Cells were stained with fluorescein-conjugated mAbs to mouse CD3α (145-2C11), CD4 (RM4–5), CD8α (53–6.7), CD11c (HL3), CD40 (3/23), CD44 (IM7), CD45.1 (A20), CD62L (MEL-14), CD80 (16-10A1), CD86 (GL1), CD154 (MR1), I-A/I-E (M5/114.15.2), B220 (RA3-6B2), Vα2 TCR (B20.1), γδTCR (GL3), IFN-γ (XMG1.2), IL-17A (TC11-18H10), isotype-matched control mAb (BD Biosciences), CD209b (22D1), CD253/TRAIL (N2B2) (eBioscience), CD11b (M1/70), CD178/FasL (MFL3), F4/80 (BM8), Ly6C (HK1.4), Ly6G (1A8), γδTCR (GL3) (Biolegend), mPDCA-1/BST2 (JF05-1C2.4.1) (Miltenyi Biotec), H-2Kb OVA tetramer (MBL) and Siglec-H (440c; Hycult Biotechnology). For the intracellular expression of cytokines, cells were incubated for 4 hrs with phorbol 12-myristate 13-acetate (PMA, 50 ng/ml; Sigma-Aldrich) plus ionomycin (500 ng/ml; Sigma-Aldrich) or OVA257–264 peptide (SIINFEKL; 10 μM) plus GolgiPlug (BD Biosciences) during the final 2 hrs. Subsequently, the cells were resuspended in Fixation-Permeabilization solution (BD Cytofix/Cytoperm kit; BD Biosciences) and intracellular cytokine staining was carried out according to the manufacturer’s directions. Fluorescence staining was analyzed with a FACSCalibur flow cytometer and CELLQuest software (both from BD Biosciences).
Mpdca 1 bst2
Manufactured by Miltenyi Biotec
The MPDCA-1/BST2 is a lab equipment product from Miltenyi Biotec. It is used to detect and enrich for CD317 (BST2, tetherin) positive cells.
Automatically generated - may contain errors
Lab products found in correlation
Tcr vα2, by BD (1 mentions)
γδtcr gl3, by BD (1 mentions)
Isotype matched control mab, by BD (1 mentions)
Ova257 264 peptide, by BD (1 mentions)
Ionomycin, by Merck Group (1 mentions)
Phorbol 12 myristate 13 acetate pma, by Merck Group (1 mentions)
Golgiplug, by BD (1 mentions)
Cytofix cytoperm kit, by BD (1 mentions)
Ifn γ, by BD (1 mentions)
Anti cd11c, by Thermo Fisher Scientific (1 mentions)
Lsrfortessa, by BD (1 mentions)
Facsaria, by BD (1 mentions)
Sirpα, by BD (1 mentions)
2 protocols using mpdca 1 bst2
1
Flow Cytometric Phenotyping and Cytokine Analysis
2
Identification and Sorting of DC Subsets
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cells were stained with the following fluorochrome- or biotin-conjugated antibodies obtained from eBioscience (or another manufacturer, as indicated): anti-CD11c (N418), MHC II (M5/114.15.2), CD11b (M1/70), CD8α (53–6.7), Gr-1 (RB6-8C5), TCRβ (H57-597), CD3 (17A2), B220 (RA3-6B2), NK1.1 (PK136), Ter119 (TER-119), CD49b (DX5), CD4 (RM4-5 and GK1.5), FoxP3 (FJK-16s), CD24 (M1/69), Flt3 (A2F10), c-Kit (2B8), Sca-1 (D7), CD115 (AFS98), IL7-Rα (A7R34), CD45.1 (A20), CD45.2 (104), PD-L1 (MIH5), CD86 (GL1), DEC205 (NLDC-145; BioLegend), mPDCA-1/Bst2 (Miltenyi Biotec), Sirpα (P84, BD), CD80 (16-10A1), CD40 (1C10), and MHC I (AF6-88.5.5.3). Intracellular staining of FoxP3 was performed per the manufacturer’s instructions (eBioscience). Cell acquisition was done on LSR II or LSR Fortessa (BD) and analysis was done using FlowJo (Tree Star).
For sorting, spleens were harvested from Flt3+/+ or Flt3ITD/+ FVB-B6 F1 mice and sorted on either FACSAria (BD) or MoFlo (Beckman Coulter) instruments. DC populations were sorted as follows: pDC (Bst2+ B220+), cDCs (CD11chi MHCII+ B220−) and subsets thereof; canonical CD8+ (CD8+ SSChi DEC205+ or CD86+); noncanonical CD8+ (CD8+ SSClo DEC205− or CD86−); Esamhi (CD11b+ Esamhi); and Esamlo (CD11b+ Esamlo).
For sorting, spleens were harvested from Flt3+/+ or Flt3ITD/+ FVB-B6 F1 mice and sorted on either FACSAria (BD) or MoFlo (Beckman Coulter) instruments. DC populations were sorted as follows: pDC (Bst2+ B220+), cDCs (CD11chi MHCII+ B220−) and subsets thereof; canonical CD8+ (CD8+ SSChi DEC205+ or CD86+); noncanonical CD8+ (CD8+ SSClo DEC205− or CD86−); Esamhi (CD11b+ Esamhi); and Esamlo (CD11b+ Esamlo).
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!