Cd34 biotin

Manufactured by Thermo Fisher Scientific

Sourced in United States

CD34-biotin is a reagent used in flow cytometry applications. It is a biotin-conjugated monoclonal antibody that specifically binds to the CD34 antigen, which is expressed on the surface of hematopoietic stem and progenitor cells. This reagent can be used to identify and isolate CD34-positive cells from various sample types.

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Lab products found in correlation

Facsaria, by BD (6 mentions) Streptavidin apc, by BD (6 mentions) Cd31 biotin, by Thermo Fisher Scientific (2 mentions) Cd45 biotin, by Thermo Fisher Scientific (2 mentions) Streptavidin apc cy7, by Thermo Fisher Scientific (2 mentions) Cd24 pe, by Thermo Fisher Scientific (2 mentions) Sca 1 apc, by Thermo Fisher Scientific (2 mentions) α6 integrin buv395, by BD (2 mentions) Sca1 bv421, by BD (2 mentions) Propidium iodide, by Merck Group (2 mentions) α6 integrin pacific blue, by BD (2 mentions) Pdgfrα apc, by BioLegend (2 mentions) Cd31 pe cy7, by BioLegend (2 mentions) Cd45 fitc, by BioLegend (2 mentions) 7 amino actinomycin d staining, by BD (1 mentions) Epcam pe cy7, by Thermo Fisher Scientific (1 mentions) Pe annexin 5 apoptosis detection kit with 7 aad, by BioLegend (1 mentions) Anti α6 integrin pe, by BD (1 mentions) Facsdiva software, by BD (1 mentions) Dispase 2, by STEMCELL (1 mentions)

Spelling variants of this product

Biotin-conjugated antibodies to CD34 (3 citations) Anti-CD34-Biotin (2 citations) Biotin-conjugated anti-CD34 (2 citations)

10 protocols using cd34 biotin

Lung Cell Isolation and FACS Analysis

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Lung cell suspensions were prepared using an elastase digestion and stained for fluorescence-activated cell sorting (FACS), as previously described (20 (link)). Briefly, cells were resuspended in Hanks' balanced saline solution buffer supplemented with 2% fetal bovine serum, 0.1 mM EDTA, 10 mM HEPES, 100 IU/ml penicillin and 100 µg/ml streptomycin (HBSS⁺). Cells were then stained with the primary antibodies on ice for 45 min. The following antibodies were employed: EpCAM-PE-Cy7 (25-5791-80, 1:100), CD31-Biotin (13-0311-81, 1:40), CD34-Biotin (13-0341-81, 1:10), CD45-Biotin (13-0451-81, 1:100), Sca-1-APC (17-5981-81, 1:100), and CD24-PE (12-0242-81, 1:20) (all from eBioscience, Inc., San Diego, CA, USA). Cells were subsequently stained with the secondary antibody on ice for 40 min using streptavidin-APC-Cy7 (47-4317-82, 1:100; eBioscience, Inc.). Dead cells were identified using 7-aminoactinomycin D staining (BD Biosciences, Franlkin Lakes, NJ, USA).

Li X., Yang L., Sun X., Wu J., Li Y., Zhang Q., Zhang Y., Li K., Wu Q, & Chen H. (2017). The role of TGFβ-HGF-Smad4 axis in regulating the proliferation of mouse airway progenitor cells. Molecular Medicine Reports, 16(6), 8155-8163.

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Isolation and Characterization of Murine Epidermal Stem Cells

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Mouse tail skin was incubated in 0.25% Trypsin/EDTA overnight at 4°C and for 30 min at 37°C. Single cell solution was prepared by scraping and subsequent filtering with strainers (70 μm, followed by 40μm). Cells were stained with the following antibodies for 30 min on ice: CD34-biotin (1:50, eBioscience, 13–0341), Streptavidin-APC (1:100, BD Biosciences, 554067), α6-integrin-BUV395 (1:100, BD Biosciences, custom order) and Sca1-BV421 (1:100, BD Biosciences, 562729). The dead cells were excluded by using propidium iodide (P4864, Sigma Aldrich). FACS (Fluorescence-activated cell sorting) analyses were performed with FACS Aria (BD Biosciences). The data was processed with FlowJo software to measure the mean fluorescence intensity of basal markers. For the detection of apoptotic cells, cells were stained with PE Annexin V Apoptosis Detection Kit with 7-AAD (BioLegend) according to the manufacturer’s instructions.

