Phosflow perm buffer 3

Manufactured by Thermo Fisher Scientific

Sourced in United States

PhosFlow Perm Buffer III is a buffer solution used for permeabilization and fixation of cells prior to flow cytometric analysis of intracellular phosphoproteins. The buffer aids in preserving the phosphorylation state of proteins within the cells.

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Lab products found in correlation

Anti phospho stat5 py694, by BD (1 mentions) Antiphospho akts473 d9e, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

Perm Buffer (40 citations) Perm Buffer III (2 citations)

2 protocols using phosflow perm buffer 3

Flow Cytometry Staining Protocol

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Staining was performed in the presence of purified Ab to CD16/32 at saturation to block nonspecific staining. For flow analysis, cells were lysed and fixed with 1× PhosphoFlow Lyse/Fix Buffer for 15 min at 37 °C, and then permeabilized with PhosFlow Perm Buffer III on ice for 1 h (all reagents were from eBioscience USA).

Lu Y., Meng R., Wang X., Xu Y., Tang Y., Wu J., Xue Q., Yu S., Duan M., Shan D., Wang Q., Wang H., Billiar T.R., Xiao X., Chen F, & Lu B. (2019). Caspase-11 signaling enhances graft-versus-host disease. Nature Communications, 10, 4044.

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Characterizing Murine Immune Cell Subsets

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Monoclonal antibodies against mouse CD3 (17A2), NKp46 (29A1.4), NK1.1 (PK136), CD127 (SB/199), Ly49A (A1), Ly49H (3D10), Ly49G2 (4D11), Ly49D (4E5), NKG2A (20D5), NKG2D (CX5), KLRG1 (2F1), CD11b (M1/70), CD27 (LG.7F9), Bcl-2 (10C4), E4BP4 (S2M-E19), Eomes (Dan11mag), T-bet (4B10), Ki67 (B56) and isotype controls were purchased from eBioscience. Anti-Ly49C/I (YLI-90) and anti-phospho-STAT5 (pY694) were purchased from BD Biosciences. Anti-phospho-S6 (D57.2.2E). Anti-phospho-AKT T₃₀₈ (D25E6) and anti-phospho-AKT S₄₇₃ (D9E) were obtained from Cell Signaling Technology. For the detection of phosphorylated signaling proteins, NK cell was fixed with Phosflow Lyse/Fix buffer, permeabilized with Phosflow Perm buffer III (eBioscience), and then stained with antibodies. Data were acquired on an LSRII (Becton Dickinson) flow cytometer and analyzed using FlowJo software (Treestar). The expression level was presented as percentage or net MFI, which was determined by subtracting the mean fluorescence intensity of isotype control.

He J., Zhao J., Quan Y., Hou X., Yang M, & Dong Z. (2021). Full Activation of Kinase Protein Kinase B by Phosphoinositide-Dependent Protein Kinase-1 and Mammalian Target of Rapamycin Complex 2 Is Required for Early Natural Killer Cell Development and Survival. Frontiers in Immunology, 11, 617404.

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