Ab27287

Manufactured by Abcam

Sourced in United States

Ab27287 is a laboratory product manufactured by Abcam. It is a core component used in various research and diagnostic applications.

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Lab products found in correlation

6 protocols using ab27287

Phenotypic Characterization of Human MSCs

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Human MSCs single-cell suspensions were harvested using a 0.05% trypsin/
Ethylenediaminetetraacetic acid (EDTA) solution; after FBS neutralization incubated in
blocking buffer [1% FBS in Dulbecco’s phosphatebuffered saline (DPBS)] for 30 minutes.
Next, 1×10⁶ cells were separately incubated for 1 hour at 4°C with an optimal
dilution of conjugated antibodies that included anti-CD73-FITC (ab28061), anti-CD45- FITC
(ab27287), anti-CD90-FITC (ab11155), antiCD34-PE (ab157304), and anti-CD105-PE (ab91138),
all from Abcam (Cambridge, UK). Flow cytometry experiments were performed with a BD
FACSCalibur Flow Cytometer (BD Biosciences) and data analyzed by the Flowing software.

Poursafavi Z., Abroun S., Kaviani S, & Roodbari N.H. (2022). Differentiation of Alginate-Encapsulated Wharton Jelly-Derived Mesenchymal Stem Cells into Insulin Producing Cells. Cell Journal (Yakhteh), 24(8), 449-457.

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Isolation and Characterization of Bone Marrow Mesenchymal Stem Cells

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BMSCs obtained from Cyagen Biotechnology Co., Ltd. (Suzhou, China) (https://www.cyagen.com/cn/zh-cn/product/bone-marrow-msc-MUBMX-01001.html)
were cultured in DMEM supplemented with 10% FBS and maintained at 37°C in a
saturated humidity atmosphere containing 95% air and 5% CO₂. Flow
cytometry analysis showed that BMSCs CD29 (ab21845, Abcam), CD44 (ab21024,
Abcam), and CD73 (ab239246, Abcam) were positive, while hematopoietic markers
CD34 (ab18224, Abcam), CD45 (ab27287, Abcam), and HLA-DR (ab1182, Abcam) were
negative.

Li G.Q., Fang Y.X., Liu Y., Meng F.R., Wu X., Zhang C.W., Zhang Y., Liu Y.Q, & Liu D. (2021). MicroRNA-21 from bone marrow mesenchymal stem cell-derived extracellular vesicles targets TET1 to suppress KLF4 and alleviate rheumatoid arthritis. Therapeutic Advances in Chronic Disease, 12, 20406223211007369.

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Multiparameter Flow Cytometric Analysis

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All antibodies were obtained from commercial sources. Staining for CD45 [FITC conjugated mouse monoclonal (B-A11), Abcam, ab27287] and CD71 [AlexaFluor647 conjugated mouse monoclonal (MEM75), Abcam, ab187777] was performed on ice for 30 min each. Additional WT-1 (1/50 dilution; rabbit anti-human, Abcam, ab15249) staining was performed on ice for 30 min after permeabilization with 0.1% Tween-PBS and blocking with 10% goat serum in FACS buffer. For flow cytometry, AlexaFluor405 goat anti-rabbit antibody (1/2,000 dilution; Abcam, ab175655) was used as a secondary antibody. For fluorescence histochemistry, FITC goat anti-rabbit antibody (1/100 dilution; Thermo Fischer, F-2765) was used as a secondary antibody to allow for Hoechst staining of the sections.

Kienzle A., Servais A.B., Ysasi A.B., Gibney B.C., Valenzuela C.D., Wagner W.L., Ackermann M, & Mentzer S.J. (2018). Free-Floating Mesothelial Cells in Pleural Fluid After Lung Surgery. Frontiers in Medicine, 5, 89.

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BMSC Immunophenotyping by Flow Cytometry

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BMSCs were digested with 0.25% trypsin to collect 4 × 10⁵ cells, centrifuged at 1000 rpm for 4 min at room temperature. After discarding the supernatant, single cell suspension was prepared. Afterwards, BMSCs were incubated with monoclonal antibodies of cell surface antigens containing CD29-PE (ab218273, Abcam, USA), CD44-PE (ab23396), CD90-APC (ab25322), CD105-PE/Cy7 (ab272352), CD146-FITC (ab78451), and CD45-FITC (ab27287) on the ice in the dark for 30 min. After centrifuging at 1000 rpm for 5 min, the supernatant was discarded. Afterwards, the cells were resuspended in 500 μL PBS and examined utilizing a FACSCalibur flow cytometer (BD Biosciences, USA).

Liang Y., Zhou R., Liu X., You L., Chen C., Ye X., Wei W., Liu J., Dai J., Li K, & Zhao X. (2022). Leukemia Inhibitory Factor Facilitates Self-Renewal and Differentiation and Attenuates Oxidative Stress of BMSCs by Activating PI3K/AKT Signaling. Oxidative Medicine and Cellular Longevity, 2022, 5772509.

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Mouse Bone Marrow Stromal Cell Culture

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The mouse BMSCs were purchased from the Cyagen Biosciences Inc. (Suzhou, Jiangsu, China) (https://www.cyagen.com/cn/zh-cn/product/bone-marrow-msc-MUBMX-01001.html) and cultured in H-DMEM (Solarbio Co., Ltd., Beijing, China) with 10% FBS at 37°C with 5% CO₂ under saturated humidity.
The contents of related markers of BMSCs were detected by flow cytometry. The cells were found to be positive for BMSC markers, including CD29 (ab21845, Abcam), CD44 (ab25024, Abcam), and CD73 (ab239246, Abcam) while negative for hematopoietic markers, including CD34 (ab18224, Abcam), CD45 (ab27287, Abcam), and human leukocyte antigen-DR (HLA-DR) (ab1182, Abcam).

Jiang Y., Zhang J., Li Z, & Jia G. (2020). Bone Marrow Mesenchymal Stem Cell-Derived Exosomal miR-25 Regulates the Ubiquitination and Degradation of Runx2 by SMURF1 to Promote Fracture Healing in Mice. Frontiers in Medicine, 7, 577578.

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Characterization of Human BMSCs

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Human BMSCs were incubated in the DMEM-F12 supplement with 10% FBS, L-glutamine, and penicillin-streptomycin solution (both 100x, diluted to 1x for use). The fresh medium was applied to wash off the nonadherent cells after 48 h incubation. The cells of passage 3 with 80% confluence were chosen for BMSC identification.
Human BMSCs were centrifuged after 0.25% trypsin detaching. The precipitates obtained from centrifugation were washed twice with 1x PBS to count the cells. Human BMSCs (1 × 10⁶ cells/ml) were probed with CD45 antibody (ab27287, Abcam, Cambridge, MA, USA), CD105 antibody (ab155367, Abcam), and CD73 antibody (ab157335, Abcam) and subsequently analyzed using flow cytometry.

Zhao H, & He Y. (2021). Lysophosphatidylcholine Offsets the Protective Effects of Bone Marrow Mesenchymal Stem Cells on Inflammatory Response and Oxidative Stress Injury of Retinal Endothelial Cells via TLR4/NF-κB Signaling. Journal of Immunology Research, 2021, 2389029.

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