Dapi dye

Manufactured by Vector Laboratories

Sourced in United States, United Kingdom

DAPI (4',6-diamidino-2-phenylindole) is a fluorescent dye that binds strongly to DNA. It can be used to stain cell nuclei for visualization and quantification purposes.

Automatically generated - may contain errors

Lab products found in correlation

Normal goat serum (ngs), by Cell Signaling Technology (3 mentions) Anti mouse lc3 a b, by Cell Signaling Technology (3 mentions) Fv1000, by Olympus (3 mentions) Vectashield hardset mounting medium, by Vector Laboratories (3 mentions) Vectashield medium, by Vector Laboratories (2 mentions) Anti lamp2, by Abcam (1 mentions) Anti eea1, by Abcam (1 mentions) Anti calnexin, by Abcam (1 mentions) Alexa 488, by Thermo Fisher Scientific (1 mentions) Ptm 301, by PTM Biolabs (1 mentions) Bx51 fluorescence microscope, by Olympus (1 mentions) Fluorescence microscope, by Zeiss (1 mentions) Tcs sp2 mp, by Leica (1 mentions) Fluorescence conjugated secondary antibody, by Thermo Fisher Scientific (1 mentions)

10 protocols using dapi dye

Immunofluorescence Imaging of Prion Protein Localization

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

Cells were grown on poly-L-lysine-coated coverslips for 24 hours before
fixation in 4% paraformaldehyde and washed with PBS prior to blocking in 1% FBS,
0.3% Triton X-100. Cells were incubated at 4 °C for
12 hours in blocking buffer with anti-PrP primary antibodies,
i.e. 3F4 and D18 (InPro Biotechnology) monoclonal antibodies. The
following day, cells were incubated for 1 hour with secondary antibodies
conjugated with AlexaFluor. For PrP^C cell surface detection,
cells were incubated at 4 °C for 15 min and probed with
3F4 antibody. Cells were permeabilized with 0.2% Triton-X and stained with
AlexaFlour-488 secondary antibody. To detect Thioflavin-S (ThS)-positive
aggregates, transfected and non-transfected cells were fixed with 4% PFA/4%
sucrose/1% Triton X-100 in PBS. For organelle markers, we used anti-Calnexin (ER
marker), anti-EEA1 (early endosome marker), anti-Tfn (recycling endosome
marker), anti-M6PR (late endosomes marker) and anti-LAMP2 (lysosomes marker)
purchased from Abcam. Nuclei were stained with DAPI dye (VECTOR Laboratories).
Images were acquired with a DMIR2 confocal microscope equipped with Leica
Confocal Software (Leica).

Giachin G., Mai P.T., Tran T.H., Salzano G., Benetti F., Migliorati V., Arcovito A., Longa S.D., Mancini G., D’Angelo P, & Legname G. (2015). The non-octarepeat copper binding site of the prion protein is a key regulator of prion conversion. Scientific Reports, 5, 15253.

+ Open protocol

+ Expand

Rice Histone Protein Analysis

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

Western blotting was performed as previously described (Liu et al. 2018 (link)). Immunofluorescence analysis was performed using the method described by Gong et al. (Gong et al. 2009 (link)). The rice histone proteins were separated electophoretically in denaturing gels by SDS-PAGE (5%/12%). The antibodies used in this study were rabbit pan anti-Kbu antibody (1:5000; PTM BioLabs, HangZhou China, PTM-301) and rabbit anti-H3 antibody (1:10,000; PTM BioLabs, HangZhou China, PTM-1001); the goat secondary anti-rabbit antibody is conjugated with Alexa 488 (Invitrogen, A11008). Chromosomes were counterstained with DAPI dye (Vector Laboratories, H-1200).

Liu S., Liu G., Cheng P., Xue C., Zhou Y., Chen X., Ye L., Qiao Z., Zhang T, & Gong Z. (2019). Genome-wide Profiling of Histone Lysine Butyrylation Reveals its Role in the Positive Regulation of Gene Transcription in Rice. Rice, 12, 86.

+ Open protocol

+ Expand

Immunohistochemical Analysis of Microglia and NLRP3 in Mouse Brains

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

Brains of every group mouse were perfusion by 0.9% saline solution and fixed in 4% paraformaldehyde PBS solution at 4°C overnight. After dehydrated in 30% sucrose solution for one week, brains were sectioned coronally with embedding them with OCT. Rabbit anti-Iba1 (1 : 500; Abcam, #ab178847) and rabbit polyclonal to anti-NLRP3 (1 : 100; BOSTER, BM4490) as primary antibodies were applied. Secondary antibody including goat anti-rabbit Alexa Fluor 488 (1 : 500, EARTHOX, San Francisco, # E031220-01) was employed after primary antibodies. Before being covered with coverslips, the slides were added the antiquenching solution with DAPI dye (VECTASHIELD, USA) for observation. Images were taken in 5-vision/section stochastically to count the immunoreactive cells under Olympus BX51 fluorescence microscope [15 (link)].

