Esa fitc

Manufactured by BD

Sourced in United States

The ESA-FITC is a fluorescent antibody used in flow cytometry analysis. It binds to the extracellular signal-regulated kinase (ESA) protein and emits fluorescence when excited by a light source, allowing for the detection and quantification of ESA-positive cells.

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Lab products found in correlation

Cd44 apc, by BD (4 mentions) Cd24 pe, by BD (3 mentions) Typsin edta, by Thermo Fisher Scientific (1 mentions) Cd44 pe, by BD (1 mentions) Alexa fluor 488 goat anti rabbit, by Thermo Fisher Scientific (1 mentions) Cd24 fitc, by BD (1 mentions) Cd49f apc, by BD (1 mentions) Cd24 pe antibody, by BD (1 mentions) Facscanto flow cytometer, by BD (1 mentions) Cd133 apc, by BioLegend (1 mentions) Aldefluor assay, by STEMCELL (1 mentions)

Close spelling variants of this product

Anti-ESA-FITC (2 citations)

6 protocols using esa fitc

Isolation of CD44+CD24-/low breast cancer stem cells

Cited 2 times

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MCF-7 and T47D cells were cultured in DMEM-H (HyClone, USA) containing 10% fetal bovine serum (FBS) at 37 °C in a 5% CO₂ incubator. Cells in the logarithmic growth phase, approximately 70–80% confluent, were harvested and digested into a single cell suspension by trypsin digestion.
The complete MammoCult™ medium containing 5% MammoCult™ proliferation supplement, 4 μg/mL heparin, and 0.48 μg/mL hydrocortisone was used. MCF-7 cells were resuspended in complete MammoCult™ medium, and T47D cells resuspended in complete DMEM/F-12 medium. The cell suspension (4 × 10³) was seeded into a 6-well plate and cultured at 37 °C and 5% CO₂. After 7 days of culture, the spheres were collected and centrifuged at 350 g for 5 min, and the supernatant was discarded. Then, 1 mL of Accutase cell dispersion solution was added to digest the spheres, followed by adding 9 mL of sterile PBS solution and centrifugation at 350 g for 5 min. Finally, the supernatant was discarded, and the cells were collected.
CD44⁺CD24^−/low BCSCs were isolated from MCF-7 and T47D cells by staining with CD44-APC, CD24-PE and ESA-FITC (BD Pharmingen, USA) antibodies via FACS as described in our previous research [8 (link), 20 (link)].

Xia L., Li F., Qiu J., Feng Z., Xu Z., Chen Z, & Sun J. (2020). Oncogenic miR-20b-5p contributes to malignant behaviors of breast cancer stem cells by bidirectionally regulating CCND1 and E2F1. BMC Cancer, 20, 949.

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Isolation of MUC1-ESA+ Subpopulation

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When MCF-10A cell confluence reached about 80 %, single cells were obtained by 0.25 % typsin/EDTA (Gibco, USA) digestion, stained by MUC1-PE (BD Pharmingen, USA) and ESA-FITC (BD Pharmingen, USA). MUC1⁻ESA⁺ subpopulation was sorted by fluorescence-activated cell sorting (FACS, MoFlo, Dako-Cytomation, USA). The rest proportion of MCF-10A cells excluding MUC1⁻ESA⁺ was also sorted as control counterparts. Through FACS sorting, MUC1⁻ESA⁺ subpopulation was highly purified (purity greater than 98 %).

Feng Z.M., Qiu J., Chen X.W., Liao R.X., Liao X.Y., Zhang L.P., Chen X., Li Y., Chen Z.T, & Sun J.G. (2015). Essential role of miR-200c in regulating self-renewal of breast cancer stem cells and their counterparts of mammary epithelium. BMC Cancer, 15, 645.

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Immunofluorescence Analysis of Paraffin-Embedded Xenograft Tumors

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The paraffin-embedded PK45 xenograft tumor sections were deparaffinization by xylene and rehydration by 100%, 95% ethanol and sections rinsed with dH2O. Then the slides were boiled in 10 mM sodium citrate buffer pH 6.0 for antigen unmasking. The block specimen was blocked in a blocking buffer (1X PBS/5% normal serum/0.3% Triton™ X-100) for 1h, after which diluted fluorochrome-conjugated primary antibody CD44-PE (BD Pharmingen™) and ESA-FITC (BD Pharmingen™) were applied and the specimen was incubated overnight at 4°C. After rinsing three times in 1X PBS for 5 min each, coversliped slides with Prolong® Gold Antifade Reagent (#9071) or Prolong® Gold Antifade Reagent with DAPI (#8961). Ten fields were selected and expression was evaluated in 10 fields with high power (x200) microscopy [39 (link)].

