mouse anti cd33 fitc  (Miltenyi Biotec)

Journal: Stem Cell Reports

Article Title: Cryopreserved Human Oocytes and Cord Blood Cells Can Produce Somatic Cell Nuclear Transfer-Derived Pluripotent Stem Cells with a Homozygous HLA Type

doi: 10.1016/j.stemcr.2020.05.005

Figure Lengend Snippet: The Establishment of a Humanized Mouse Model Was Confirmed by a Human Pan Hematopoietic Cell Markers (A) Although the frequencies of T lymphocytes were low, stable engraftment of human cells was revealed in tissues, including the PB, BM, and spleen. (B) Immunocytochemistry showed that CD45 was detected in the PB. Scale bars, 20 μm. (C) High engraftment of CD33 + myeloid lineage cells, compared with that of CD19 + and CD3 + lymphoid lineage cells, was revealed by FACS analysis. (D) In T cells, CD4 + CD8 + immature thymocyte precursors were regenerated in the mouse thymus and mature T cells, such as single CD4 + and CD8 + cells were detected in the PB, BM, and spleen at 18 weeks after transplantation of CD34 + cells.

Article Snippet: APC-conjugated mouse anti-human CD45 (BD), fluorescein isothiocyanate (FITC)-conjugated rat anti-mouse CD45 (BD), PerCP-conjugated mouse anti-human CD45RA (BioLegend), FITC-conjugated mouse anti-human CD45RO (BioLegend), PE-conjugated mouse anti-human CD19 (BioLegend), mouse anti-human TCRαβ (BioLegend), APC-conjugated mouse anti-human CD4 (BD), PE-conjugated mouse anti-human CD8 (BD), FITC-conjugated mouse anti-human CD33 (BD), rabbit anti-human MHC class I (Abcam), and mouse anti-human MHC class II (Abcam) were used as a primary antibodies.

Techniques: Immunocytochemistry, Transplantation Assay

Journal: Molecular and Cellular Biology

Article Title: SRSF1 and PTBP1 Are trans -Acting Factors That Suppress the Formation of a CD33 Splicing Isoform Linked to Alzheimer’s Disease Risk

doi: 10.1128/MCB.00568-18

Figure Lengend Snippet: The rs12459419 SNP genotype does not affect CD33 mRNA or protein levels in CD33 cDNAs lacking introns. (A) Overview of the CD33 expression constructs used. (B) Overview of the RT-qPCR primer and probe sets used to detect the various CD33 mRNA transcripts. (C) Validation of the RT-qPCR assays ability to quantify CD33 exon 2 splicing using gBlocks that represent D2-CD33 or full-length CD33 cDNA. The fraction of detected exon 2 included (red) or exon 2 skipped (cyan) gene fragments is shown. Input quantities of the two different CD33 fragments are indicated below the graph. (D and E) mRNA levels of exon 2 included (D) and exon 2 skipped (E) CD33 transcripts normalized to the corresponding T-allele in HeLa cells transfected with the CD33 constructs shown in panel A, measured by RT-qPCR. (F) The RT-PCR products from a single primer set that simultaneously detects full-length CD33 (FL) and D2-CD33 in HeLa cells transfected with the CD33 constructs, including introns (A), visualized on a 1.2% agarose gel. The percent D2-CD33 values ± the standard errors of the mean (SEM) are indicated below each lane. (G to L) Flow cytometry analysis of CD33 surface levels using FITC- or PE-labeled antibodies targeting exon 2 (WM-53 [G and H]) or exon 3 (Him3-4 [J and K]) of CD33 in HeLa cells transfected with the CD33 constructs shown in panel A. CD33 KO THP1 cells were utilized as a negative control. α IgV, anti-IgV; α IgC, anti-IgG C 2 domain. The mean fluorescence intensity (MFI) normalized to the corresponding T-allele is depicted in panels I and L. Error bars indicate means + the SEM. A two-tailed t test was performed. *, P ≤ 0.05; **, P ≤ 0.01; ****, P ≤ 0.0001; ns, not significant. n = 3 biological replicates for each plot.

Article Snippet: The antibodies used were as follows: phycoerythrin (PE)-conjugated mouse anti-human CD33 clone WM-53 (BD Biosciences), fluorescein isothiocyanate (FITC)-conjugated mouse anti-human CD33 clone him3-4 (BD Biosciences, catalog no. 555626), and Alexa Fluor 488-conjugated anti-human CD11b clone ICRF44 (BioLegend, catalog no. 301317).

Techniques: Expressing, Construct, Quantitative RT-PCR, Transfection, Reverse Transcription Polymerase Chain Reaction, Agarose Gel Electrophoresis, Flow Cytometry, Labeling, Negative Control, Fluorescence, Two Tailed Test