Facsaria 2

Manufactured by Takara Bio

The FACSAria II is a flow cytometry instrument designed for high-performance cell sorting. It features advanced optics, digital electronics, and fluidics systems to enable precise and efficient cell analysis and sorting.

Automatically generated - may contain errors

Lab products found in correlation

Lysis buffer, by Takara Bio (3 mentions) Rnase inhibitor, by Takara Bio (3 mentions) Zombie aqua, by BioLegend (3 mentions) Smart seq v4 capture buffer, by Takara Bio (1 mentions) Rnaiso reagent, by Takara Bio (1 mentions) Cd38 pe cy7, by BioLegend (1 mentions) Cd3 apc cy7, by BioLegend (1 mentions) Addavax, by InvivoGen (1 mentions) C57bl 6j mice, by Jackson ImmunoResearch (1 mentions) Igd percp cy5, by BioLegend (1 mentions) Cd71 fitc cy1g4, by BioLegend (1 mentions) Cd3 alexa700, by BioLegend (1 mentions) Cd138 bv421, by Thermo Fisher Scientific (1 mentions) Gl7 percp cy5, by BioLegend (1 mentions) Igd fitc, by BioLegend (1 mentions)

5 protocols using facsaria 2

Isolation and Sequencing of SARS-CoV-2 Specific B Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Peripheral blood samples from convalescent COVID-19 subjects were obtained with informed consent under the approval of the University of Washington Institutional Review Board (Gale Lab; 00009810) and isolated as previously described (Rodda et al., 2021 (link)). Briefly, peripheral blood mononuclear cells were thawed, washed, and stained with a decoy tetramer and then RBD tetramer; bound cells were magnetically enriched (Miltenyi Biotec) and stained with surface antibodies. Single tetramer-positive B cells were index-sorted using a FACS Aria II and collected in 96-well plates containing SMART-Seq v4 capture buffer (Takara Bio) for BCR sequencing.

Hale M., Netland J., Chen Y., Thouvenel C.D., Smith K.N., Rich L.M., Vanderwall E.R., Miranda M.C., Eggenberger J., Hao L., Watson M.J., Mundorff C.C., Rodda L.B., King N.P., Guttman M., Gale M J.r., Abraham J., Debley J.S., Pepper M, & Rawlings D.J. (2022). IgM antibodies derived from memory B cells are potent cross-variant neutralizers of SARS-CoV-2. The Journal of Experimental Medicine, 219(9), e20220849.

+ Open protocol

+ Expand

Alkbh5 Knockout Lin- Cell Sorting

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lin⁻ cells from Alkbh5^fl/fl and Alkbh5^fl/flMx1-Cre mice were sorted using FACS AriaII flow cytometer and resuspended in Trizol (RNAiso Reagent; TaKaRa). All samples were processed and sequenced at Lianchuan Biotechnology Corporation, Hangzhou, China.

Yang B., Liu Y., Xiao F., Liu Z., Chen Z., Li Z., Zhou C., Kuang M., Shu Y., Liu S, & Zou L. (2023). Alkbh5 plays indispensable roles in maintaining self-renewal of hematopoietic stem cells. Open Medicine, 18(1), 20230766.

+ Open protocol

+ Expand

Single-cell Plasmablast Isolation Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

All procedures involving animals were performed in accordance with guidelines of Institutional Animal Care and Use Committee of Washington University in Saint Louis (protocol number 21-0053). Two female C57BL/6J mice (Jackson Laboratories) were immunized intramuscularly and boosted nine weeks later with 15 µg EtpA emulsified in PBS and AddaVax (InvivoGen). Draining iliac and inguinal lymph nodes were collected four days after boosting and single cell suspensions were prepared for plasmablast sorting. Cells were stained for 30 min on ice with CD138-BV421 (281-2, 1:200), IgD-FITC (11-26c.2a, 1:100), CD19-PE (1D3, 1:200), CD38-PE-Cy7 (90, 1:200), Fas-APC (SA367H8, 1:400), CD3-APC-Cy7 (17A2, 1:100), and Zombie Aqua (Biolegend) diluted in PBS supplemented with 2% FBS and 2mM EDTA. Cells were washed twice and single PBs (CD138⁺ CD38^lo CD19^+/lo CD3⁻ live singlet lymphocytes) were sorted using a FACSAria II into 96-well plates containing 2 µL Lysis Buffer (Clontech) supplemented with 1 U/µL rNase inhibitor (NEB) and immediately frozen on dry ice.