Changarathil G., Ramirez K., Isoda H., Sada A, & Yanagisawa H. (2019). Wild-type and SAMP8 mice show age-dependent changes in distinct stem cell compartments of the interfollicular epidermis. PLoS ONE, 14(5), e0215908.

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Isolation and Analysis of Mouse Skin Cells

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Mouse back skin was incubated in 0.25% trypsin/versene overnight at 4°C and for 30 min at 37°C. Single cell suspensions were prepared by scraping off the fat and subcutaneous tissue from the dermal side of the skin followed by enzymatic digestions and subsequent filtering with strainers (70 mm, followed by 40 mm). Cells were stained with the following antibodies for 30 min on ice: CD34-biotin (1:50, eBioscience), Streptavidin-APC (1:100, BD Biosciences) and α6-integrin-Pacific blue (1:100, BD Biosciences, custom order). Dead cells were excluded by propidium iodide (PI) (Sigma) staining. FACS (FACS Aria, BD Biosciences) were performed in the Cornell Flow Cytometry facility. FACS data were analyzed with the FlowJo software.

Sada A., Jacob F., Leung E., Wang S., White B.S., Shalloway D, & Tumbar T. (2016). Defining the cellular lineage hierarchy in the inter-follicular epidermis of adult skin. Nature cell biology, 18(6), 619-631.

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Isolation and Characterization of Hair Follicle Stem Cells

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Dorsal skin of wild type control littermate and K14-sPLA₂-IIA homozygous mice at PD21 and PD49 was harvested and scrapped for fat removal, followed by overnight incubation in 0.25% trypsin at 4 °C. FACS experiments were performed as described previously^{7 (link)}. Single cell suspension was obtained by first passing through 70 μm and then 40 μm cell strainers (BD Biosciences). Cells were stained with trypan blue and the hematocytometer chamber was used to count the cells. Further cells were stained by using the hair follicle stem cells markers: CD34-Biotin (eBiosciences), Streptavidin-APC (BD Pharmingen), and anti–α6-integrin-PE (BD Pharmingen). After washing cells were subjected to FACS acquisition using a FACS Aria and data was analyzed by using FACS DiVa software (BD Biosciences).

Chovatiya G.L., Sarate R.M., Sunkara R.R., Gawas N.P., Kala V, & Waghmare S.K. (2017). Secretory phospholipase A2-IIA overexpressing mice exhibit cyclic alopecia mediated through aberrant hair shaft differentiation and impaired wound healing response. Scientific Reports, 7, 11619.

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Isolating Mouse Skin Cells

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Mouse back skin was minced and dissociated into single-cell suspension at 37°C for a total of two hours in the following enzymatic mixture: 2 mg/ml collagenase type I (Worthington), 1.5 mg/ml collagenase type II (Worthington), 2.5 mM Ca⁺², 1 mM Mg⁺² and 1% BSA in 1x Hank’s Balanced Salt Solution (HBSS). In addition, 1U/ml Dispase II (Stemcell Technologies) and 50 U/ml DNase I (Worthington) were added in the above mixture for the final 1 and 0.5 hours respectively. After two hours, enzymes activity was neutralized by the addition of serum-containing medium, followed by serial filtration through 70 and 40-micron strainers. The cell suspension was washed with 1x-PBS containing 5% FBS and sequentially stained with CD34-biotin (1:50, eBioscience) and Streptavidin-APC (1:100, BD Biosciences) antibodies for 30 minutes each, on ice. LIVE/DEAD™ Fixable Aqua Dead Cell Stain Kit (ThermoFisher) was used to label the dead cells. FACS (FACS Aria, BD Biosciences) was performed in the Cornell Flow Cytometry facility. FACS data were analyzed with the FlowJo (FlowJo™ Software, v10.5.0, BD Biosciences).

Chovatiya G., Ghuwalewala S., Walter L.D., Cosgrove B.D, & Tumbar T. (2021). High resolution single cell transcriptomics reveals heterogeneity of self-renewing hair follicle stem cells. Experimental dermatology, 30(4), 457-471.

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Tumor Cell and Immune Cell Sorting

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TIC and immune cell populations were harvested from the same mice for each experiment. Tumor cell sub-populations were sorted by flow cytometry using CD34-biotin (eBioscience), CD45-FITC, PDGFRα-APC and CD31-PECy7 (Biolegend) antibodies followed by streptavidin-PE. TAMs were isolated using CD45-FITC, CD11b-PECy7 (Biolegend), and F4/80-biotin, followed by streptavidin-PE.

