Pbs buffer

Manufactured by Corning

Sourced in United States

PBS (Phosphate-Buffered Saline) is a commonly used buffer solution that maintains a stable pH and osmotic environment for various biological applications. It is composed of a combination of sodium phosphate and sodium chloride, which helps to preserve the structure and function of biomolecules and cells. PBS is widely used in laboratories for a variety of purposes, including cell culture, protein purification, and immunoassays.

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Lab products found in correlation

3 isobutyl 1 methylxanthine, by Merck Group (2 mentions) β mercaptoethanol, by Thermo Fisher Scientific (2 mentions) Thioglycolate medium, by BD (2 mentions) Proteinase k, by Thermo Fisher Scientific (2 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Streptomycin, by Thermo Fisher Scientific (1 mentions) Iscove s modified dulbecco s medium, by Thermo Fisher Scientific (1 mentions) L glutamine, by Thermo Fisher Scientific (1 mentions) Ivis lumina xr, by PerkinElmer (1 mentions) Living image software, by PerkinElmer (1 mentions) Progesterone, by Merck Group (1 mentions) 8 bromo camp, by Cayman Chemical (1 mentions) Camp elisa kit, by Cayman Chemical (1 mentions) Horseradish peroxidase conjugated anti mouse and anti rabbit igg, by Cell Signaling Technology (1 mentions) Nahco3, by Merck Group (1 mentions) Bovine serum albumin (bsa), by Merck Group (1 mentions) Mojosort mouse pan b cell isolation kit, by BioLegend (1 mentions) Co2 incubator, by Thermo Fisher Scientific (1 mentions) Human serum albumin (hsa), by Irvine Scientific (1 mentions) Concanavalin a (cona), by Merck Group (1 mentions)

13 protocols using pbs buffer

Cell Culture and Lymphoma Cell Preparation for Northern Blot

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HeLa cells were cultured in DMEM with glutamax (Dulbeccos's modified eagle medium with glutamax, Gibco 10569) supplemented with 10% fetal bovine serum and 1% of 100 units/ml penicillin/streptomycin (100x stock). The OCI-Ly3, OCI-Ly1, and OCI-Ly19 cell lines were obtained from Dr. Minden (University of Toronto) and cultured in Iscove's Modified Dulbecco's Medium (Life Technologies) containing 20% Fetal Bovine Serum (FBS; HyClone), and 100 units/ml penicillin and 100 μg/ml streptomycin (Gibco). The BJAB, Ramos, Raji, SU-DHL-4 cell lines were obtained from the American Type Culture Collection (ATCC) and cultured in RPMI 1640 Medium containing 2 mM L-Glutamine (Gibco), 10% FBS, and 100 units/ml penicillin and 100 μg/ml streptomycin. All human lymphoma cell lines were cultured to approximately 1 × 10⁶ cell/ml at 37°C with 5% CO₂ in T-175 flasks. Cells were removed, centrifuged (7 min, 270 rcf), and resuspended in PBS buffer (Corning Cellgro). Cells were counted, and 1.8 × 10⁷ cells were centrifuged. Following removal of the supernatant, cell pellets were frozen at −80°C until further processing for Northern blot.

Jin H.Y., Gonzalez-Martin A., Miletic A.V., Lai M., Knight S., Sabouri-Ghomi M., Head S.R., Macauley M.S., Rickert R.C, & Xiao C. (2015). Transfection of microRNA Mimics Should Be Used with Caution. Frontiers in Genetics, 6, 340.

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Immobilization and Detection of IgG using Gold Nanoparticles

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IgG was immobilized to an activated sensor surface by incubating the sensor in 100 μL of prepared rabbit IgG solution at room temperature for 10 min, followed by washing it with 250 μL of PBST buffer (0.05% Tween 20 in PBS buffer, Corning Cellgro) three times. Residual activated binding sites were blocked by 250 μL BSA solution (1% w/v) for 5 min and then washed with 250 μL of PBST buffer three times. 100 μL solution of gold nanoparticles coated with goat anti-rabbit IgG was added, and the binding of the gold nanoparticles to IgG was tracked in real time.

Jing W., Wang Y., Yang Y., Wang Y., Ma G., Wang S, & Tao N. (2019). Time-resolved Digital Immunoassay for Rapid and Sensitive Quantitation of Procalcitonin with Plasmonic Imaging. ACS nano, 13(8), 8609-8617.

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Bioluminescent Imaging of Tumor Cells

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The bioluminescent imaging was performed using an IVIS Lumina XR (Caliper Life Science). The images were analyzed by Living Image software (Caliper Life Science). For invitro imaging, the MDA-MB-231-Luc cells were plated in a 6-well plate with indicated density, and then D-luciferin K (Gold Biotechnology) was added to the plate, to the final concentration 150 µg/ml. The cells were incubated 5 min before imaging. The luminescent exposure time is 1 s/plate. For invivo imaging, the mice received I.P. injection of D-luciferin K (150 mg/kg) in PBS buffer (Cellgro). After 10–15 min, mice were anesthetized by ∼2% isoflurane through inhalation and immediately imaged with 1 s the luminescent exposure time. Tumor volumes were represented by fluorescence intensity expressed in photons/s. Background bioluminescence invivo was approximately 9×10⁷ photons/s.

