P36965

Manufactured by Thermo Fisher Scientific

Sourced in United States

The P36965 is a laboratory pipette that is designed for precise and accurate liquid handling. It features an ergonomic design and adjustable volume settings to accommodate a range of sample sizes. The pipette is suitable for a variety of laboratory applications.

Automatically generated - may contain errors

Lab products found in correlation

Lsm 880 confocal microscope, by Zeiss (2 mentions) Alexa fluor 488, by Thermo Fisher Scientific (2 mentions) Lsm 710 confocal, by Leica camera (1 mentions) Alexa fluor conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (1 mentions) Alexa fluor secondary antibody, by Thermo Fisher Scientific (1 mentions) Paraformaldehyde (pfa), by Merck Group (1 mentions) Tomm20, by Cell Signaling Technology (1 mentions)

5 protocols using p36965

Quantifying Intracellular Francisella Bacteria

Check if the same lab product or an alternative is used in the 5 most similar protocols

BMDMs were seeded on 8-well glass slides (Millicell EZ slide; #PEZGS0416) and were infected as described above. At the desired time points, cells were washed two times and fixed for 20 min at RT with 4% paraformaldehyde. Following fixation, cells were washed three times with PBST, and incubated with blocking buffer overnight at 4°C. Cells were stained for 1 h at RT with primary antibodies (identified below), then washed with PBST and incubated for 1 h at RT with the appropriate Alexa Fluor–conjugated secondary antibodies (identified below) (1:10,000 dilution; Invitrogen) were washed three times with PBST and 1X-DAPI (0.1μg/ml) was added to each well. After washing three times with PBST, mounting media (Invitrogen, #P36965) was added to each well and then coverslips added to the wells. Antibodies used were anti–F. tularensis LPS (1:1000 dilution; #MA1-21690; Invitrogen) to stain SCHU S4 and LVS, rabbit anti-GBP2 (1:500 dilution; 11854-1-AP; Proteintech) and anti-GBP5 (1:500 dilution; 13220-1-AP; Proteintech).
Coverslips were analyzed using Zeiss LSM710 Confocal and Leica Widefield Thunder microscopes at the magnification of ×63 or ×100, respectively. Total intracellular bacteria were quantified by an automated process using Fiji software (https://imagej.net/software/fiji/).

Mohammadi N., Lindgren H., Yamamoto M., Martin A., Henry T, & Sjöstedt A. (2021). Macrophages Demonstrate Guanylate-Binding Protein-Dependent and Bacterial Strain-Dependent Responses to Francisella tularensis. Frontiers in Cellular and Infection Microbiology, 11, 784101.

+ Open protocol

+ Expand

Immunofluorescence Staining of Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

For cell grown on coverslips, fixing and permeabilisation was performed in 4% paraformaldehyde (Sigma) and 0.1% Triton-100 (Sigma) before blocking in 1% Bovine Serum Albumin. The cells were then incubated with the primary antibodies overnight at 4 °C at a dilution of 10 µg/µl). pERM (Cell Signalling Technology; Antibody #3141), anti-integrin avβ6 (Abcam, ab97588). Alexafluor secondary antibodies (Invitrogen) were used at a 1/200 dilution. The coverslips were mounted on glass slides using mounting medium with DAPI (Invitrogen P36965). For cells grown in Matrigel, following fixing and permeabilisation as detailed above non-specific staining was blocked using cells were blocked using IF Buffer (7.7 mM NaN3, 0.1% bovine serum albumin, 0.2% Triton X-100, 0.05% Tween-20 in PBS) + 10% goat serum. Cells were stained using Phalloidin for F-actin (Sigma, P1951) and mounted using DAPI (Invitrogen).
For all experiments three technical and three biological replicates were performed.

Ran R., Harrison H., Syamimi Ariffin N., Ayub R., Pegg H.J., Deng W., Mastro A., Ottewell P.D., Mason S.M., Blyth K., Holen I, & Shore P. (2020). A role for CBFβ in maintaining the metastatic phenotype of breast cancer cells. Oncogene, 39(12), 2624-2637.