Sai S., Wakai T., Vares G., Yamada S., Kamijo T., Kamada T, & Shirai T. (2015). Combination of carbon ion beam and gemcitabine causes irreparable DNA damage and death of radioresistant pancreatic cancer stem-like cells in vitro and in vivo. Oncotarget, 6(8), 5517-5535.

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Isolation and Analysis of Mammary Stem Cells

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To analyze mammary stem and progenitor populations, Lin⁻ cells isolated from mouse MECs were incubated with CD24-FITC, CD49f-APC, and/or CD61-FITC antibodies (BD Biosciences) on ice for 15 min. To measure the breast CSC population, cells were labeled with ESA-FITC, CD44-APC, and CD24-PE antibodies (BD Biosciences) at 4°C for 15 min. The analysis was performed using a FACSCanto Flow Cytometer (BD Biosciences). To measure basal and luminal marker expression, cells were stained with primary antibodies against basal and luminal markers, incubated with goat anti-mouse Cy5 (Invitrogen) for luminal markers or goat anti-rabbit Alexa Fluor 488 (Invitrogen) for basal markers, and analyzed by flow cytometry.

Shin D.H., Park J.H., Lee J.Y., Won H.Y., Jang K.S., Min K.W., Jang S.H., Woo J.K., Oh S.H, & Kong G. (2015). Overexpression of Id1 in transgenic mice promotes mammary basal stem cell activity and breast tumorigenesis. Oncotarget, 6(19), 17276-17290.

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Identifying Cancer Stem Cells via Aldefluor

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The ALDEFLUOR assay (Stemcell Technologies, Durham, NC) was performed according to the manufacturer's guidelines to identify cells with high ALDH activity. Cells were passed through a 35-µm filter, suspended in ALDEFLUOR assay buffer + BODIPY-aminoacetaldehyde (BAAA) and incubated for 45 min at 37°C in the presence or absence of the ALDH inhibitor diethylaminobenzaldehyde (DEAB). CD44-APC (BD Biosciences), PROCR-PE (BD Biosciences), ESA-FITC (BD Biosciences), CD24-PE (BD Biosciences) and CD133-APC (Biolegend) antibodies were incubated with single cells in PBS/1% FBS for 30 min at 4°C. Cells were stained with propidium iodide (PI) or 7-AAD to exclude non-viable cells. For experiments using cells transfected with GFP-tagged hNICD, CD44-APC was used to detect the CSCs. GFP analysis by FACS analysis was used to verify transfection of the hNICD construct. The CSC markers representing tumor-initiating populations in the cell lines used and their respective references are in the Table 1.

Bhola N.E., Jansen V.M., Koch J.P., Li H., Formisano L., Williams J.A., Grandis J.R, & Arteaga C.L. (2015). Treatment of Triple Negative Breast Cancer With TORC1/2 Inhibitors Sustains a Drug-resistant and Notch-dependent Cancer Stem Cell Population. Cancer research, 76(2), 440-452.

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Isolation and Culture of Breast Cancer Stem Cells

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Induction culture of BCSCs MCF-7 and T47D cells were cultured in DMEM-H (HyClone, USA) containing 10% fetal bovine serum (FBS) at 37 °C in a 5% CO 2 incubator. Cells in the logarithmic growth phase, approximately 70-80% confluent, were harvested and digested into a single cell suspension by trypsin digestion.
The complete MammoCult™ medium containing 5% MammoCult™ proliferation supplement, 4 µg/mL heparin, and 0.48 µg/mL hydrocortisone was used. MCF-7 cells were resuspended in complete MammoCult™ medium, and T47D cells resuspended in complete DMEM/F-12 medium. The cell suspension (4×10 3 ) was seeded into a 6-well plate and cultured at 37 °C and 5% CO 2 . After 7 days of culture, the spheres were collected and centrifuged at 350 g for 5 min, and the supernatant was discarded. Then, 1 mL of Accutase cell dispersion solution was added to digest the spheres, followed by adding 9 mL of sterile PBS solution and centrifugation at 350 g for 5 minutes. Finally, the supernatant was discarded, and the cells were collected.
CD44 + CD24 -/low BCSCs were isolated from MCF-7 and T47D cells by staining with CD44-APC, CD24-PE and ESA-FITC (BD Pharmingen, USA) antibodies via FACS as described in our previous research [8, 20] .

Xia L., Li F., Qiu J., Feng Z., Xu Z., Chen X., Chen Z., & Jian-guo S. (2020). Oncogenic miR-20b-5p contributes to malignant behaviors of breast cancer stem cells by bidirectionally regulating CCND1 and E2F1.

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