Huang F, & Mazin A.V. (2014). A Small Molecule Inhibitor of Human RAD51 Potentiates Breast Cancer Cell Killing by Therapeutic Agents in Mouse Xenografts. PLoS ONE, 9(6), e100993.

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Cyclic AMP Signaling Regulation Assay

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3-Isobutyl-1-methylxanthine (IBMX), bovine serum albumin (BSA), NaHCO₃ and Progesterone were purchased from Sigma-Aldrich (St. Louis, MO, United States). 8-Bromo-cAMP (sodium salt) and cAMP Elisa kit from Cayman Chemical (Ann Arbor, MI, United States). β-mercaptoethanol from Thermo Fisher Scientific (Waltham, MA, United States). PBS buffer was purchased from Corning (Radnor, PA, United States). Rabbit monoclonal anti-phospho-PKA substrates (anti-pPKA substrates) (clone 100G7E) and Horseradish peroxidase-conjugated anti-mouse and anti-rabbit IgG, were purchased from Cell Signaling Technology (Danvers, MA). Mouse anti-phospho Tyrosine (anti-pTyr) (clone 4G10) was from EMD Millipore.

Ritagliati C., Ayoub S., Balbach M., Buck J, & Levin L.R. (2023). In vivo characterization of sAC null sperm. Frontiers in Cell and Developmental Biology, 11, 1134051.

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Visualizing Intracellular CXCL11 in B Cells

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For visualizing intracellular CXCL11 expressed by B cells, B cells were
isolated from collagenase-digested tumor tissues using the MojoSort Mouse Pan
B-cell Isolation Kit (Cat# 480051, Biolegend). The enriched B cells were fixed
in cold methanol overnight and then in blocking buffer (1X PBS / 5% normal serum
(#13551 Stemcell) / 0.3% Triton™ X-100, CAT#T8787, Sigma) for 60 minutes.
Cells were co-stained with FITC-anti-mouse CXCL11 (aa22–100, Cat#
LS-C693458, LSBio) or its FITC-IgG Isotype Control (Cat# LS-C149376, LSBio) and
PE-anti-mouse B220 (Cat^# 50–0452-U100, Biolegend) at a 1:200
dilution in buffer (1X PBS / 1% BSA / 0.3% Triton™ X-100) and incubated
overnight at 4°C. The stained cells were then washed with PBS buffer
(Cat# 46–013-CM, Corning) and mounted on slides. PE-labeled
B220⁺ cells co-expressing CXCL11 (based upon co-staining for
FITC) were visualized by LSM 510 Meta confocal microscopy and photographed.
Images were analyzed in ImageJ using the corrected total cell fluorescence
(CTCF) method and the provided region of interest (ROI) tool. CTCF of CXCL11 was
calculated for the cytosol by the following formula: CTCF = Integrated Density
− (Area of Region of Interest * Fluorescence of background reading).

Yang J., Yan C., Vilgelm A.E., Chen S.C., Ayers G.D., Johnson C.A, & Richmond A. (2020). Targeted Deletion of CXCR2 in Myeloid Cells Alters the Tumor Immune Environment to Improve Antitumor Immunity. Cancer immunology research, 9(2), 200-213.

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Clonal Cell Population Expansion

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Transfected cells were washed with 500 μL of PBS buffer (Corning, USA) and resuspended at the density of 8 cells/mL with a total volume of 50 mL. 100 μL of the cell suspension was transferred into the wells of the 96 well tissue culture plates to ensure each well contained a single cell. The plates were incubated at 37 °C, 5% CO₂ incubator (Thermo Scientific, USA). The plates were then scanned for single cell colonies as soon as small aggregates of cells are visible under a 4× microscope (usually after first week, depending on the growth rate of the cell line) to ensure the cell colonies were derived from a single cell. The cells were incubated for an additional 2–3 weeks to expand the clonal populations for further analysis and characterization.

Yin Y., Reed E.F, & Zhang Q. (2019). Integrate CRISPR/Cas9 for protein expression of HLA-B*38:68Q via precise gene editing. Scientific Reports, 9, 8067.

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Reagents for Cell Culture Media

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Reagents for media preparation were purchased from Sigma-Aldrich. 2-deoxyglucose (2-DG), 3-Isobutyl-1-methylxanthine (IBMX), antimycin A, BSA, concanavalin A (ConA), dibutyryl-cAMP (db-cAMP), progesterone, lectin from Pisum sativum FITC-conjugated (PSA-FITC) and lectin from Arachis hypogaea FITC-conjugated (PNA-FITC) were purchased from Sigma-Aldrich, rotenone (rot) from Cayman chemicals, HSA from Irvine Scientific and PBS buffer from Corning.