+ Open protocol

+ Expand

Immunostaining of Hippocampal Neurons

Check if the same lab product or an alternative is used in the 5 most similar protocols

PFA (4%) was used for fixation of primary Hippocampal neurons. Neurons were then permeabilized with 0.5 % Triton and blocked with 5% bovine serum albumin (BSA) for 1 hour at room temperature (RT), and incubated at room temperature for 2 hours or overnight at 4°C with the following primary antibodies: MPC1 antibody (1:500, Sigma HPA045119) guinea pig vGLUT1 (1:500, Sigma AB5905); TOMM20 (1:500, Cell Signaling Technology 42406S). Coverslips were then incubated with the following secondary antibodies: anti-rabbit Alexa-fluor568 (1:500, Thermo FisherA21428) and anti-mouse Alexa-fluor488 (1:500, Thermo Fisher A11059) for 1 hour at RT and mounted with anti-fade mounting media (Thermo Fisher P36965), and kept at 4°C until imaged. Confocal images were collected on a Zeiss LSM 880 Confocal Microscope at the Washington University Center for Cellular Imaging which was purchased with support from the Office of Research Infrastructure Programs (ORIP), as part of the NIH Office of the Director under grant OD021629.

Tiwari A., Myeong J., Hashemiaghdam A., Zhang H., Niu X., Laramie M.A., Stunault M.I., Sponagel J., Patti G., Shriver L., Klyachko V, & Ashrafi G. (2024). Mitochondrial pyruvate transport regulates presynaptic metabolism and neurotransmission. bioRxiv.

+ Open protocol

+ Expand

Immunofluorescence Microscopy Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunofluorescence was performed as previously described in Nagashima et al. [80 (link)]. Cells were plated on coverslips at a density of 20,000-25,000 cells per well in a 24-well plate. The next day, the cells were fixed with warmed paraformaldehyde (PFA) in PBS (pH 7.4) for 15 min at 37 °C, treated with 50 mM NH₄Cl₂ for 10 min at RT to quench autofluorescence and incubated with 0.1% Triton for 10 min at RT. Blocking was conducted using 10% FBS-PBS for 20 min at RT. Primary antibodies were incubated in humidity boxes overnight at 4 °C in 5% FBS-PBS. The next day, the coverslips were washed three times in 5% FBS-PBS and incubated with secondary antibodies for 1 h at RT in 5% FBS-PBS. Cells were washed three times in 1x PBS, incubated with Hoechst if needed, and then rinsed in ddH2O before mounting on slides using Dako Mounting media (S3023) or Prolong gold Diamond (Thermo, P36965) for spinning disk confocal or structured illumination microscopy (SIM), respectively.

Wilson E.L., Yu Y., Leal N.S., Woodward J.A., Patikas N., Morris J.L., Field S.F., Plumbly W., Paupe V., Chowdhury S.R., Antrobus R., Lindop G.E., Adia Y.M., Loh S.H., Prudent J., Martins L.M, & Metzakopian E. (2024). Genome-wide CRISPR/Cas9 screen shows that loss of GET4 increases mitochondria-endoplasmic reticulum contact sites and is neuroprotective. Cell Death & Disease, 15(3), 203.

+ Open protocol

+ Expand

Immunostaining of Dnmt3a2/3L Mutant Brains

Check if the same lab product or an alternative is used in the 5 most similar protocols

Brains from Dnmt3a2/3L^Dat^/wt mice and Dnmt3a2/3L^wt^/wt littermate controls were harvested, and embedded in OCT. Brain sections (30 μm) were immunostained with primary antibody overnight at 4 °C, washed PBS (6 × 5 min), then incubated with secondary antibody for 2 h at room temperature, washed with PBS (6 × 5 min), and mounted using mounting gel (Thermo Fisher, P36965, Waltham, MA, USA). The next day, the slices were visualized under a fluorescence microscope. For DAB staining, after washing, the brain sections were incubated with the secondary antibody, and the HRP reagent for 1h. Reactions were visualized by developing DAB substrates and were visualized under a light microscope.

Cui D., Mesaros A., Burdeos G., Voigt I., Giavalisco P., Hinze Y., Purrio M., Neumaier B., Drzezga A., Obata Y., Endepols H, & Xu X. (2020). Dnmt3a2/Dnmt3L Overexpression in the Dopaminergic System of Mice Increases Exercise Behavior through Signaling Changes in the Hypothalamus. International Journal of Molecular Sciences, 21(17), 6297.

+ Open protocol

+ Expand

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?

Revolutionizing how scientists
search and build protocols!