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Stainrite wright giemsa stain solution

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StainRITE Wright-Giemsa Stain Solution is a pre-formulated staining solution. It is used for the differential staining of blood cells and other cellular components in medical and research laboratory applications.

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Lab products found in correlation

Poly mount, by Polysciences (2 mentions) Lsrfortessa, by BD (2 mentions) Nunc lab tek 2 chamber slide, by Thermo Fisher Scientific (1 mentions) Pe labeled anti cd11b, by BioLegend (1 mentions) Facsaria 3, by BD (1 mentions) K562 cell line, by American Type Culture Collection (1 mentions)

4 protocols using stainrite wright giemsa stain solution

1

Monocytic Differentiation Induced by PMA

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Cells were cultured at 1×105 cells/well (1 ml) in 6-well plates for flow cytometry, or in slide chambers (Nunc Lab-Tek II Chamber Slide; Thermo Fisher Scientific, Inc.) for Wright-Giemsa staining, and were subjected to treatment with various concentrations of PMA (0, 1, 5, 10, 50, 100 and 200 ng/ml). After 96 h, adhesive cells in 6-well plates were harvested using cell scrapers, washed with cold PBS and stained with anti-cluster of differentiation (CD)11b-phycoerythrin antibody (Ab) (clone M1/70, 101208; BioLegend, Inc.) at 0.02 mg/ml on ice for 30 min, followed by flow cytometry. For Wright-Giemsa staining, adherent cells in slide chambers were washed with PBS and stained with StainRITE Wright-Giemsa Stain Solution (Polysciences, Inc., Warrington, PA, USA) to examine their morphologies using a Zeiss Axioskope 2 Plus microscope (Carl Zeiss AG, Oberkochen, Germany) and to assess differentiation.
Wu Z.J., Zhao X., Banaszak L.G., Gutierrez-Rodrigues F., Keyvanfar K., Gao S.G., Raffo D.Q., Kajigaya S, & Young N.S. (2018). CRISPR/Cas9-mediated ASXL1 mutations in U937 cells disrupt myeloid differentiation. International Journal of Oncology, 52(4), 1209-1223.
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2

Cytospin and Flow Cytometry Analysis

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After counting with trypan blue exclusion, 20,000–50,000 cells were loaded and cytospins were prepared at 1000 rpm for 5 min. Slides were air-dried and stained with StainRITE Wright-Giemsa Stain Solution (Polysciences) and mounted with Poly-mount (Polysciences). For flow cytometric analysis, K562, NB4 or MOLM13 cells with different treatments were collected and washed with chilled PBS, followed by staining with APC-CD61 (eBioscience, cat. no. 17-0619-42), PE-labelled anti-CD11b (BioLegend, cat. no. 101208) and APC-labelled anti-CD14 (eBioscience, cat. no. 17-0149-41) antibodies for 25 min. Cells were then fixed and analysed on a BD LSRFortessa or FACSAria III analyser (BD Biosciences).
Dou X., Xiao Y., Shen C., Wang K., Wu T., Liu C., Li Y., Yu X., Liu J., Dai Q., Pajdzik K., Ye C., Ge R., Gao B., Yu J., Sun S., Chen M., Chen J, & He C. (2023). RBFOX2 recognizes N6-methyladenosine to suppress transcription and block myeloid leukaemia differentiation. Nature Cell Biology, 25(9), 1359-1368.
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3

Monocytic Cell Differentiation Assay

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After counting with trypan blue exclusion, 20000–50000 cells were loaded and cytospins were prepared at 1000 rpm for 5 min. Slides were let dry and stained with StainRITE® Wright-Giemsa Stain Solution (Polysciences, Warrington, PA) and mounted with Poly-mount (Polysciences).
For flow cytometric analysis, MM6, NB4 or U937 cells with different treatments were harvested and washed with chilled PBS, followed by staining with PE-labeled anti-CD11b (101208, BioLegend) and APC-labeled anti-CD14 (17-0149-41, eBioscience) antibodies for 25 minutes. Cells were then fixed and analyzed on a BD LSRFortessa or FACSAria III analyzer (BD Biosciences).
Weng H., Huang H., Wu H., Qin X., Zhao B.S., Dong L., Shi H., Skibbe J., Shen C., Hu C., Sheng Y., Wang Y., Wunderlich M., Zhang B., Dore L.C., Su R., Deng X., Ferchen K., Li C., Sun M., Lu Z., Jiang X., Marcucci G., Mulloy J., Yang J., Qian Z., Wei M., He C, & Chen J. (2017). METTL14 Inhibits Hematopoietic Stem/Progenitor Differentiation and Promotes Leukemogenesis via mRNA m6A Modification. Cell stem cell, 22(2), 191-205.e9.
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4

Karyotyping of K562 and DNMT3A KO Cells

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The K562 cell line and HAP1 GeneArt engineered DNMT3A KO cell line were purchased from the American Type Culture Collection (ATCC) and Thermo Fisher Scientific, respectively. All cell lines were cultured in IMDM medium supplemented with 10% fetal bovine serum and antibiotics and were incubated at 37°C in a humidified atmosphere of 95% air and 5% CO2. Cells in the logarithmic growth stage were cytospun on slides using the Shandon Cytospin 4 and subjected to staining with the StainRITE® Wright-Giemsa Stain Solution (Polysciences, Warrington, PA) to examine their morphologies. Standard G-band karyotype analysis was performed using passage-matched parental cells within 7 days of thawing (Karyologic, Inc., Durham, NC, USA).
Banaszak L.G., Giudice V., Zhao X., Wu Z., Gao S., Hosokawa K., Keyvanfar K., Townsley D.M., Gutierrez-Rodrigues F., del Pilar Fernandez Ibanez M., Kajigaya S, & Young N.S. (2018). Abnormal RNA splicing and genomic instability after induction of DNMT3A mutations by CRISPR/Cas9 gene editing. Blood cells, molecules & diseases, 69, 10-22.
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