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Anti brdu fitc

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Anti-BrdU-FITC is a fluorescent-labeled antibody that binds to bromodeoxyuridine (BrdU), a synthetic nucleoside analog of thymidine. This product is used in flow cytometric analysis to detect and quantify cells that have incorporated BrdU during DNA synthesis, which is indicative of cell proliferation.

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Lab products found in correlation

Bromodeoxyuridine (brdu), by Merck Group (3 mentions) Iscove modified dulbecco media (imdm), by Thermo Fisher Scientific (2 mentions) Anti chk1 sc 8408, by Santa Cruz Biotechnology (1 mentions) Anti p38 mapk t180 y182, by Cell Signaling Technology (1 mentions) Anti p44 42 mapk erk1 2 t202 y204, by Cell Signaling Technology (1 mentions) Anti sapk jnk t183 y185, by Cell Signaling Technology (1 mentions) Anti α tubulin sc 23948, by Santa Cruz Biotechnology (1 mentions) Anti mcm2 sc 9839, by Santa Cruz Biotechnology (1 mentions) Anti brdu ab6326, by Abcam (1 mentions) Anti mcm2 s53 a300 756a, by Fortis Life Sciences (1 mentions) Cleaved caspase 3, by Cell Signaling Technology (1 mentions) Anti sapk jnk, by Cell Signaling Technology (1 mentions) Anti p44 42 mapk erk1 2, by Cell Signaling Technology (1 mentions) P38 mapk, by Cell Signaling Technology (1 mentions) Goat anti mouse igg, by Jackson ImmunoResearch (1 mentions) Anti brdu, by BD (1 mentions) Anti phospho h2ax s139, by Merck Group (1 mentions) Caspase 9, by Cell Signaling Technology (1 mentions) Mcl 1, by Cell Signaling Technology (1 mentions) Alexa fluor 488 anti mouse igg, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

FITC-conjugated anti-BrdU antibody (8 citations) Anti-BrdU (6 citations) Anti-BrdU-FITC antibody (5 citations)

7 protocols using anti brdu fitc

1

Bone Marrow Progenitor Cell Proliferation

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Bone marrow cells were isolated from WD-fed WT or Apoe−/− mice treated with vehicle or TG101348 (240 mg/day) for 6 weeks and then were cultured in IMDM (Invitrogen) with 10% FBS and 1 μM BrdU (Sigma). After 12-h incubation, cells were stained as described above (lineage changed to APC-conjugated and CD34 changed to PE-conjugated antibodies) for progenitor cells together with FITC-anti BrdU (Biolegend). Proliferation was quantified as percentage of BrdU + cells by flow cytometry.
Tang Y., Liu W., Wang W., Fidler T., Woods B., Levine R.L., Tall A.R, & Wang N. (2020). Inhibition of JAK2 Suppresses Myelopoiesis and Atherosclerosis in Apoe−/− Mice. Cardiovascular Drugs and Therapy, 34(2), 145-152.
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2

Antibody Characterization for Cell Signaling Assays

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Antibodies used in this study are as follows. Anti-human Claspin was generated against the human recombinant Claspin with aa896–1,014 produced in E. coli. Anti-Chk1 phospho-S345 (#2348), anti-Chk1 phospho-S317 (#2344), anti-p44/42 MAPK (Erk1/2) (#4695), anti- SAPK/JNK (#9252), p38 MAPK (#8690), anti-p38 MAPK T180/Y182 (#4511), anti-p44/42 MAPK (Erk1/2) T202/Y204 (#4370), anti-SAPK/JNK T183/Y185 (#4668), Caspase-9 (#9508), Cleaved Caspase-3(#9661), and Mcl-1 (#5453) were obtained from Cell Signaling. Anti-α Tubulin (sc23948), anti-MCM2 (sc-9839), and anti-Chk1 (sc-8408) were obtained from Santa Cruz. Anti-phospho-H2A.X S139 (06-536) was purchased from Merck. Anti-BrdU (Ab6326) was purchased from Abcam. Anti-ATR phospho-T1989 (GTX128145) was purchased from GeneTex. Anti-BrdU (555627) was purchased from BD Pharmingen. Anti-H2A.X phospho-S139 (613402), anti-Rat IgG Alexa Fluor 555 (405420), and FITC-Anti-BrdU (364104) were purchased from Biolegend. RPA32 phospho-S4/S8 (A300-245A) and anti-MCM2 S53(A300-756A) were purchased from Bethyl. Anti-Mouse IgG Alexa Fluor 488 (A-11017) was purchased from Invitrogen. Goat Anti-Rabbit IgG HRP (111-035-003) and Goat Anti-Mouse IgG (115-035-003) were purchased from Jackson ImmunoResearch Laboratory.
Hsiao H.W., Yang C.C, & Masai H. (2023). Claspin-Dependent and -Independent Chk1 Activation by a Panel of Biological Stresses. Biomolecules, 13(1), 125.
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3

