Streptavidin pe texas red

Manufactured by Thermo Fisher Scientific

Streptavidin PE-Texas red is a fluorescent conjugate that combines streptavidin with the PE-Texas Red dye. Streptavidin is a tetrameric protein that binds strongly to biotin, a common label used in various biological applications. The PE-Texas Red dye provides a fluorescent signal that can be detected and quantified using appropriate instrumentation.

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Texas Red (171 citations) Streptavidin-PE (135 citations) PE Texas Red (7 citations) Streptavidin Texas red (6 citations)

4 protocols using streptavidin pe texas red

Mixed Bone Marrow Chimeras for Cell Fate Analysis

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Mixed bone marrow chimeras were generated by transplanting a 1:3 ratio of Ly5.1⁺ competitor cells and Ly5.2⁺ test cells (5 × 10⁶ total bone marrow cells) into lethally irradiated (1100 rad) Ly5.1⁺ Ly5.2⁺ recipients. Test bone marrow cells were isolated from wild-type, Bak^+/Q75L, or Bak^Q75L/Q75L mice. Bone marrow chimeras were analyzed 12 wk after reconstitution by flow cytometry performed with a Fortessa (BD Biosciences), and data were analyzed using FlowJo software (Tree Star). Monoclonal antibodies were produced at the Walter and Eliza Hall Institute unless otherwise indicated and labeled in-house with fluorescein isothiocyanate, R-phycoerythrin, R-phycoerythrin-Cy7, allophycocyanin, Alexa700, or biotin. Clone numbers were CD4 (GK1.5), CD8 (53.6.7), CD19 (1D3), CD44 (IM7), CD45.1 (A20), CD45.2 (S450-15-2), TER119 (Ly76), GR1 (RB6-8C5), CD11b (M1/70), Sca1 (D7), Kit (ACK4), and B220 (RA3-6B2). CD62L-allophycocyanin was purchased from eBioscience. Streptavidin PE-Texas red was purchased from Life Technologies. Fluoro-Gold (Sigma) was used to exclude dead cells.

Lee E.F., Grabow S., Chappaz S., Dewson G., Hockings C., Kluck R.M., Debrincat M.A., Gray D.H., Witkowski M.T., Evangelista M., Pettikiriarachchi A., Bouillet P., Lane R.M., Czabotar P.E., Colman P.M., Smith B.J., Kile B.T, & Fairlie W.D. (2016). Physiological restraint of Bak by Bcl-xL is essential for cell survival. Genes & Development, 30(10), 1240-1250.

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Multiparametric Flow Cytometry Analysis

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Organ cellularity was determined by counting of cell suspensions from bone marrow, spleen, and thymus with calibrate beads (BD Biosciences). Cells were stained on ice for 30 minutes, washed once, and then incubated with streptavidin–PE–Texas red (Life Technologies). After a washing step, acquisition was performed on an LSR Fortessa 1 (BD Biosciences) and analyzed with FlowJo (Tree Star). Antibodies for flow cytometry either were labeled in house with FITC, PE, PECy7, APC, or Alexa Fluor 700 or were biotinylated. CD4 (GK1.5), CD8 (53.6.7), CD16/32 (24G2), CD19 (1D3), CD41 (MWReg30), Ter119 (Ly76), Gr1 (RB6-8C5), CD11b (M1/70), Sca1 (D7), Kit (ACK4), B220 (RA3-6B2), and CD34-FITC (RAM34) were purchased from BD Pharmingen, and CD150-PE (TC15-12F12.2) was purchased from BioLegend. Fluorogold (Sigma-Aldrich) was used to exclude dead cells.

Pleines I., Woods J., Chappaz S., Kew V., Foad N., Ballester-Beltrán J., Aurbach K., Lincetto C., Lane R.M., Schevzov G., Alexander W.S., Hilton D.J., Astle W.J., Downes K., Nurden P., Westbury S.K., Mumford A.D., Obaji S.G., Collins P.W., Delerue F., Ittner L.M., Bryce N.S., Holliday M., Lucas C.A., Hardeman E.C., Ouwehand W.H., Gunning P.W., Turro E., Tijssen M.R, & Kile B.T. (2017). Mutations in tropomyosin 4 underlie a rare form of human macrothrombocytopenia. The Journal of Clinical Investigation, 127(3), 814-829.

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Multiparameter Flow Cytometry Panel

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The following monoclonal antibodies and cell dyes were used: CD45.1 FITC, CD45.2 AlexaFluor700, CD45.2 FITC, TCR-beta APC, CD4 PerCP-eFluor710, CD44 APC-eFluor780, CD25 PE-Cy7, L-selectin eFluor450, CD122 biotin (all eBioscience); CD8 Pacific Orange, streptavidin PE-TexasRed, LIVE/DEAD blue (all Invitrogen); and CD45.1 Brilliant Violet 650, CD4 Brilliant Violet 711, and TCR-beta PerCP-Cy5.5 (all BioLegend). Samples were acquired on LSR-II, LSRFortessa, or Fortessa X20 flow cytometers (BD), and analysis was performed with FlowJo software (Treestar).

Rane S., Hogan T., Seddon B, & Yates A.J. (2018). Age is not just a number: Naive T cells increase their ability to persist in the circulation over time. PLoS Biology, 16(4), e2003949.

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Murine Blood and Bone Marrow Analysis

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Peripheral blood of mice was collected retro-orbitally. To analyze the peripheral blood of mice, red blood cells were first lysed and then stained for myeloid (rat anti-Gr-1 APC-Cy7, BD, 557661, rat anti-Mac-1 APC, eBioscience, 17-0112-83) or lymphoid lineages (rat anti-B220 FITC, eBioscience, 11-0452-82, hamster anti-CD3 PE-Cy7, eBioscience, 25-0031-82).
Bone marrow was flushed from femurs and cellularity was quantified with 3% acetic acid in methylene blue (STEMCELL). The lineage cell compartment of the bone marrow was analyzed by staining for myeloid and lymphoid lineages as in the peripheral blood. The HSPC compartment was analyzed by staining for Lineage (biotin-Ter-119, -Mac-1, -Gr-1, -CD4, -CD8α, -CD5, -CD19 and -B220 (eBioscience, 13-5921-85, 13-0051-85, 13-5931-86, 13-0112-86, 13-0452-86, 13-0041-86, 13-0081-86, 13-0193-86) followed by staining with streptavidin-PE-TexasRed (Invitrogen, SA1017), rat anti-cKit APC-Cy7 (eBioscience, 47-1171-82), rat anti-Sca1 PerCP-Cy5.5 (eBioscience, 45-5981-82), hamster anti-CD48 APC (eBioscience, 17-0481-82) and rat anti-CD150 PE-Cy7 (Biolegend, 115914). All flow cytometry analysis was performed on a BD LSR Fortessa flow cytometer and was analyzed on FlowJo v10.5.3 for MAC.

Zhong L., Yao L., Tower R.J., Wei Y., Miao Z., Park J., Shrestha R., Wang L., Yu W., Holdreith N., Huang X., Zhang Y., Tong W., Gong Y., Ahn J., Susztak K., Dyment N., Li M., Long F., Chen C., Seale P, & Qin L. (2020). Single cell transcriptomics identifies a unique adipose lineage cell population that regulates bone marrow environment. eLife, 9, e54695.

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