Balbach M., Ghanem L., Violante S., Kyaw A., Romarowski A., Cross J.R., Visconti P.E., Levin L.R, & Buck J. (2023). Capacitation induces changes in metabolic pathways supporting motility of epididymal and ejaculated sperm. Frontiers in Cell and Developmental Biology, 11, 1160154.

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Immunolabeling and Histological Analysis of Lens Cryosections

Cited 1 time

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Lenses are isolated and fixed for 2 or 24 hrs at 4°C in 4% paraformaldehyde, washed in PBS, and then cryoprotected (30% sucrose) prior to freezing in Polyfreeze Tissue Freezing Media Red (Polyscience #25115). 20μm thick serial cryosections were cut using a Microm HM 550 Cryostat, and only the central sections used for these studies. Immunolabeling was performed as described previously [41 (link)]. Briefly, the sections were permeabilized with 0.25% Triton X-100 in PBS buffer (Corning) for 30 min and the incubated in block buffer (5% goat serum, 1% BSA in PBS) for 1 hr prior to incubation in primary antibody for either 3 hrs at 37 C or overnight at 4 C, followed by incubation with secondary antibody for 2 hrs at 37 C (Jackson ImmunoResearch Laboratories). F-actin was localized with Alexa 647-conjugated phalloidin (Invitrogen-Molecular Probes). Nuclei were stained with DAPI (Biolegend). For histological analyses, lenses were fixed in 4% paraformaldehyde, cryoprotected (30% sucrose) for 24 hrs and frozen with tissue freezing media prior to cryosectioning. Sections were stained with hematoxylin and eosin.
Images were acquired with a Nikon Eclipse Ti microscope with a Nikon Photometrics Cool Snap HQ camera.

Gheyas R., Ortega-Alvarez R., Chauss D., Kantorow M, & Menko A.S. (2022). Suppression of PI3K signaling is linked to autophagy activation and the spatiotemporal induction of the lens Organelle Free Zone. Experimental cell research, 412(2), 113043.

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Spermatogenesis Assay and Visualization

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3-Isobutyl-1-methylxanthine (IBMX), BSA, dibutyryl-cAMP (db-cAMP), hyaluronidase, lectin from Pisum sativum FITC-conjugated (PSA-FITC) and lectin from Arachis hypogaea FITC-conjugated (PNA-FITC) were purchased from Sigma-Aldrich, ionomycin from Tocris, β-mercaptoethanol from Gibco, and hormones from ProSpec. PBS buffer was purchased from Corning, DMEM, and 0.5 M EDTA, pH 8.0 from Thermo Fisher Scientific, FBS from Avantor Seradigm, and polyethylene glycol 400 (PEG 400) from Merck Millipore.
sAC-overexpressing rat 4-4 cells were generated and functionally authenticated in our laboratory as previously described^{33 (link)} and grown in DMEM + 10% FBS. Cells were maintained at 37 °C in 5% CO₂ and were periodically checked for mycoplasma contamination.
8–10 week-old C57BL/6J male and female mice were purchased from Jackson Laboratories and allowed to acclimatize before use. All animals were maintained at a 12 hr light/dark cycle, temperatures of 64–79 °F (~18–26 °C) with 40–60% humidity in accordance with the NIH Guide for the Care and Use of Laboratory Animals. Animal experiments were approved by Weill Cornell Medicine’s Institutional Animal Care and Use Committee (IACUC).

Balbach M., Rossetti T., Ferreira J., Ghanem L., Ritagliati C., Myers R.W., Huggins D.J., Steegborn C., Miranda I.C., Meinke P.T., Buck J, & Levin L.R. (2023). On-demand male contraception via acute inhibition of soluble adenylyl cyclase. Nature Communications, 14, 637.

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Optimizing Cellular Assays with Diverse Reagents

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RPMI 1640 medium was purchased from Lonza (Basel, Switzerland). Media supplements, penicillin, streptomycin were obtained from Sigma (MilliporeSigma, St. Louis, MO, USA). Everolimus, 10 mM solution in DMSO, was purchased from Selleck Chemicals (Houston, TX, USA). Bovine hemin powder (purity ≥ 90%) was purchased from Sigma (MilliporeSigma, St. Louis, MO, USA), and further dissolved in a 1.4 M NaOH aqueous solution to obtain 7.7 mM hemin stock solutions. Doxorubicin and cisplatin were made available for research courtesy of EBEWE Pharma (EBEWE Pharma GmbH Nfg. KG, Unterach am Attersee, Austria). Phycoerythrin-conjugated mouse anti-human fetal hemoglobin (clon 2D12), mouse IgG1, kappa isotype control, BD Cytofix/Cytoperm™ Fixation/Permeablization Kit were obtained from Becton Dickinson. PBS buffer were purchased from Corning (New York, NY, USA).

Jakubowska J., Pawlik B., Wyka K., Stolarska M., Kotulska K., Jóźwiak S., Młynarski W, & Trelińska J. (2021). New Insights into Red Blood Cell Microcytosis upon mTOR Inhibitor Administration. International Journal of Molecular Sciences, 22(13), 6802.

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