Analyzing Bone Marrow Progenitor Proliferation

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Bone marrow cells were isolated from WD-fed WT or Apoe−/− mice treated with vehicle or TG101348 (240 mg/day) for 6 weeks and then were cultured in IMDM(Invitrogen) with 10% FBS and 1μM BrdU (Sigma). After 12 hours incubation, cells were stained as described above (Lineage changed to APC conjugated and CD34 changed to PE conjugated antibodies) for progenitor cells together with FITC-anti BrdU (Biolegend). Proliferation was quantified as percentage of BrdU+ cells by flow cytometry.
Tang Y., Liu W., Wang W., Fidler T., Woods B., Levine R.L., Tall A.R, & Wang N. (2020). Inhibition of JAK2 Suppresses Myelopoiesis and Atherosclerosis in Apoe−/− Mice. Cardiovascular Drugs and Therapy, 34(2), 145-152.
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4

Multiparameter Flow Cytometry Staining

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Cell staining was performed at 4°C with Fc Block (hybridoma 2.4G2) in FACS staining buffer (0.5% FBS and 0.1% NaN3 in dPBS). Anti–B220-FITC, anti–B220-PE, anti–CD21-APC, Annexin V-FITC, anti–BrdU-FITC, and Streptavidin-PE were purchased from BioLegend. Anti–CD23-biotin and anti–CD138-biotin were purchased from BD. Anti–CD69-biotin, anti–MHC-II-PE, and anti–TLR9-FITC were purchased from eBioscience. Flow data were acquired by the FACSCanto II (BD) and analyzed by FlowJo software.
Chen T.T., Tsai M.H., Kung J.T., Lin K.I., Decker T, & Lee C.K. (2016). STAT1 regulates marginal zone B cell differentiation in response to inflammation and infection with blood-borne bacteria. The Journal of Experimental Medicine, 213(13), 3025-3039.
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5

Cell Cycle Analysis by BrdU-PI Staining

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To analyze cell cycle distribution, bromodeoxyuridine (BrdU) – propidium iodide (PI) staining was performed. Briefly, cells were plated at 7.5 × 104 cells per 6-well plate (7800/cm2), and allowed to attach overnight. Once attached, media was replaced and cells were then treated with vehicle control (DMSO) or RAGE inhibitors at 1 µM for 48 h. During the final hour of culture, cells were pulsed with 10 µM BrdU (BioLegend Catalog# 423401), collected by trypsinization, fixed in iced-cold 70% ethanol, and stored at −20C for at least 2 h. Fixed cells were labeled with anti-BrdU-FITC (BioLegend Catalog# 364104) following manufacturer’s instructions, and counterstained with propidium iodide (p4170-25MG Sigma). Labeled cells were analyzed by flow cytometry (BD Fortessa), and results analyzed using FlowJo software.
Magna M., Hwang G.H., McIntosh A., Drews-Elger K., Takabatake M., Ikeda A., Mera B.J., Kwak T., Miller P., Lippman M.E, & Hudson B.I. (2023). RAGE inhibitor TTP488 (Azeliragon) suppresses metastasis in triple-negative breast cancer. NPJ Breast Cancer, 9, 59.
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6

Cell Cycle Analysis by BrdU and PI

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Cells were seeded in 60 mm dishes (Corning 430166) (75,000 cells/well) and treated with vehicle or drugs 24 h later. Following the indicated drug exposure times, the cells were pulsed with 10 μM bromodeoxyuridine (BrdU; Sigma) for 90 min, trypsinized, washed with 1x PBS, fixed in ice-cold 70% ethanol, denatured with 2 N HCl for 25 min at RT, followed by inactivation with 0.1 M sodium borate, pH 8.5. Cells were then incubated with anti-BrdU FITC (Biolegend, Clone 3D4, 364104) (5 μl in 100 μl cells suspended in 0.2% BSA in PBS) for 20 min at RT, spun down then resuspended in 500 μl PI/RNase staining buffer (BD Biosciences, 550825) for 15 min both in the dark. Cell cycle analysis was performed on a FACSCalibur cytometer/CellQuest Pro.
El Touny L.H., Hose C., Connelly J., Harris E., Monks A., Dull A.B., Wilsker D.F., Hollingshead M.G., Gottholm-Ahalt M., Alcoser S.Y., Mullendore M.E., Parchment R.E., Doroshow J.H., Teicher B.A, & Rapisarda A. (2021). ATR inhibition reverses the resistance of homologous recombination deficient MGMTlow/MMRproficient cancer cells to temozolomide. Oncotarget, 12(21), 2114-2130.
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7

Quantifying Neutrophil Proliferation In Vivo

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Mice were injected (i.p.) with 2.5 mg of BrdU (Thermo Fisher, #000103) (Figure S3). Neutrophils containing BrdU that were present in the mouse PB and lung samples were detected using an anti-BrdU antibody. Briefly, single-cell suspensions from PB and lung tissues were first stained for cell surface antigens (CD45-APC/Cy7, CD11b-PE/Cy7, Ly-6G-APC, and CD62L-PE), and then fixed and permeabilized using a BrdU Flow Kit (BD Bioscience, #557891). Cells were treated with DNase I (Sigma-Aldrich, 100 μg/mL, #D5025) for 1 h at 37°C to expose BrdU incorporated in DNA. Then, cells were stained for BrdU with fluorescent antibodies (anti-BrdU-FITC, Biolegend, #364103).
Wang Z., Yang C., Li L., Zhang Z., Pan J., Su K., Chen W., Li J., Qiu F, & Huang J. (2020). CD62Ldim Neutrophils Specifically Migrate to the Lung and Participate in the Formation of the Pre-Metastatic Niche of Breast Cancer. Frontiers in Oncology, 10, 540484.
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