Acsa 2 apc

Manufactured by Miltenyi Biotec

The ACSA-2-APC is a fluorescently-labeled antibody produced by Miltenyi Biotec. It is designed for the detection and analysis of cellular antigens using flow cytometry.

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Lab products found in correlation

Cytoflex cytometer, by Beckman Coulter (1 mentions) Cd11b fitc, by Miltenyi Biotec (1 mentions) Ly6g apc, by BD (1 mentions) Gentlemacs dissociator, by Miltenyi Biotec (1 mentions) Trizol reagent, by Qiagen (1 mentions) Propidium iodide, by Merck Group (1 mentions) Cytexpert software 2, by Beckman Coulter (1 mentions) Mouse brain slicer matrix, by Zivic Instruments (1 mentions) Facsaria 3, by BD (1 mentions) Adult brain dissociation kit, by Miltenyi Biotec (1 mentions) Cd11b bv421, by BD (1 mentions) Fixable viability stain bv510, by BD (1 mentions) Fc blocked, by BioLegend (1 mentions) Rlt plus, by Qiagen (1 mentions)

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ACSA-2 (8 citations)

2 protocols using acsa 2 apc

Isolation and Analysis of Brain Immune Cells

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On day 1 after ICH, mice were anesthetized with 3% isoflurane and were perfused with cold phosphate-buffered saline. Brains were cut into 1-mm-thick coronal sections with a mouse brain slicer matrix (Zivic Instruments, Pittsburgh, PA). Four 1-mm-thick brain sections of the left striatum with hemotoma were collected. The striatum was dissociated with GentleMACS Dissociator (Miltenyi Biotec, Auburn, CA) as described previously^{28 (link),30 (link)}. The final cell suspension were incubated with the primary antibodies: CD11b-FITC (Miltenyi Biotec, 130–113-234) and ACSA-2-APC (Miltenyi Biotec, 130–117-535) or and Ly6G-APC (Pharmingen, 560,599) for 30 min at 4ºC. The corresponding isotype antibodies were used as negative control. Propidium iodide (Sigma, St. Louis, MO) staining was used to exclude dead cells. Cell supernatants were analyzed by CytoFLEX cytometer (Beckman Coulter, Indianapolis, IN) with CytExpert software 2.0 (Beckman Coulter). Macrophage/microglia cells were distinguished as CD11b-FITC⁺ / ACSA-2-APC^- population. Astrocytes were distinguished as CD11b-FITC^- / ACSA-2-APC⁺ population. Neutrophils were distinguished as CD11b-FITC⁺ / Ly6G-APC⁺ population. Gated cells were collected into TRIzol reagent (Qiagen, 217,004) for mRNA extraction and real-time PCR.

Han X., Ren H., Nandi A., Fan X, & Koehler R.C. (2021). Analysis of glucose metabolism by 18F-FDG-PET imaging and glucose transporter expression in a mouse model of intracerebral hemorrhage. Scientific Reports, 11, 10885.

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Isolation and Purification of CNS Cell Types

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CNS cell types were isolated by cell sorting using previously established protocols (Nugent et al., 2020 (link); Ullman et al., 2020 ). Briefly, to prepare a single cell suspension, brain tissue was dissected and processed into a single cell suspension according to the manufacturers’ protocol using the adult brain dissociation kit (Miltenyi Biotec 130–107-677). Cells were Fc blocked (Biolegend #101320, 1:100) and stained for flow cytometric analysis with Fixable Viability Stain BV510 (BD Biosciences #564406, 1:100) to exclude dead cells, CD11b-BV421 (BD Biosciences 562605, 1:100), ACSA2-APC (Miltenyi #130–117-386, 1:100), and Thy1-PE (R&D #FAB7335P, 1:100). Cells were washed with PBS/1% BSA and strained through a 100μm filter before sorting CD11b+ microglia, ACSA2+ astrocytes, and Thy1+ neurons on a FACS Aria III (BD Biosciences) with a 100μm nozzle. Sorted cells were collected directly into MS grade methanol with added internal standards for lipidomic and metabolomic analysis or Qiagen RLT-plus + β-mercaptoethanol for RNA analysis.

Logan T., Simon M.J., Rana A., Cherf G.M., Srivastava A., Davis S.S., Yoon Low R.L., Chiu C.L., Fang M., Huang F., Bhalla A., Llapashtica C., Prorok R., Pizzo M.E., Calvert M.E., Sun E.W., Hsiao-Nakamoto J., Rajendra Y., Lexa K.W., Srivastava D.B., van Lengerich B., Wang J., Robles-Colmenares Y., Kim D.J., Duque J., Lenser M., Earr T.K., Nguyen H., Chau R., Tsogtbaatar B., Ravi R., Skuja L.L., Solanoy H., Rosen H.J., Boeve B.F., Boxer A.L., Heuer H.W., Dennis M.S., Kariolis M.S., Monroe K.M., Przybyla L., Sanchez P.E., Meisner R., Diaz D., Henne K.R., Watts R.J., Henry A.G., Gunasekaran K., Astarita G., Suh J.H., Lewcock J.W., DeVos S.L, & Di Paolo G. (2021). Rescue of a lysosomal storage disorder caused by Grn loss-of-function with a brain penetrant progranulin biologic. Cell, 184(18), 4651-4668.e25.

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