Ileal sections were incubated in antigen retrieval solution (Dako; H3300, 1/100) while being heated using a PickCell Electric Cooker. After cooling down, ileal sections were incubated with blocking buffer (goat serum; Sigma; S26‐100ML, 1/100) for 30 min at room temperature. Subsequently, sections were stained with primary antibody (rabbit anti‐lysozyme (Dako; EC 3.2.1.17, 1/500) or rabbit anti‐DCAMKL1 (Abcam; Ab31704, 1/500)) overnight at 4°C. After washing the slides with PBS, ileum sections were counterstained with DAPI (Thermofisher; D21490, 1/1,000), UEA‐1 Fluorescein (Vector laboratories; FL‐1061, 1/1,000), and WGA (ThermoFisher; W11261, 1/200) and incubated with secondary antibody goat anti‐rabbit Alexa Fluor 568 (Thermofisher; A11036, 1/1,000). After 1 h, slides were mounted and later imaged with Zeiss AxioScan (10×) and processed with Zen Blue (anti‐DCAMKL1 staining) or imaged with Zeiss LSM880 Airyscan (60×) and processed with Zen Black software (anti‐lysozyme staining).
S26 100ml
Manufactured by Merck Group
The S26-100ML is a laboratory equipment product manufactured by Merck Group. It is a 100 milliliter vessel designed for general laboratory use. The core function of this product is to provide a contained space for various laboratory processes and experiments.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 880 airyscan, by Zeiss (1 mentions)
Axioscan, by Zeiss (1 mentions)
H3300, by Agilent Technologies (1 mentions)
Alexa fluor 568, by Thermo Fisher Scientific (1 mentions)
Ab31704, by Abcam (1 mentions)
Rabbit anti lysozyme, by Agilent Technologies (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Cell Signaling Technology (1 mentions)
Sc 516141, by Santa Cruz Biotechnology (1 mentions)
Fluorescence microscope, by Olympus (1 mentions)
Dulbecco phosphate buffered saline (dpbs), by Thermo Fisher Scientific (1 mentions)
P1304mp, by Thermo Fisher Scientific (1 mentions)
Alexa fluor 488 goat anti rabbit igg secondary antibody, by Thermo Fisher Scientific (1 mentions)
Alexa 633, by Thermo Fisher Scientific (1 mentions)
L0651, by Merck Group (1 mentions)
Streptavidin alexa fluor 633 conjugate, by Thermo Fisher Scientific (1 mentions)
Anti gfap, by Thermo Fisher Scientific (1 mentions)
Eclipse ti2 e, by Nikon (1 mentions)
Anti ki67, by Abcam (1 mentions)
Anti tubb3, by Merck Group (1 mentions)
5 protocols using s26 100ml
1
Immunofluorescent Staining of Ileal Sections
2
Investigating RANK Expression in RAW 264.7 Cells
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RAW 264.7 cells were plated in a chambered coverslip (18 wells) at a density of 2 × 104 cells/well and separated into the following 4 groups: 1. No stimulation: cells were incubated in DMEM medium alone. 2. Stimulated: cells were cultured in DMEM medium + 20 ng/mL M-CSF and 20 ng/mL RANKL. 3. Experiment group: cells were incubated with Ps-Gos (0.00001–100 µg/mL) + 20 ng/mL M-CSF and 20 ng/mL RANKL. 4. Positive control: cells were treated with 30 µM D-pinitol + 20 ng/mL M-CSF and 20 ng/mL RANKL. After incubation for 24 h, cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100 (No. 39487. Cell Signaling Technology, Danvers, MA, USA). After blocking with 5% goat serum (S26-100ML, Sigma Aldrich), they were incubated with the primary anti-mouse RANK antibody (1: 500; sc-390655, Santa Cruz Biotechnology) at 4 °C overnight with shaking. Subsequently, the cells were incubated with a secondary antibody (1:200; No. sc-516141, Santa Cruz Biotechnology) for 3 h and counterstained with 4′,6-diamidino-2-phenylindole (DAPI) (No. 4083, Cell Signaling Technology, Danvers, MA, USA) for 5 min. Finally, mounting medium was carefully added to preserve the fluorescence signal, and photographs were taken with a fluorescence microscope (Olympus, Tokyo, Japan). The fluorescence intensity was analysed with ImageJ software.
3
SARS-CoV-2 Spike Protein Detection
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293F cells (transduced and control) were collected and washed with D-PBS (Gibco, 14190250). Cells were incubated with blocking buffer (10% goat serum, Sigma, S26-100 ML; 1% IgG free bovine serum albumin, Sigma, A0336-50 mL; D-PBS) for 1 h at room temperature. After incubation, blocking buffer was aspirated and cells were incubated for 1 h at room temperature with primary antibody against SARS-CoV-2 spike (1:200; Invitrogen, 703959) prepared in staining buffer (1% IgG free bovine serum albumin in D-PBS). After incubation, cells were washed three times with D-PBS followed by a 1 h incubation at room temperature in Alexa Fluor 488 Goat anti-Rabbit IgG secondary antibody (1:1000; Invitrogen, A11034). Cells were then washed twice with D-PBS. Following washes, cells were suspended in propidium iodide (Invitrogen, P1304MP) diluted 1:1000 in D-PBS as a cell viability dye. Following two washes in D-PBS cells were transferred to a 24-well plate for imaging. Immunofluorescent images were acquired using a Zeiss AxioObserver.Z1 microscope with an AxioCam 506 color camera and Zen 2 software.
4
Visualizing Oligodendrocyte Progenitor Cells
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Mice were perfused 21 days’ post-induction (dpi) with 4% paraformaldehyde (PF) in a phosphate buffer (PB). They were then post-fixed for over 2 h in the same solution and stored at 4 °C in PBS. Coronal vibratome sections (50 µm) were washed and permeabilized three times with 0.5% Triton X-100 (PBS-T), washed three times in 0.1% PBS-T, and blocked for 30 min at room temperature (RT) with 5% normal goat serum (NGS, S26-100ML: Merck-Millipore). Brain sections were incubated overnight at 4 °C with the following antibodies in 5% NGS and 0.1% PBS-T: rabbit anti-PDGFRα (1:300, 3174S: Cell Signaling) and biotinylated tomato lectin (TL, 1:50, L0651: Sigma-Aldrich). After washing the brain slices three times with 0.1% PBS-T, they were incubated for 2 h at RT with a secondary antibody coupled to Alexa 633 (1:1000, Invitrogen) or a Streptavidin–Alexa Fluor 633 conjugate (1:1000, S21375: Invitrogen Life Technologies (Carlsbad,. CA, USA ). Prior to visualization, they were washed 6 times in 0.1% PBS-T and then 1× PBS.
5
Immunofluorescence Staining of Cell Cultures
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For immunofluorescence staining cells were blocked in 10% normal goat serum (NGS) (#S26-100ML, Merck Millipore) and visualized using the following primary antibodies: anti-GFAP (#14-9892, Invitrogen, mouse IgG1, 1:500), anti-TUBB3 (#T2200, Sigma, rabbit IgG, 1:200), anti-Ki67 (#ab16667, Abcam, rabbit, 1:200). Fluorescent detection was achieved by species-specific fluorophore-linked secondary antibodies: Alexa 555 α-mouse (#CST4409, Cell Signaling, 1:500 and Alexa fluor 488 α-rabbit (#CST4412, Cell Signaling, 1:500). DAPI (#28718–90-3, Roth, 1:1000) was used to stain cell nuclei. A microscope (Eclipse Ti2-E, Nikon) was used for analyses.
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