pbs phosphate buffered saline 10x ph 7 4  (Thermo Fisher)

Journal: The Journal of Infectious Diseases

Article Title: Bacterial Superantigens Expand and Activate, Rather than Delete or Incapacitate, Preexisting Antigen-Specific Memory CD8+ T Cells

doi: 10.1093/infdis/jiy647

Figure Lengend Snippet: In vivo administration of Staphylococcal enterotoxin B (SEB) expands vaccine-elicited, Vβ8.1/8.2 + memory TCD8 in mice without compromising their effector functions. A , BALB/c mice were inoculated intraperitoneally with influenza A virus (IAV) to generate memory TCD8 and, 2 months later, received SEB (50 μg) or phosphate-buffered saline (PBS) intraperitoneally. Three days later, splenic NP 147 -specific TCD8 were enumerated by intracellular cytokine staining. B , Splenocytes from naive BALB/c mice were cultured for 3 days in the presence or absence of SEB, and the frequencies of indicated T cell receptor (TCR) Vβ + cells among the total number of TCD8 were determined. C , The expression of TCR Vβ8.1/8.2 by NP 147 -specific TCD8 was also examined in IAV-primed mice in which PBS or SEB were injected. Open and filled histograms correspond to staining with an anti–mouse TCR Vβ8.1/8.2 monoclonal antibody and isotype control, respectively. D , Cognate and irrelevant target cells were prepared by pulsing naive syngeneic splenocytes with TYQRTRALV (the NP 147 peptide epitope of IAV) and RPQASGVYM (the NP 118 peptide epitope of LCMV), respectively. Target cells were labeled with 2 different doses of carboxyfluorescein succinimidyl ester (CFSE), washed, mixed in equal numbers, and injected intravenously into IAV-primed BALB/c mice that later received either SEB (50 μg intraperitoneally) or PBS. Four hours later, mice were euthanized, and target cell populations were tracked on the basis of their differential CFSE staining intensities in each spleen. Histograms from representative mice are shown. Percent specific cytotoxicity was calculated as described in Methods. Each open and filled circle in panel D represents an individual mouse. Error bars in panels A and D represent standard errors of the mean. IFN-γ, interferon γ. ** P

Article Snippet: Splenocytes were suspended at 2 × 106 cells per mL of complete medium in T75 flasks, to which SEB (500 ng/mL) or phosphate-buffered saline (PBS) was added.

Techniques: In Vivo, Mouse Assay, Staining, Cell Culture, Expressing, Injection, Labeling

Journal: Theranostics

Article Title: Triple-Modal Imaging of Magnetically-Targeted Nanocapsules in Solid Tumours In Vivo

doi: 10.7150/thno.11918

Figure Lengend Snippet: Radio-labelling and in vitro serum stability of NC and m- NC. The NC and m- NC were prepared by single emulsification/solvent evaporation method. PLGA 18KDa -PEG 3.5KDa -DTPA was incorporated in the formulation at 10% (w/w). The radio-labelling reaction was carried out for 30 min in 0.2 M ammonium acetate (pH 5.5) and quenched by the addition of 0.1M EDTA of 1/20 (v/v) of the total volume. a , Radio-labelling efficiency was evaluated immediately after labelling and 100% radio-labelling efficiency was obtained for both types of NCs whereas NCs without incorporation of DTPA showed no radio-labelling. b , Serum stability was tested by incubation of NC- 111 In and m- NC- 111 In in serum or PBS at 37 °C up to 24 h. Both types of NC exhibited high stability in serum and PBS indicated by presence of low amount of 111 In-EDTA.

Article Snippet: Advanced RPMI-1640 media, penicillin-Streptomycin 100x, Trypsin-EDTA (1X) with Phenol red Glutamax™ supplement, phosphate buffered saline PBS (10x, pH 7.4) were obtained from Gibco, Invitrogen (UK).

Techniques: In Vitro, Evaporation, Incubation

Journal: Oncoimmunology

Article Title: Contribution of annexin A1 to anticancer immunosurveillance

doi: 10.1080/2162402X.2019.1647760

Figure Lengend Snippet: Wild type (wt), Annexin A1 ( AnxA1 −/- ), Formyl peptide receptor 1 ( Fpr1 −/- ) and mitochondrial methionyl-tRNA formyltransferase ( Mtfmt −/- ) deficient murine MCA205 fibrosarcoma (a,c) or TC-1 non small cell lung carcinoma (b,d) cells were treated with phosphate-buffered saline (PBS), 2 μM mitoxantrone (MTX), 25 or 20 µM doxorubicin (DOXO) for 18 and 24 h, respectively. Cells were subsequently assessed by cytofluorometric immunodetection of calreticulin (CALR) exposure. Results from one representative experiment out of three independent ones yielding similar results are reported in (b,d). Data are represented as mean ± SD. Representative cytofluorometric profiles for one sample out of three independent ones belonging to at least 3 independent experiments are reported in (a,c). **** p

Article Snippet: In brief, 1 × 105 cells were collected, washed in phosphate-buffered saline (PBS), pelleted and resuspended in 50 µL staining buffer containing FITC-conjugated AnnexinV antibody.

Techniques: Immunodetection

Journal: Oncoimmunology

Article Title: Contribution of annexin A1 to anticancer immunosurveillance

doi: 10.1080/2162402X.2019.1647760

Figure Lengend Snippet: (a,c) Wild type (WT) immunocompetent C57BL/6 mice were inoculated subcutaneously (s.c.) with Annexin A1 deficient ( AnxA1 −/- ) murine MCA205 fibrosarcoma (a) or TC-1 non small cell lung cancer cells (c), respectively. (b,d) Formyl peptide receptor 1 knock out ( Fpr1 −/- ) mice were inoculated s.c . with WT murine MCA205 fibrosarcoma (b) or TC-1 cells (d), respectively. Tumor size was routinely assessed. When tumor became palpable, mice received intratumorally (i.t.) either recombinant calreticulin (rCALR) or doxorubicin (DOXO), alone or in combination or an equivalent volume of phosphate-buffered saline (PBS). From left to right: (1) average (±SEM) tumor growth curves from one representative experiment of two; (2) tumor size distribution at day 25 (MCA205 Anxa1 −/- , (a)), day 24 (MCA205 WT, (b)), day 21 (TC-1 Anxa1 −/- , (c)) or day 18 (TC-1 WT, (d)) of data shown in (1); (3) individual growth curves from mice injected with DOXO alone or combined with rCALR. **** p

Article Snippet: In brief, 1 × 105 cells were collected, washed in phosphate-buffered saline (PBS), pelleted and resuspended in 50 µL staining buffer containing FITC-conjugated AnnexinV antibody.

Techniques: Mouse Assay, Knock-Out, Recombinant, Injection

Journal: PLoS ONE

Article Title: A high-throughput imaging and quantification pipeline for the EVOS imaging platform

doi: 10.1371/journal.pone.0236397

Figure Lengend Snippet: Fluorescence produced by cell media. a) Representative tiled-images of wells containing different media (phenol red-free ‘ClearDMEM;’ standard ‘DMEM’ containing phenol red; ‘DMEM/F12;’ phosphate buffered saline ‘PBS;’ ‘‘M87’ breast epithelial medium; and FlouroBrite ™ ‘Imaging DMEM’ b) Fluorescent intensity within the colored circles (shown in 2a) was measured by ImageJ (n = 6 for each condition). * statistically significant (p

Article Snippet: To circumvent this, a simple solution was to replace the DMEM with phosphate buffered saline (PBS).

Techniques: Fluorescence, Produced, Imaging