P36931

Manufactured by Thermo Fisher Scientific

Sourced in United States, Germany, Japan

The P36931 is a laboratory equipment product from Thermo Fisher Scientific. It is a general-purpose device designed for use in various laboratory settings. The core function of the P36931 is to provide a controlled environment for experiments or sample processing. No further details about the intended use or specific features of this product can be provided while maintaining an unbiased and factual approach.

Automatically generated - may contain errors

Lab products found in correlation

Lsm 780 confocal microscope, by Zeiss (9 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (6 mentions) Lsm 710, by Zeiss (5 mentions) Alexa fluor 488, by Thermo Fisher Scientific (4 mentions) Lsm 510, by Zeiss (4 mentions) A0564, by Agilent Technologies (3 mentions) G2654, by Merck Group (3 mentions) Tb0198, by Sangon (3 mentions) Sc 281692, by Santa Cruz Biotechnology (2 mentions) Ab96920, by Abcam (2 mentions) Dylight 550, by Abcam (2 mentions) Chamber slide, by Thermo Fisher Scientific (2 mentions) Bodipy, by Thermo Fisher Scientific (2 mentions) Lsm 880, by Zeiss (2 mentions) Axioscope 63, by Zeiss (2 mentions) Fluorescence microscope, by Olympus (2 mentions) Alexa fluor 546, by Thermo Fisher Scientific (2 mentions) Ab55051, by Abcam (1 mentions) Dm2500 light microscope, by Leica camera (1 mentions) Prolong gold antifade mounting medium, by Thermo Fisher Scientific (1 mentions)

59 protocols using p36931

Visualizing CD4+ Tissue-Resident Memory T Cells

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To detect CD69, CD103, and RELA/p65 in CD4⁺ T_RM cells, CD4⁺ T_RM cells were fixed with 4% paraformaldehyde for 15 minutes and permeabilized with 0.1% Triton X-100. After blocking with Immunol Staining Blocking Buffer (#P0102, Beyotime), cells were incubated with anti-CD69 (1:100, PA5-102562, Thermo Fisher), anti-CD103 (1:100, MCA708, Bio-Rad), or anti-p65 (1:100, #8242, CST) at 4°C overnight. The next day, cells were probed with Alexa Fluor 594 conjugated donkey anti-mouse immunoglobulin G (#R37115, Invitrogen) or Alexa Fluor 488 conjugated donkey anti-rabbit immunoglobulin G (#R37118, Invitrogen) and then mounted by DAPI (#P36931, Invitrogen). For the staining of lipid droplets, CD4⁺ T_RM cells were incubated with 2 μmol/L BODIPY (#GC42959, Biosharp) for 30 minutes at 37°C. After washing with phosphate-buffered saline 3 times, cells were fixed with 4% paraformaldehyde and mounted with DAPI (#P36931, Invitrogen). The laser scanning confocal microscope (TCS SP8 X, Leica, Germany) was used to capture the images.

Liang G., Huang J., Chen J., Wen X., Li R., Xie H., Zhang Z., Chen Z., Chen Y., Xian Z., He X., Ke J., Lian L., Lan P., Wu X, & Hu T. (2024). Fatty Acid Oxidation Promotes Apoptotic Resistance and Proinflammatory Phenotype of CD4+ Tissue-resident Memory T cells in Crohn’s Disease. Cellular and Molecular Gastroenterology and Hepatology, 17(6), 939-964.

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Quantifying Subcellular Protein Localization

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24 hours post transfection, the coverslip containing transfected cells was washed once with cold 1xPBS, the cells were then fixed with acetone for three minutes and washed with cold 1XPBS for four times. The coverslips were mounted to a clear glass slide using mounting media with DAPI staining (P36931, Thermo Scientific). For each transfection, at least three randomly chosen microscopic fields were imaged. At least 20 cells were qualitatively measured for the subcellular localization as follows: N > C represents predominantly nuclear; N = C represents similar presence in the nucleus and cytoplasm; N < C represents predominantly cytoplasmic. These quantifications are summarized in Table 1.

Yang Y, & Carstens R.P. (2017). Alternative splicing regulates distinct subcellular localization of Epithelial splicing regulatory protein 1 (Esrp1) isoforms. Scientific Reports, 7, 3848.

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Immunohistochemical Analysis of GABA Receptors

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Paraffin section (5 μm) of mouse uterus tissues obtained on days 1–8 were deparaffinized, rehydrated, and then incubated in 3% peroxide in methanol for 15 min at room temperature in order to block endogenous peroxidase activity. After washing three times with PBS, the sections were blocked by 5% albumin solution in PBS at room temperature for 1 h and incubated with GABA_B receptor 1 antibody (1: 300, ab55051, Abcam, UK) at 4 °C overnight. Primary antibody was replaced by 0.5% albumin solution in PBS in control group. After rinsing with PBS three times again, the sections were incubated with the corresponding secondary antibody for another 30 min at room temperature. For Immunofluorescence microscopy, sections were mounted after nuclear counterstain using 4′, 6-diamidino-2-phenylindole (DAPI) (1:1000, no. P36931, Thermo Fisher Scientific), and fluorescence can detect by laser line. The peroxidase on each section were visualized through administration of 3,30-diaminobenzidine solution, and images were obtained under a Leica DM2500 light microscope.

