Cd33 pe

Manufactured by BD

Sourced in United States

CD33-PE is a fluorescent-labeled monoclonal antibody used for the identification and enumeration of CD33-positive cells in biological samples by flow cytometry.

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Lab products found in correlation

Cd14 pe, by BD (6 mentions) Cd34 apc, by BD (3 mentions) Cd45 apc cy7, by BD (3 mentions) Mcd45 apc cy7, by BD (3 mentions) Cd45 apc h7, by BD (3 mentions) Cd11b apc, by Thermo Fisher Scientific (3 mentions) Apc cd13, by Thermo Fisher Scientific (3 mentions) Percp cyanine 5.5 human cd45, by Thermo Fisher Scientific (3 mentions) Cd45 fitc, by BD (2 mentions) Cd3 cd19 cd56 apc, by BD (2 mentions) Cd15 fitc, by BD (2 mentions) Cd80 fitc, by BD (2 mentions) Hla dr fitc, by BD (2 mentions) Cd123 pe, by BD (2 mentions) Cd14 apc, by BD (2 mentions) Cd45 apc, by BD (2 mentions) Cd38 apc, by BD (2 mentions) Cytoflex lx flow cytometer, by Beckman Coulter (2 mentions) Sytox red, by Thermo Fisher Scientific (2 mentions) Cd41 pe cy7, by BioLegend (2 mentions)

Spelling variants of this product

CD33 PE-Cy7 (11 citations) CD33 PE (WM53, (5 citations) PE Mouse Anti-Human CD33 (4 citations) PE-conjugated anti-CD33 (4 citations) Anti-CD33-PE-Cy7 (4 citations) Anti-CD33 PE (3 citations) CD33-PE-Cy7 (clone P67.6; (2 citations) Anti Human CD33 PE (2 citations) CD33-PE-CF594 (clone WM53, (2 citations) Anti-CD33 PE (WM53, (2 citations)

32 protocols using cd33 pe

Multiparametric Flow Cytometry Analysis

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Cells were washed in phosphate-buffered saline and incubated for 30 minutes at 4 °C with a combination of monoclonal antibodies against human cell surface antigens as presented in Table 1. Antibodies were purchased as described: CD36 APC, CD14 PE-Cy7, CD42 PE, CD33 PE, CD34 APC, CD45 FITC (all from Becton Dickinson), CD34 Pacific Blue, CD45 Alexa Fluor 700, CD41 APC, CD90 APC, CD10 PE-Cy7, CD19 PE-Cy7 (all from Biolegend, Saint Quentin en Yvelines, France), and CD235a PE, 7-AAD (both from Beckmann Coulter, Villepinte, France). Cells were analyzed on a FACSCanto II flow cytometer (Becton Dickinson) using Diva software (version 6.1.3; Becton Dickinson).

Faivre L., Parietti V., Siñeriz F., Chantepie S., Gilbert-Sirieix M., Albanese P., Larghero J, & Vanneaux V. (2016). In vitro and in vivo evaluation of cord blood hematopoietic stem and progenitor cells amplified with glycosaminoglycan mimetic. Stem Cell Research & Therapy, 7, 3.

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Phenotypic Characterization of Monocyte-Derived Dendritic Cells

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Qualitative determinations of specific sub-populations were performed using fluorescent-labeled antibodies and flow cytometry. The purity of the elutriated monocytes was evaluated by flow cytometry using CD33-PE, CD15-FITC, CD3/CD19/CD56-APC and CD45-APC-Cy7 (Becton Dickinson, Mountain View, CA) and isotype controls (Becton Dickinson). The analysis of mDCs was undertaken after harvest on day 4. This included the standard “DC panel” adopted in our institution as lot release for mDCs products and other investigational markers. The panel consisted of CD86-FITC, CD83-PE, CD14-APC, CD209-FITC, CCR7-PE, CD40-APC, HLA-DRFITC, CD123-PE, CD11c-APC, CD80-FITC, CD154-PE, CD54-APC, CD16-FITC, CCR7-PE and CD1a-APC. The expression of HER2/neu was assessed by flow cytometry with anti-CD340 antibodies (BioLegend, San Diego, CA). Flow cytometry acquisition and analysis were performed with FACScanto flow cytometer (Becton Dickinson and Company, Franklin Lakes, NJ) according to CPS procedures. Spectral overlap was electronically compensated using single color controls. Quality controls were run before each session according to internal quality control policy.

