Anti cd33

Manufactured by Thermo Fisher Scientific

Anti-CD33 is a laboratory reagent used for the detection and analysis of CD33 antigen expression on cells. It is commonly used in flow cytometry applications.

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Anti-human CD33 eFluor 450 (3 citations) Anti‐CD33 PE‐Cy7 (2 citations) Anti-CD33 clone WM53 (2 citations)

3 protocols using anti cd33

Molecular Mechanisms in Inflammatory Signaling

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The antibodies included anti-S100A9 (Cat no. ab92507; Abcam), anti-TLR4 (Cat no. sc-293072; Santa Cruz Biotechnology), anti-RAGE (Cat no. sc-80653; Santa Cruz Biotechnology), anti-p38 (Cat no. 9212; Cell Signaling Technology), anti-p65 (Cat no. 3034; Cell Signaling Technology), anti-ERK1/2 (Cat no. 4695; Cell Signaling Technology), anti-JNK (Cat no. 9253; Cell Signaling Technology), anti-AKT (Cat no. 8596; Cell Signaling Technology), anti-phospho(p)-p38 (Cat no. 4511; Cell Signaling Technology), anti-p-p65 (Cat no. 3033; Cell Signaling Technology), anti-p-ERK1/2 (Cat no. 3510; Cell Signaling Technology), anti-p-JNK (Cat no. 4668; Cell Signaling Technology), anti-p-AKT (Cat no. 9271; Cell Signaling Technology), anti-CD8 (Cat no. 340046, BD), anti-HLA-DR (Cat no. 4310370, eBioscience), anti-CD33 (Cat no. 4296343, eBioscience) and CD11b (Cat no. 4291932, eBioscience), and horseradish peroxidase-conjugated anti-mouse, anti-rabbit IgG antibodies. The inhibitors contained TAK-242 (MedChemExpress, New Jersey), FPS-ZM1 (MedChemExpress, New Jersey), SB203580 (Beyotime) and BAY 11-7082 (Beyotime). The preparation of the recombination GST-S100A9 protein, as well as its control protein GST, have previously been described (21 (link)).

Huang M., Wu R., Chen L., Peng Q., Li S., Zhang Y., Zhou L, & Duan L. (2019). S100A9 Regulates MDSCs-Mediated Immune Suppression via the RAGE and TLR4 Signaling Pathways in Colorectal Carcinoma. Frontiers in Immunology, 10, 2243.

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Myeloid cell differentiation and activation

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PBMC were isolated from healthy human donor blood buffy coats (New York Blood Center) by Lymphoprep gradient centrifugation (StemCell Technologies), according to manufacturer’s instructions. Myeloid cells were positively selected from PBMC using anti-human CD33 microbeads (Miltenyi Biotec) then cultured in complete RPMI-1640 medium for 5 days in the presence of GA and 50 ng/mL M-CSF (Peprotech) or 50 ng/mL GM-CSF (Peprotech) where indicated. Five days later, viable cells were analyzed by flow cytometry using anti-CD33, anti-CD14, anti-CD16, anti-CD163, anti-CD206, anti-CD86, and isotype-control matched antibodies (eBioscience). An aliquot of the 5-day culture was restimulated with LPS (Sigma-Aldrich) at 50 ng/mL for 16 hours followed by ELISA analysis of the supernatant.

van der Touw W., Kang K., Luan Y., Ma G., Mai S., Qin L., Bian G., Zhang R., Mungamuri S.K., Hu H.M., Zhang C.C., Aaronson S.A., Feldmann M., Yang W.C., Chen S.H, & Pan P.Y. (2018). Glatiramer acetate enhances myeloid-derived suppressor cell function via recognition of paired immunoglobulin-like receptor-B. Journal of immunology (Baltimore, Md. : 1950), 201(6), 1727-1734.

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Immunophenotyping of Mononuclear Cells

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Mononuclear cells were isolated from peripheral blood with Ficoll-Plaque PLUS (GE Heathcare) and blocked in human IgGs as described above. The mouse monoclonal anti-GRP78 (clone [10c3], Abcam) and control isotype antibodies were conjugated to PE with the Lightning-Link R-Phycoerythrin kit (Innova Biosciences). Cells were double-stained with anti-CD33 (conjugated to PercP-Cy5.5) and anti-CD34 (conjugated to allophycocyanin, APC) antibodies (both eBioscience) for 1 h in the dark and on ice. Cells were analyzed with a BD Accuri C6 flow cytometer. Data were processed with FlowJo software package (V.10.0.8, FlowJo LLC).

Staquicini D.I., D’Angelo S., Ferrara F., Karjalainen K., Sharma G., Smith T.L., Tarleton C.A., Jaalouk D.E., Kuniyasu A., Baze W.B., Chaffee B.K., Hanley P.W., Barnhart K.F., Koivunen E., Marchiò S., Sidman R.L., Cortes J.E., Kantarjian H.M., Arap W, & Pasqualini R. (2017). Therapeutic targeting of membrane-associated GRP78 in leukemia and lymphoma: preclinical efficacy in vitro and formal toxicity study of BMTP-78 in rodents and primates. The pharmacogenomics journal, 18(3), 436-443.

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