Du X., Xu Y., Chen S, & Fang M. (2020). Inhibited CSF1R Alleviates Ischemia Injury via Inhibition of Microglia M1 Polarization and NLRP3 Pathway. Neural Plasticity, 2020, 8825954.

+ Open protocol

+ Expand

Immunofluorescence Detection of E-cadherin and MCAM/CD146

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

E‐cadherin and MCAM/CD146 immunofluorescence detection was performed with cells grown on glass coverslips (IBD). Cells were washed three times in PBS, fixed in 2% paraformaldehyde, washed three times in PBS, and then incubated with PBS containing 2% BSA for 15 min prior to application of the primary anti‐E‐cadherin or anti‐MCAM/CD146 antibodies (1 : 200) for 2 h at room temperature. Subsequently, cells were incubated for 45 min with the secondary antibody goat anti‐mouse IgG coupled to Alexa‐488 Fluor. Negative controls were obtained by omitting primary antibodies. Finally, the cells were mounted in Vectashield medium containing DAPI Dye (Vector, Peterborough, UK) and examined using a fluorescence microscope (Zeiss, Jena, Germany).

Delaunay T., Deschamps L., Haddada M., Walker F., Soosaipillai A., Soualmia F., El Amri C., Diamandis E.P., Brattsand M., Magdolen V, & Darmoul D. (2017). Aberrant expression of kallikrein‐related peptidase 7 is correlated with human melanoma aggressiveness by stimulating cell migration and invasion. Molecular Oncology, 11(10), 1330-1347.

+ Open protocol

+ Expand

Quantifying Autophagy Markers in Cells

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

Cells were plated at low confluence in 6 well plates (50,000 cells/well). On day 2, cells were exposed to serum starvation (0% FBS), normal medium (10% FBS), or chloroquine (50 μM) for 24 hours. Medium was removed, cells were washed with PBS and treated with 4% paraformaldehyde/PBS for 20 minutes at room temperature, washed, then permeabilized with 0.1% Triton X-100 for 10 minutes. Cells were then blocked with 5% normal goat serum (Cell Signaling Technology) containing 0.3% Triton X-100 in PBS for 60 minutes. Diluted primary antibody, anti-mouse LC3 A/B (Cell Signaling Technology), was applied in blocking buffer overnight at 4° C. Alexa Fluor-555 secondary antibody diluted in 1% normal goat serum in PBS were added for 1 hour at ambient temperature. Cells were fixed using Vectashield hard set mounting medium containing DAPI dye (Vector Laboratories). Images were acquired using confocal microscopy (Olympus FV-1000) and overlaid using ImageJ (22 (link)).

Clark C.A., Gupta H.B., Sareddy G., Pandeswara S., Lao S., Yuan B., Drerup J.M., Padron A., Conejo-Garcia J., Murthy K., Liu Y., Turk M.J., Thedieck K., Hurez V., Li R., Vadlamudi R, & Curiel T.J. (2016). Tumor-intrinsic PD-L1 signals regulate cell growth, pathogenesis and autophagy in ovarian cancer and melanoma. Cancer research, 76(23), 6964-6974.

+ Open protocol

+ Expand

Autophagy Induction by Serum Starvation and Chloroquine

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

Cells were plated at low confluence in 6-well plates (50,000 cells/well). Onday2, cells were exposed to serum starvation (0% FBS), normal medium (10% FBS), or chloroquine (50 mmol/L) for 24 hours. Medium was removed, cells were washed with PBS and treated with 4% paraformaldehyde/PBS for 20 minutes at room temperature, washed, and then permeabilized with 0.1% Triton X-100 for 10 minutes. Cells were then blocked with 5% normal goatserum (Cell Signaling Technology) containing 0.3% Triton X-100 in PBS for 60 minutes. Diluted primary antibody, anti-mouse LC3 A/B (Cell Signaling Technology), was applied in blocking buffer overnight at 4 °C. Alexa Fluor-555 secondary antibody diluted in 1% normal goat serum in PBS were added for 1 hour at ambient temperature. Cells were fixed using Vectashield hard set mounting medium containing DAPI dye (Vector Laboratories). Images were acquired using confocal microscopy (Olympus FV-1000) and overlaid using ImageJ.