Jin P., Chen W., Ren J., Chen S., Wood L., Zhao Y., Remaley A., Pham C., Lian S., Liu S., Liu H., Highfill S., Berzofsky J.A, & Stroncek D. (2018). Plasma from Some Cancer Patients Inhibits Adenoviral Ad5f35 Vector Transduction of Dendritic Cells. Cytotherapy, 20(5), 728-739.

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Comprehensive Immunophenotyping of Elutriated Monocytes and Dendritic Cells

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Analysis of expression of surface markers was performed using
fluorescent labeled antibodies (Abs) and flow cytometry. The purity of the
elutriated monocytes was assessed by flow cytometry using CD33-PE, CD15-FITC,
CD3/CD19/CD56-APC and CD45-APC-Cy7 (Becton Dickinson, Mountain View, CA, USA)
and isotype controls (Becton Dickinson). DC were analyzed after pulsing on Day
4. The analysis included the standard “DC panel” adopted in our
institution as lot release for mature DC products and other investigational
markers. The panel consisted of CD86-FITC, CD83-PE, CD14-APC, HLA-DR-FITC,
CD123-PE, CD11c-APC, CD80-FITC, CD54-APC, CCR7-APC, and CD38-FITC (Becton
Dickinson). Flow cytometry acquisition and analysis were performed with a
FACScanto flow cytometer (Becton, Dickinson and Company, Franklin Lakes, NJ USA)
according to CPS standard operating procedures. Spectral overlaps were
electronically compensated using single color controls. Quality controls were
run before each session according to internal quality control policy.

Castiello L., Sabatino M., Ren J., Terabe M., Khuu H., Wood L.V., Berzofsky J.A, & Stroncek D.F. (2017). Expression of CD14, IL-10 and tolerogenic signature in dendritic cells inversely correlate with clinical and immunological response to TARP vaccination in prostate cancer patients. Clinical cancer research : an official journal of the American Association for Cancer Research, 23(13), 3352-3364.

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Immunophenotypic Analysis of Leukemic Engraftment

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CD45-APC (BD, San Jose, CA, USA; 555485), CD33-PE (BD 555450), CD44-FITC (BD 555478), CD44-PE-Cy7 (BD 560533), CD45.1-PerCP-Cy5.5 (BD 560580), CD34-APC-Cy7 (Biolegend, Ozyme, Saint Quentin Yvelines, France; 343514), Lin1-FITC (BD 340546), CD123-PE (BD 340545), CD45RA-Alexa Fluor 700 (BD 560673) and CD38-APC (BD 555362) fluorescent antibodies were used to analyze leukemic cells before and after injection into animals to determine phenotypic analysis of engrafted cells and percentage of leukemic cell engraftment. Absolute cell counts were determined with CountBright absolute counting beads (Invitrogen) following the manufacturer's recommendations.

Saland E., Boutzen H., Castellano R., Pouyet L., Griessinger E., Larrue C., de Toni F., Scotland S., David M., Danet-Desnoyers G., Vergez F., Barreira Y., Collette Y., Récher C, & Sarry J.E. (2015). A robust and rapid xenograft model to assess efficacy of chemotherapeutic agents for human acute myeloid leukemia. Blood Cancer Journal, 5(3), e297-.

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Engraftment of CRISPR-modified HSPCs

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All mouse experiments were approved by the Austrian Ministry of Education, Science and Research (BMBWF-66.010/0056-V/3b/2019). 5x10⁵ CRISPR-modified HSPCs were intrafemorally transplanted into irradiated 8–12-week-old NSG mice. For competitive transplants, 1x10⁵ sort-purified HSPCs (1:1 mixture of CALR INS and WT cells) were intrahepatically transplanted into new-born NSGW41 mice. Human engraftment in murine bone marrow was evaluated after 8, 16 and 24 weeks via flow cytometry. Cells were stained with mTer119-BUV661, mCD45-APC-Cy7, hCD45-BB700, CD33-PE (BD Biosciences), CD19-SB600 (eBioscience, San Diego, CA, USA), CD41-PE-Cy7 (BioLegend) antibodies and SYTOX Red (Invitrogen, Carlsbad, CA, USA) and measured on a CytoFLEX LX flow cytometer (Beckman Coulter).