Berndsen Z.T., Akhtar M., Thapa M., Vickers T., Schmitz A., Torres J.L., Baboo S., Kumar P., Khatoom N., Sheikh A., Hamrick M., Diedrich J.K., Martinez-Bartolome S., Garrett P.T., Yates JR I.I.I., Turner J.S., Laird R.M., Poly F., Porter C.K., Copps J., Ellebedy A.H., Ward A.B, & Fleckenstein J.M. (2024). Repeat modules and N-linked glycans define structure and antigenicity of a critical enterotoxigenic E. coli adhesin. bioRxiv.

+ Open protocol

+ Expand

Sorting and Profiling Plasmablasts

Check if the same lab product or an alternative is used in the 5 most similar protocols

Staining for sorting was performed using cryo-preserved PBMCs in 2% FBS and 2 mM ethylenediaminetetraacetic acid (EDTA) in PBS (P2). Cells were stained for 30 min on ice with CD20-Pacific Blue (2H7, 1:400), Zombie Aqua, CD71-FITC (CY1G4, 1:200), IgD-PerCP-Cy5.5 (IA6–2, 1:200), CD19-PE (HIB19, 1:200), CD38-PE-Cy7 (HIT2, 1:200), and CD3-Alexa 700 (HIT3a, 1:200), all BioLegend. Cells were washed twice, and single plasmablasts (live singlet CD19⁺ CD3⁻ IgD^lo CD38⁺ CD20⁻ CD71⁺) were sorted using a FACSAria II into 96-well plates containing 2 μL Lysis Buffer (Clontech) supplemented with 1 U/μL RNase inhibitor (NEB) and immediately frozen on dry ice, or bulk sorted into PBS supplemented with 0.05% BSA and processed for single cell RNAseq.

Amanat F., Thapa M., Lei T., Sayed Ahmed S.M., Adelsberg D.C., Carreno J.M., Strohmeier S., Schmitz A.J., Zafar S., Zhou J.Q., Rijnink W., Alshammary H., Borcherding N., Reiche A.G., Srivastava K., Sordillo E.M., van Bakel H., Turner J.S., Bajic G., Simon V., Ellebedy A.H, & Krammer F. (2021). The plasmablast response to SARS-CoV-2 mRNA vaccination is dominated by non-neutralizing antibodies and targets both the NTD and the RBD. medRxiv.

+ Open protocol

+ Expand

Sorting SARS-CoV-2 RBD-Specific Plasma Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Staining for sorting was performed using fresh lymph node single cell
suspensions in PBS supplemented with 2% FBS and 1mM EDTA (P2). Cells were
stained for 30 min on ice with biotinylated recombinant SARS-CoV-2 RBD diluted
in P2, washed twice, then stained for 30 min at 4°C with Fas-PE (Jo2, BD
Pharmingen), CD4-eFluor 780 (GK1.5, eBioscience), CD138-BV421 (281–2),
IgD-FITC (11–26c.2a), GL7-PerCP-Cy5.5, CD38-PE-Cy7 (90), CD19-APC (1D3),
and Zombie Aqua (all Biolegend) diluted in P2. Cells were washed twice and
single SARS-CoV-2 RBD-specific PBs (live singlet CD19⁺CD4⁻ IgD^lo Fas⁺ CD38^loCD138⁺ RBD⁺) and total PBs (live singlet
CD19⁺ CD4⁻ IgD^lo Fas⁺CD38^lo CD138⁺) were sorted using a FACSAria II into
96-well plates containing 2 μL Lysis Buffer (Clontech) supplemented with
1 U/μL RNase inhibitor (NEB) and immediately frozen on dry ice or bulk
sorted into PBS supplemented with 0.05% BSA and processed for single cell
RNAseq.

Alsoussi W.B., Turner J.S., Case J.B., Zhao H., Schmitz A.J., Zhou J.Q., Lei T., Rizk A.A., McIntire K.M., Chen R.E., Hassan A.O., Fox J.M., Winkler E.S., Thackray L.B., Kafai N.M., Amanat F., Krammer F., Watson C.T., Kleinstein S.H., Fremont D.H., Diamond M.S, & Ellebedy A.H. (2020). A potently neutralizing antibody protects mice against SARS-CoV-2 infection. Journal of immunology (Baltimore, Md. : 1950), 205(4), 915-922.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!