Chen W., Zhang Q., Wang H., Tan D, & Tan Y. (2019). Unique and independent role of the GABAB1 subunit in embryo implantation and uterine decidualization in mice. Genes & Diseases, 8(1), 79-86.

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Fluorescent In Situ Hybridization for P. zoogleoformans

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Formalin-fixed, paraffin-embedded lung tissues were de-waxed and sequentially immersed in ethanol. The sections were washed with sterile PBS and incubated with lysozyme buffer (7 mg/mL) at 37 °C for 20 min. After being pretreated with hybridization buffer (0.9 NaCl, 20 mmol/L Tris-Cl (pH 8), and 0.1% (wt/vol) SDS) at 48 °C for 20 min, the sections were incubated with hybridization buffer containing 25% formamide and 0.2 μmol/L Alexa Fluor 594-labeled P. zoogleoformans probe (5′-TCCTTTACGGTTACGCACTTC-3′) in a dark humid chamber at 48 °C for 2 h. The sections were then washed with hybridization washing solution (0.02 mol/L NaCl, 20 mmol/L Tris-Cl (pH 8), 0.5 mol//L EDTA and 0.1% (wt/vol) SDS) for 25 min at 48 °C.^{60 (link),61 (link)} The sections were counterstained with DAPI (P36931, Thermo Fisher Scientific).

Meng X., Du L., Xu S., Zhou L., Chen B., Li Y., Chen C., Ye H., Zhang J., Tian G., Bai X., Dong T., Lin W., Sun M., Zhou K., Liu Y., Zhang W, & Duan S. (2024). Periodontitis exacerbates pulmonary hypertension by promoting IFNγ+ T cell infiltration in mice. International Journal of Oral Science, 16, 27.

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Cell Adhesion Assay on Substrates

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Substrates were housed in 6-well plates during the experiment. Cells were seeded by pipetting cell suspensions directly over the substrates. After 1hr, non-adhered cells were washed away. Cells were fixed with warmed 4% paraformaldehyde and the substrates were mounted onto glass slides using DAPI-infused mounting medium (ThermoFisher P-36931) 8 hours after seeding and later imaged.

Pushkarsky I., Tseng P., Warfe L., Black D., France B., Koziol-White C.J., Jester WF J.r., Trinh R.K., Lin J., Scumpia P.O., Morrison S.L., Panettieri RA J.r., Damoiseaux R, & Di Carlo D. (2018). Elastomeric sensor surfaces for high-throughput single-cell force biology. Nature biomedical engineering, 2(2), 124-137.

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Immunohistochemistry of Hippocampal Slices

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All immunohistochemical staining was performed on hippocampal slices used for the electrophysiology experiments. When the experimental recordings finished, slices (350 μm) were taken from the interface chamber and immediately drop fixed in 4% PFA for 4–24 h before being transferred to a 30% sucrose solution for >24 h. The slices were then re-sectioned using a freezing microtome to 70 μm and stored in a cryoprotective solution (25% glycerol, 30% ethylene glycol in PBS) at −20°C until required.
For immunostaining, sections were rinsed in PBS at room temperature then blocked for 2 h in 10% normal goat serum with 0.5% Triton X-100 then incubated in the primary antibody at 4°C for 24–48 h. Sections were then rinsed with PBS and incubated in secondary antibodies (1:1,000) for 2 h at room temperature. All antibodies were diluted in a carrier solution consisting of PBS with 1% BSA, 1% normal goat serum, and 0.5% Triton X-100. Sections were then rinsed, mounted on Superfrost glass slides, and cover slipped using prolong gold antifade mounting medium (Thermo Fisher Scientific; Cat# P-36931) and 1.5 mm cover glasses.

Mackenzie-Gray Scott C.A., Pelkey K.A., Caccavano A.P., Abebe D., Lai M., Black K.N., Brown N.D., Trevelyan A.J, & McBain C.J. (2022). Resilient Hippocampal Gamma Rhythmogenesis and Parvalbumin-Expressing Interneuron Function Before and After Plaque Burden in 5xFAD Alzheimer’s Disease Model. Frontiers in Synaptic Neuroscience, 14, 857608.