Foßelteder J., Pabst G., Sconocchia T., Schlacher A., Auinger L., Kashofer K., Beham-Schmid C., Trajanoski S., Waskow C., Schöll W., Sill H., Zebisch A., Wölfler A., Thomas D, & Reinisch A. (2023). Human gene-engineered calreticulin mutant stem cells recapitulate MPN hallmarks and identify targetable vulnerabilities. Leukemia, 37(4), 843-853.

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Multifaceted Cellular Analysis by FACS

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Analysis of samples was performed on a LSR2 and Fluorescence-activated Cell Sorting(FACS) on an ARIA or Fusion-instrument (BD Biosciences, San Jose, CA, USA). Single cells were gated on the basis of FSC-Height(FSC-H) and FSC-Area(FSC-A), and live cells were gated out using Propidium Iodide(PI) or the LIVE/DEAD™ Fixable Aqua Dead Cell Stain Kit (ThermoFisher Scientific, Waltham, MA, USA). Antibodies were obtained from BioLegend (San Diego, CA, USA): CD1a PB, CD3 Pe-Cy7, CD3 APC-Cy7, CD3 BV421, CD4 PerCp-Cy5.5, CD5 PE, CD8 Amcyan, CD8 APC-Cy7, CD14 PB, CD19-APC, CD27 APC-Cy7, CD33 PE, CD34 PerCp-Cy5.5, CD34 APC, CD45 Amcyan, CD45 Pe-Cy7, CD56 BV421, CD69 Pe-Cy7, CD137 PE, interferon-γ PE, IL-2 PB, TCRαβ-PE, and TCRγδ APC; BD Biosciences (San Jose, CA, USA): CD7-FITC; ThermoFisher Scientific (Waltham, MA, USA): CellTrace™ Violet Cell Proliferation Kit; and SCBT (Dallas, TX, USA): WASp-AF647.

Pille M., Avila J., Sanchez G.S., Goetgeluk G., De Munter S., Jansen H., Billiet L., Weening K., Xue H., Bonte S., Ingels J., De Cock L., Pascal E., Deseins L., Kerre T., Taghon T., Leclercq G., Vermijlen D., Davis B, & Vandekerckhove B. (2023). The Wiskott–Aldrich syndrome protein is required for positive selection during T-cell lineage differentiation. Frontiers in Immunology, 14, 1188099.

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Immune Cell Isolation and Characterization

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Citric acid (ACS reagent, ≥99.5%), Diethylentriamine (DETA, 99%), Dulbecco's Phosphate Buffered Saline (DPBS), Lysis Buffer, Float-A-Lyzer dialysis devices (100–500 Da). Antibodies against CD45-PE-Cy7, CD34-PerCP-Cy5.5, CD33-PE, CD19-APC-R700 and CD3-APC-H7 were purchased from BD biosciences. Stem SPAN™ SFEM medium was bought at STEMCELL™ Technologies and microwave reaction vessels were obtained from CEM GmbH. NucBlue™ Live ReadyProbes™ Reagent (Invitrogen™), Poly-l-Lysine coated 8 well μ-slide was obtained from Ibidi.

Nollmann C., Wimmenauer C., Fasbender S., Mayer S., Caddedu R.P., Jäger P., Heinzel T, & Haas R. (2021). Uptake of carbon nanodots into human AML cells in comparison to primary hematopoietic cells. RSC Advances, 11(42), 26303-26310.