Tham M., Khoo K., Yeo K.P., Kato M., Prevost-Blondel A., Angeli V, & Abastado J.P. (2015). Macrophage depletion reduces postsurgical tumor recurrence and metastatic growth in a spontaneous murine model of melanoma. Oncotarget, 6(26), 22857-22868.

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Isolation and Characterization of Mouse Lung Cell Types

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A single-cell suspension was prepared by digestion of lung tissues from 6–10-week-old C57BL/6 mice with elastase and DNase I. Freshly isolated cells were suspended in Hanks balanced saline
solution (Solarbio, Beijing, China) with 2% fetal bovine serum (FBS, Gibco), 0.01% Penicillin- streptomycin (Gibco, USA) and 10 mM HEPES (Sigma). The primary antibodies used to stain cells
included CD24-PE (1:25, eBioscience, San Diego, CA, USA), EpCAM-PE-Cy7 (1:50, Biolegend, San Diego, CA, USA), CD31-biotin (1:50, eBioscience), CD45-biotin (1:100, eBioscience), CD34-biotin
(1:16, eBioscience), and Sca-1-APC (1:100, eBioscience). Then, the secondary antibody Streptavidin-APC-Cy7 (1:100, eBioscience) was used, followed by the addition of 7-amino-actinomycin D
(7-AAD, 1:20, eBioscience) to discriminate the dead cells. Mouse club and AT2 cells were sorted based on their surface expression pattern,
CD31⁻CD34⁻CD45⁻EpCAM⁺CD24⁺Sca-1⁺ and
CD31⁻CD34⁻CD45⁻EpCAM⁺CD24⁻Sca-1⁻, respectively.

Zhao F., Wang J., Wang Q., Hou Z., Zhang Y., Li X., Wu Q, & Chen H. (2022). Organoid technology and lung injury mouse models evaluating effects of hydroxychloroquine on lung epithelial regeneration. Experimental Animals, 71(3), 316-328.

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Isolation of Mouse Skin Stem Cells

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Mouse dorsal and ventral skin were dissected and the subcutaneous and fat tissues were removed from the dermal side of the skin. The skin was incubated in 0.25% trypsin/versene overnight at 4°C and for 30 min at 37°C. The single‐cell solution was prepared by scraping the epidermis and subsequent filtering with strainers (70 μm, followed by 40 μm). Cells were stained with the following antibodies for 30 min on ice: CD34‐biotin (1:50, eBioscience), Streptavidin‐APC (1:100, BD Biosciences), α6‐integrin‐BUV395 (1:100, BD Biosciences, custom order) and Sca1‐BV421 (1:100, BD Biosciences). The dead cells were excluded by staining with propidium iodide (Sigma‐Aldrich). Cell isolation was performed with FACS Aria (BD Biosciences), and the data were analyzed with the FlowJo software (BD, Franklin Lakes, NJ).

Oinam L., Changarathil G., Raja E., Ngo Y.X., Tateno H., Sada A, & Yanagisawa H. (2020). Glycome profiling by lectin microarray reveals dynamic glycan alterations during epidermal stem cell aging. Aging Cell, 19(8), e13190.

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Isolation and Analysis of Mouse Skin Cells

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Immune Cell Profiling in Mouse Tumors

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TIC and immune cell populations were harvested from the same mice for each experiment. CD45⁺ immune cells were depleted from the tumor with anti-CD45-PE antibody and the anti-PE EasySep selection kit (StemCell). Tumor cell sub-populations were sorted by flow cytometry using CD34-biotin (eBioscience), anti-CD271 (Millipore) and CD45-FITC (Biolegend) antibodies followed by streptavidin-PE and anti-rabbit PECy7 (Invitrogen) antibodies. Immune cell population were sorted using CD45, CD3, CD4, CD8, NK1.1, CD19, CD11b (Biolegend), and CD68 (AbDserotec) conjugated to appropriate fluorophores. Stromal cell populations were sorted using CD31-PECy7 and PDGFRα-APC (Biolegend).

Tham M., Tan K.W., Keeble J., Wang X., Hubert S., Barron L., Tan N.S., Kato M., Prevost-Blondel A., Angeli V, & Abastado J.P. (2014). Melanoma-initiating cells exploit M2 macrophage TGFβ and arginase pathway for survival and proliferation. Oncotarget, 5(23), 12027-12042.

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