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Immunohistochemistry of Cryosectioned Tissues

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Tissues were fixed with 4% PFA (Santa-Cruz sc-281692) for 4–6 hours on a shaker at 4°C. They were then washed with PBS three times and perfused in 20% sucrose solution in PBS overnight at 4°C. They were subsequently mounted in the OCT embedding compound and frozen first at −20 and then at −80 °C. Tissue sections were prepared at 8 µm thickness using a cryostat and mounted onto gelatin-coated histological slides, which were stored at −80°C.
For immunostaining, slides were thawed to room temperature and fixed in pre-cold acetone for 10 minutes, followed by rehydration in PBS for 10 minutes. The slides were incubated in a blocking buffer (1% horse serum and 0.02% Tween 20 in PBS) for 1–2 hours at room temperature, followed by incubation with primary antibodies, which were diluted in an incubation buffer (1% horse serum, 0.02% Tween 20 in PBS), overnight at 4 °C. The slides were then washed three times with PBS and incubated with a secondary antibody [donkey anti-rabbit IgG H&L (DyLight® 550) preadsorbed (abcam ab96920)] in the incubation buffer for 1 hour at room temperature. After repeated washes the slides were mounted with an anti-fade mounting media containing DAPI (Thermo Fisher P36931">P36931) and visualized using a confocal microscope.

Xiao Q., Wu J., Wang W.J., Chen S., Zheng Y., Yu X., Meeth K., Sahraei M., Bothwell A.L., Chen L., Bosenberg M., Chen J., Sexl V., Sun L., Li L., Tang W, & Wu D. (2018). DKK2 imparts tumor immunity evasion through β-catenin-independent suppression of cytotoxic immune cell activation. Nature medicine, 24(3), 262-270.

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Visualizing Cell Adhesion and Differentiation

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All samples were washed with PBS to remove any unattached cells and fixed in situ with 4% paraformaldehyde for 10 min at room temperature. Then, cells were permeabilized for 5 min with PBS containing 0.1% TritonX-100 and unspecific binding blocked with 2% BSA in PBS for 1 h. To visualize focal adhesions (FAs) or differentiation, cells were treated with the three primary antibodies, including mouse anti-vinculin antibody (Sigma; V9131; 1:300), mouse anti-types I collagen (Col-I) antibody (Abcam; ab6308; 1:200), or mouse anti-α-SMA antibody (Sigma; A5228; 1:200) at 4°C overnight, followed by incubation with goat anti-mouse-FITC (Sigma; F0257; 1:100) at 37°C for 1 h. Subsequently, F-actin was stained with phalloidin-TRITC (Cytoskeleton; PHDR1; 100nM) and the nucleus with DAPI (Thermo; P36931; undiluted).
All specimens were then examined under a ﬂuorescence microscope (Carl Zeiss, Oberkochen, Germany) at low magnification (5× or 10×) or a laser scanning confocal microscopy (Nikon Co., Japan) at high magnification (40×). The fluorescent images were quantitatively analyzed using NIH ImageJ software.

Lei X., Liu B., Wu H., Wu X., Wang X.L., Song Y., Zhang S.S., Li J.Q., Bi L, & Pei G.X. (2020). The effect of fluid shear stress on fibroblasts and stem cells on plane and groove topographies. Cell Adhesion & Migration, 14(1), 12-23.

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Visualizing Neutrophil Extracellular Traps

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Neutrophils were seeded on coverslips at a density of 100,000 cells/coverslip and treated as described above. The NETs or the unstimulated neutrophils were fixed using 4% paraformaldehyde. NETs were visualized through co-localization of extracellular DNA (DAPI), neutrophil elastase (Neutrophil Elastase with Alexa Fluor 594, Catalog SC-53388AF594, Santa Cruz Biotechnology) and citrullinated Histone H3 (rabbit anti-human citrullinated anti-histone H3, Abcam Catalog AB5103 with donkey anti-rabbit Alexa 647, Thermo Fisher, Catalog P36931) using confocal microscopy as previously described [11 (link),26 (link)].

Linssen R.S., Sridhar A., Moreni G., van der Wel N.N., van Woensel J.B., Wolthers K.C, & Bem R.A. (2022). Neutrophil Extracellular Traps Do Not Induce Injury and Inflammation in Well-Differentiated RSV-Infected Airway Epithelium. Cells, 11(5), 785.

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Immunofluorescence Staining of FLAG-tagged Cells

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Cells were grown on chamber slides (Fisher, 12-565-7), fixed with 10% Formalin and permeabilized with 0.1% Triton X-100 in PBS. Slides were then incubated with primary antibody (rabbit anti-FLAG, Sigma F7425 at 1:1000) in 1% BSA (Jackson Immunoresearch, 001-000-161) for 1 hr at room temperature. Slides were washed 3x with PBS and incubated with secondary Alexa Fluor-conjugated antibodies (Jackson Immunoresearch, 625 ng/ml) in 1% BSA for 45 min at room temp. Slides were again washed 3x with PBS and mounted in ProLong Gold with DAPI (Thermo Fisher P36931).

Freund E.C., Sapiro A.L., Li Q., Linder S., Moresco J.J., Yates JR I.I.I, & Li J.B. (2020). Unbiased Identification of trans Regulators of ADAR and A-to-I RNA Editing. Cell reports, 31(7), 107656.

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