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Multiparametric Flow Cytometry Analysis

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Multi-color flow cytometry analysis was performed on PBMCs from all time points by staining for 30 minutes at 4°C with CD3-V450, CD8-FITC or APC, ICOS-PE, HLA-DR-PerCP-Cy5.5, CD25-PE-Cy7, CD45RA-PerCP-Cy5.5, CD62L-FITC, CD127-V450, PD-1-PE, Tim-3-AF700, CD4-APC-Cy7 (BD Biosciences, San Jose, CA), CCR7-PE-Cy7 (R&D Systems, Minneapolis, MN), CTLA-4-FITC (LSBio, Seattle, WA) and FoxP3-APC (eBioscience, San Diego, CA) for T cells. For natural killer (NK) cells, CD3-V450, CD16-APC-Cy7, CD56-PE-Cy7 and Tim-3-AF700 (BD) were used. For myeloid-derived suppressor cells (MDSCs), CD33-PE, CD11b-APC-Cy7, HLA-DR-PerCP-Cy5.5, CD14-V450 and CD15-APC (BD) were used. 1×10⁵ cells were acquired on an LSRII (BD), and data was analyzed using FlowJo software (Tree Star Inc., Ashland, OR). The appropriate isotype controls were used, and dead cells were excluded from the analysis.

Jochems C., Tucker J.A., Tsang K.Y., Madan R.A., Dahut W.L., Liewehr D.J., Steinberg S.M., Gulley J.L, & Schlom J. (2014). A Combination Trial of Vaccine Plus Ipilimumab in Metastatic Castration-Resistant Prostate Cancer Patients: Immune Correlates. Cancer immunology, immunotherapy : CII, 63(4), 407-418.

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Multiparametric Flow Cytometry Analysis of AML and HSPCs

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For analysis and sorting of AML and HSPCs derived from hCB or adult BM, cells were stained with hCD45-PeCy7(Clone: H30, cat: 560915, dilution 1 in 25), mCD45-PerCPCy5.5 (Clone:30-F11, cat: 550994, dilution 1 in 400), Lineage-FITC (lin1, cat: 340546, 1 in 25), CD34-PE (Clone 581, cat: 560941, dilution: 1 in 25), and CD38-APC (Clone HIT2, cat: 555462; dilution: 1 in 25) (BD Biosciences, Oxford, UK). Human grafts in mice were assessed using CD19-FITC (Clone: H1B19; cat: 555412, dilution: 1 in 25), CD33-PE (Cat: WM53, cat: 555450, dilution 1 in 25), CD3-APC (Clone: UCHT1, cat: 561811, dilution: 1 in 25), hCD45-PeCy7 (Clone: H30, cat: 560915, dilution 1 in 25), and mCD45-PerCPCy5.5 (Clone:30-F11, cat: 550994, dilution 1 in 400) (BD Biosciences). Luciferase-transduced HL60 and U937 cells were identified and sorted based on their GFP expression. Non-viable cells were excluded by DAPI staining. Appropriate isotype-matched antibodies were used as controls. Flow cytometry analysis was performed using an LSRII flow cytometer (BD Biosciences). Cell sorting was performed using a FACS Aria or INFLUX (BD Biosciences).

Di Tullio A., Rouault-Pierre K., Abarrategi A., Mian S., Grey W., Gribben J., Stewart A., Blackwood E, & Bonnet D. (2017). The combination of CHK1 inhibitor with G-CSF overrides cytarabine resistance in human acute myeloid leukemia. Nature Communications, 8, 1679.

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Engraftment and Leukemia Analysis

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Tail bleeds were analyzed on a Hemavet9500 (Drew Scientific). Engraftment was determined from flow cytometry of PB and BM preparations using a FACSCantoII instrument (BD) with analysis by FlowJo software. Our standard flow panel consists of antibodies to block mouse and human Fc IgG receptors (Miltenyi Biotech) as well as mCD45-APC/Cy7(BD), CD45-FITC (BD), CD3-PE/Cy7 (BD), CD19-VioBlue (Miltenyi Biotech), CD13-PE (BD), CD33-PE (BD), CD34-APC (BD), and CD56-v510 (BD). Leukemia percentage was determined by calculating the number of cells with positive staining for CD33 and CD45 (AML) or CD19 and CD45 (ALL) as a fraction of viable cells.

Wunderlich M., Manning N., Sexton C., Sabulski A., Byerly L., O’Brien E., Perentesis J.P., Mizukawa B, & Mulloy J.C. (2019). Improved chemotherapy modeling with RAG-based immune deficient mice. PLoS ONE, 14(11), e0225532.

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