Cd3ε pe

Manufactured by BioLegend

CD3ε-PE is a fluorescently labeled antibody that binds to the CD3ε subunit of the T cell receptor complex. It is commonly used in flow cytometry applications to identify and enumerate T cells within a sample.

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PE anti-mouse CD3ε (5 citations) Anti-mouse CD3ε-PE/Cy7 (5 citations) CD3ε PE-Cy7, (3 citations) PE antimouse CD3ε antibody (2 citations) PE-CD3ε (145-2C11; (2 citations) CD3ε (PE/Cy7; clone 145.2C11; (2 citations)

4 protocols using cd3ε pe

Multicolor Flow Cytometry of Brain Myeloid Cells

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BSLs purified from perfused brain tissue of moribund C57BL/6 mice on day 6 post-infection were stained with TCRβ-FITC, CD3ε-PE, CD4-PerCP, CD4-PerCP/Cy5.5, CD8-APC/Cy7, CD11b-APC, CD14-PE/Cy7, and F4/80-Brilliant Violet 421™ antibodies purchased from Biolegend (San Diego, CA) and fixable viability dye eFluor^®506 (eBioscience, San Diego, CA). An Amnis^® ImageStream^®X MKII (Amnis Merck-Millipore, Seattle, WA) equipped with five lasers (355, 405, 488, 561, and 642nm) was used for acquisition. A minimum of 30,000 events were collected at 40X magnification using the INSPIRE^® software (Amnis Merck-Millipore, Seattle, WA). Analysis was performed using IDEAS® 6.1 software (Amnis Merck-Millipore, Seattle, WA). Focused events were first gated using the Gradient RMS feature. Singlets were then gated by aggregate discrimination using aspect ratio and area. Analysis was then performed on gated live CD11b^highCD14⁺F480⁺TCRβ⁺CD3ε−CD4−CD8− cells.

Oakley M.S., Chorazeczewski J.K., Aleshnick M., Anantharaman V., Majam V., Chawla B., Myers T.G., Su Q., Okoth W.A., Takeda K., Akue A., KuKuruga M., Aravind L, & Kumar S. (2018). TCRβ-expressing macrophages induced by a pathogenic murine malaria correlate with parasite burden and enhanced phagocytic activity. PLoS ONE, 13(7), e0201043.

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Boronic Acid-Based Glucose-Responsive Hydrogels

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N-isopropylmethacrylamide (NIPMAAm), 4-(2-acrylamidoethylcarbamoyl)-3-fluorophenylboronic acid (AmECFPBA), N,N′-methylenebisacrylamide (MBAAm), 2,2′-azobisisobutyronitrile (AIBN), FITC-labeled bovine insulin, and all organic solvents (acetone, dichloromethane, ethanol, hexane, toluene, and DMSO) were all purchased from Wako Pure Chemical Industries. NIPMAAm was recrystallized in hexane and then dried in vacuo overnight before use. Antibodies CD45-PE/Cy7, CD11b-FITC, B220-APC, CD3ε-PE, and Ly6G-APC/Cy7 for flow cytometry were purchased from BioLegend. All other reagents were purchased from Sigma-Aldrich or Nacalai Tesque and used as received, unless otherwise noted.

Matsumoto A., Tanaka M., Matsumoto H., Ochi K., Moro-oka Y., Kuwata H., Yamada H., Shirakawa I., Miyazawa T., Ishii H., Kataoka K., Ogawa Y., Miyahara Y, & Suganami T. (2017). Synthetic “smart gel” provides glucose-responsive insulin delivery in diabetic mice. Science Advances, 3(11), eaaq0723.

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Comprehensive Immune Cell Phenotyping

Cited 2 times

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Spleens and adipose tissue (pooled gonadal/renal fat pads) were harvested and processed as described (5 (link), 6 (link)). Cells were stained with antibodies, then results were acquired using a BD LSR II (BD Biosciences) and analyzed with FlowJo software (Tree Star). CD19-BV510 and Vβ8.3-PE (BD Biosciences). I-A^d-A488, CD8α-A700, CD4-APC, CD11c-APC/Cy7, CD3ε-PE, CD11b-PE/Cy7, CD54-FITC, CD40-PE, H²K^d-FITC, I-A^d-PE, CD11c-biotin, SA-APC/Cy7 and TruStain FcX (BioLegend). CD86-APC (eBioscience).

Boi S.K., Buchta C.M., Pearson N.A., Francis M.B., Meyerholz D.K., Grobe J.L, & Norian L.A. (2016). Obesity alters immune and metabolic profiles: new insight from obese-resistant mice on high fat diet. Obesity (Silver Spring, Md.), 24(10), 2140-2149.

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RNA-seq Protocol for Tumor-Infiltrating CD8+ T Cells

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For RNA-sequencing experiments, CD8⁺ TILs and
GFP⁺ tumor cells were sorted from dissociated MC38-PIG
tumors, as were CD8⁺ T cells from dLNs, after staining with the
following antibodies for mouse antigens: CD45.2-AF647 (BioLegend, Clone
#104), CD3ε-PE (BioLegend, Clone#145-2C11), CD8α-BC510
(BioLegend, Clone #53-6.7), CD4-PacBlue (BioLegend, Clone #RM4-5), and
CD11b-PE/Cy7 (BioLegend Clone #M1/70). 20-100K CD8⁺ T cells were
sorted by gating away from debris and then gating for singlets that were
CD45⁺, CD3⁺/CD11b⁻,
CD8⁺/CD4⁻. 200K GFP⁺ tumor cells
were sorted from each sample by gating away from debris and then gating for
singlets that were CD45⁻/GFP⁺. Samples were
sorted into tubes containing R10 supplemented with an extra 10% FBS, stored
on ice, and then spun at 750 g for 10 minutes at
4°C. Cell pellets were resuspended in 30 μL RLT buffer
(Qiagen) with 1% (v/v) 2-mercaptoethanol added and frozen at
−80°C. RNA-seq library preparations were performed as
previously described (Sage et al.,
2016).

Ringel A.E., Drijvers J.M., Baker G.J., Catozzi A., García-Cañaveras J.C., Gassaway B.M., Miller B.C., Juneja V.R., Nguyen T.H., Joshi S., Yao C.H., Yoon H., Sage P.T., LaFleur M.W., Trombley J.D., Jacobson C.A., Maliga Z., Gygi S.P., Sorger P.K., Rabinowitz J.D., Sharpe A.H, & Haigis M.C. (2020). Obesity shapes metabolism in the tumor microenvironment to suppress anti-tumor immunity. Cell, 183(7), 1848-1866.e26.

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