D9663 10ml

Manufactured by Merck Group

D9663-10ML is a laboratory product offered by Merck Group. It is a 10 milliliter volume container. The core function of this product is to store and handle materials in a laboratory setting, but no further details about its intended use are provided.

Automatically generated - may contain errors

Lab products found in correlation

T8787 50ml, by Merck Group (2 mentions) P9416 50ml, by Merck Group (2 mentions) Bp1600 100, by Thermo Fisher Scientific (1 mentions) Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions) P21 waf1 cip1, by Cell Signaling Technology (1 mentions) P16ink4a, by Cell Signaling Technology (1 mentions) C10850, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

D9663 (66 citations)

4 protocols using d9663 10ml

Immunohistochemistry Staining Protocol

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

Tissue blocks were sectioned at 12−13 μm and were left to dry on a slide warmer overnight. Wax slides were dewaxed and rehydrated. Tissue sections were permeabilized with 0.5% TritonX-100 in PBS for 5 minutes. Tissue sections were blocked in 5% donkey serum (Sigma, catalog #D9663-10ML), 5% goat serum (Sigma, catalog #G9023-10ML), 3% bovine serum albumin (Fisher, catalog #BP1600-100) and 0.1% TritonX-100 for 1 hour at room temperature. Primary antibodies were diluted in block at concentrations listed in the Key Resources Table and incubated overnight at 4°C. All fluorophore-conjugated secondary antibodies were diluted in block at 1:500. EdU staining was performed according to manufacturer recommendations (Invitrogen, see Proliferation Studies). DAPI nuclear staining was performed according to manufacturer recommendations. Tissue sections were washed 5× with PBS for 15 minutes after/between antibody incubations. Images of sections were captured on a Zeiss Imager M2 AxioCam MRm microscope. For pneumonectomy studies, imaging was focused exclusively on the accessory lobe for all mice. For quantification of cells, at least 10 randomly selected images were counted per animal. Images were processed with ImageJ/FIJI (version 2.0.0, NIH).

Lechner A.J., Driver I.H., Lee J., Conroy C.M., Nagle A., Locksley R.M, & Rock J.R. (2017). Recruited monocytes and Th2 cytokine signaling promote lung regeneration following pneumonectomy. Cell stem cell, 21(1), 120-134.e7.

+ Open protocol

+ Expand

Neural Stem Cell and Neuron Immunostaining

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

Cells were fixed with 4% paraformaldehyde at 40°C for 30 min and then permeabilized nd blocked with 1x PBS containing Triton (0.5% v/v, 20 min). The blocking step was performed using normal donkey serum (NDS) in PBS (Sigma, Cat.# D9663-10 ML) for 1 h at 5% v/v and 2 h at 10% v/v prior to nestin and β-III-tubulin (TUJ-1) immunostaining, respectively. Samples were immunostained at 4°C overnight with different primary antibodies: goat anti-nestin (undifferentiated neural stem cell marker, Santa Cruz Biotech, Cat.# sc-21248) or mouse anti-TUJ-1 (immature neuron marker, Neuromics, Cat.# MO15013). Primary antibodies were diluted to 3 μg/mL in 0.1% v/v Triton, 1% v/v NDS in PBS. After 3 additional washes, samples were incubated with secondary antibodies: donkey anti-goat or goat anti-mouse (Santa Cruz Biotech, Cat.# sc-362265 and sc-362257, respectively) in 1% v/v NDS for nestin stained samples and in 8% v/v NDS for TUJ-1 stained samples. After a 1 h incubation, cells were washed with a Tween solution (0.1% v/v in PBS) 3 times for 5 min. At the end, SYTOX green was added during 15 min at 1 µM to stain nuclei, as for proliferation assays. The stained samples were visualized under fluorescent microscope.

Haddad T., Noel S., Liberelle B., El Ayoubi R., Ajji A, & De Crescenzo G. (2016). Fabrication and surface modification of poly lactic acid (PLA) scaffolds with epidermal growth factor for neural tissue engineering. Biomatter, 6(1), e1231276.

+ Open protocol

+ Expand

Immunostaining Mesenchymal Stem Cells

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

To fix and immunostain the MSC samples, each Ibidi slide was first washed with PBS +/+. 4% PFA (Thermo Fisher Scientific, 28908) in 1

\times

PBS +/+ (Gibco) was used as the fixative. After 5–10 min of incubation, the slides were washed with PBS. For staining, the MSCs were first blocked using a mixture of 2% donkey serum (Sigma-Aldrich, D9663-10ML) and 0.5% Triton X-100 (Sigma-Aldrich, T8787-50ML) for 30 min. After blocking, the slides were washed with PBS twice, and then incubated with the primary staining solution (0.5% BSA, 0.25% Triton X-100, and the primary antibody, see Supplementary Table S1). The slides were left in the staining solution for 30 min and then washed twice with 1

\times

PBS. Afterwards, the secondary staining solution (with NucBlue and the secondary antibody) was added for 30 min. We then washed the samples twice with PBS and added 0.1% Tween 20 (Sigma-Aldrich, P9416-50ML) for storage.

Imboden S., Liu X., Lee B.S., Payne M.C., Hsieh C.J, & Lin N.Y. (2021). Investigating heterogeneities of live mesenchymal stromal cells using AI-based label-free imaging. Scientific Reports, 11, 6728.

+ Open protocol

+ Expand

Immunostaining and SA-β-Gal Assay

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

All antibodies were purchased from Cell Signaling Technology (p16Ink4a, p21 Waf1/Cip1, p38 MAPK antibody, CD44). Fixation and immunostaining of samples was performed as previously described (Imboden et al., 2021 ). Briefly, samples were washed with 1X PBS+/+ (with calcium and magnesium) twice followed by fixing with 4% PFA (ThermoFisher Scientific, 28908). After 5 min of incubation, the samples were washed again with 1X PBS+/+ twice. For staining, a blocking buffer solution prepared of 2% donkey serum (Sigma-Aldrich, D9663-10ML) and 0.5% Triton X-100 (Sigma-Aldrich, T8787-50ML) was added to each sample and incubated for 30 min. After the second washing step with 1X PBS+/+ staining solution of primary antibodies was added and incubated overnight at 4°C. Samples are washed again with 1X PBS+/+ followed by incubation of 30 minutes with second staining solution (secondary antibodies + NucBlue). Lastly, samples were washed twice with 1X PBS+/+ and additionally, 0.1% Tween (Sigma-Aldrich, P9416-50ML) was added for storage. The SA-β-gal staining (CellEvent, Invitrogen, C10850) was conducted using the vendor’s protocol. In summary, cells were washed twice with 1X PBS+/+ , followed by fixing with 4% PFA for 30 min at room temperature. After washing the cells twice with 1X PBS+/+ , the sample was incubated with the SA-β-gal working solution for 2 h at room temperature.

Francis C.A., Obraztsova A.Y, & Tebo B.M. (2000). Dissimilatory Metal Reduction by the Facultative Anaerobe Pantoea agglomerans SP1. Applied and Environmental Microbiology, 66(2), 543-548.

+ 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!