Fluor®488 Hamster Anti-Mouse TCR β Chain (H57-597, BD), PE-Cy™7 Hamster Anti-Mouse TCR β Chain (H57-597, BD), PE-CF594 TCR β Chain (H57-597, BD), eFluor 450 TRCR V alpha 2 (B20.1, Invitrogen), PerCP/Cy5.5 anti-mouse TCR Vα2 (B20.1, Biolegend).

Techniques: Adoptive Transfer Assay, Staining, Expressing

Journal: Cell Stem Cell

Article Title: Controlling genetic heterogeneity in gene-edited hematopoietic stem cells by single-cell expansion

doi: 10.1016/j.stem.2023.06.002

Figure Lengend Snippet: Single-cell cloning of gene-edited functional HSCs (A) Schematic showing the extracellular domain of CD45 with allele-specific antibody clones 104 and A20 and the epitope-defining amino acid. (B) Experimental setup of the single-cell editing and expansion experiment. (C) Left: fractions of CD201 + CD150 + KSL cells in single-cell-derived cultures 14 days after cloning (n = 261 clones). Right: Histogram of CD201 + CD150 + KSL cell frequency. Zoomed-in region shows clones with >10% CD201 + CD150 + KSL cells. (D and E) CD45.1 + donor PB chimerism (D) and lineage distribution (E ) in single recipients with long-term (LT) engraftment ≥5% and multilineage reconstitution (n = 8). Numbers over graphs in (D) represent percentage of CD201 + CD150 + KSL cells in the transplanted clone (%). (F) Linear correlation plots of CD201 + CD150 + KSL cell frequency and 16-week donor chimerism. Red dots indicate LT repopulating and multilineage clones. Pearson correlation. (G) CD45.1 + PB chimerism and lineage distribution in secondary recipients (n = 5). See also Figure S4 and Table S3 . Error bars represent SD.

Article Snippet: anti-mouse CD45.1-APC/Cy7 (A20) , Tonbo Biosciences , Cat#25-0453; RRID: AB_2621629.

Techniques: Cloning, Functional Assay, Clone Assay, Derivative Assay

Journal: Cell Stem Cell

Article Title: Controlling genetic heterogeneity in gene-edited hematopoietic stem cells by single-cell expansion

doi: 10.1016/j.stem.2023.06.002

Figure Lengend Snippet:

Article Snippet: anti-mouse CD45.1-APC/Cy7 (A20) , Tonbo Biosciences , Cat#25-0453; RRID: AB_2621629.

Techniques: Recombinant, Sequencing, Software, CRISPR

Journal: STAR Protocols

Article Title: Assessing in vivo presentation of exogenous antigen in the tumor microenvironment in mice

doi: 10.1016/j.xpro.2023.102185

Figure Lengend Snippet:

Article Snippet: CD45.1 – APC-Cy7 (1:400) , Tonbo Biosciences , clone: A20; Cat# 25-0453-U100.

Techniques: Blocking Assay, Recombinant, Selection, Software, Cell Culture, Light Microscopy