Tang D., Yao R., Zhao D., Zhou L., Wu Y., Yang Y., Sun Y., Lu L, & Gao W. (2018). Trichostatin A reverses the chemoresistance of lung cancer with high IGFBP2 expression through enhancing autophagy. Scientific Reports, 8, 3917.

+ Open protocol

+ Expand

Immunofluorescence Staining and Confocal Microscopy

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

Immunofluorescence staining and confocal microscopy were performed as previously described
[27 (link)]. Briefly, cells were seeded onto coverslips coated with poly-L-lysine and incubated overnight at 37°C. After washing, the cells were fixed with formaldehyde, permeabilized with a permeation buffer, and blocked with 1% FBS. After overnight incubation with primary antibodies, the coverslips were incubated with fluorescence-conjugated secondary antibodies (Molecular Probes, Invitrogen, Carlsbad, CA, USA). The coverslips were then mounted with mounting medium containing DAPI dye (Vector Laboratories, Burlingame, CA, USA), and the fluorescence was visualized using a confocal laser microscope (Leica TCS Sp2 MP).

Lu Y.C., Chang J.T., Liao C.T., Kang C.J., Huang S.F., Chen I.H., Huang C.C., Huang Y.C., Chen W.H., Tsai C.Y., Wang H.M., Yen T.C., You G.R., Chiang C.H, & Cheng A.J. (2014). OncomiR-196 promotes an invasive phenotype in oral cancer through the NME4-JNK-TIMP1-MMP signaling pathway. Molecular Cancer, 13, 218.

+ Open protocol

+ Expand

Autophagy Induction Assay in Cultured Cells

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

Cells were plated at low confluence in 12‐well plates (50 000 cells/well). On day2, cells were exposed to serum starvation (0% FBS), normal medium (10% FBS), or chloroquine (50 mmol/L) for 24 hours. Medium was removed, cells were washed with PBS and treated with 4% paraformaldehyde/PBS for 20 minutes at room temperature, washed and then permeabilized with 0.1% Triton X‐100 for 10 minutes. Cells were then blocked with 5% normal goatserum (Cell Signaling Technology) containing 0.3% Triton X‐100 in PBS for 60 minutes. Diluted primary antibody, antimouse LC3 A/B (Cell Signaling Technology), was applied in blocking buffer overnight at 4°C. Alexa Fluor‐555 secondary antibodies diluted in 1% normal goat serum in PBS were added for 1 hour at ambient temperature. Cells were fixed using Vectashield hard set mounting medium containing DAPI dye (Vector Laboratories). Images were acquired using confocal microscopy (Olympus FV‐1000) and overlaid using ImageJ.

Tang D., Zhao D., Wu Y., Yao R., Zhou L., Lu L., Gao W, & Sun Y. (2018). The miR‐3127‐5p/p‐STAT3 axis up‐regulates PD‐L1 inducing chemoresistance in non‐small‐cell lung cancer. Journal of Cellular and Molecular Medicine, 22(8), 3847-3856.

+ Open protocol

+ Expand

Immunofluorescence Imaging of Adherent Cells

Cited 1 time

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

Immunofluorescence staining and confocal microscopy were performed as previously described^{43 (link)}. Briefly, cells were seeded onto coverslips coated with poly-l-lysine and incubated overnight at 37 °C. After the cells were washed, they were fixed in formaldehyde, permeabilized, and blocked with 1% FBS. After an overnight incubation with the primary antibodies, the coverslips were incubated with fluorescence- conjugated secondary antibodies (Molecular Probes, Invitrogen, Carlsbad, CA, USA). The coverslips were then mounted with mounting medium containing DAPI dye (Vector Laboratories, Burlingame, CA, USA), and the fluorescence was visualized using a confocal laser microscope (Leica TCS Sp2 MP, Leica Microsystems, Wetzlar, Germany).

Lu Y.C., Cheng A.J., Lee L.Y., You G.R., Li Y.L., Chen H.Y, & Chang J.T. (2017). MiR-520b as a novel molecular target for suppressing stemness phenotype of head-neck cancer by inhibiting CD44. Scientific Reports, 7, 2042.

+ Open protocol

+ Expand

Immunofluorescence Analysis of KLK6, PAR1, and PAR2 in HT-29 Cells

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

HT 29 cells seeded on a coverslip and let grown over night until 70–80% confluency was reached. Cells were fixed in 2% paraformaldehyde, and permeabilized with 0.1% Triton X-100, then cells were blocked with 5% BSA in TBS for 15 min at room temperature prior to application of the primary anti-KLK6 (1:50), anti-PAR1 antibodies (WEDE15) (1:100) or anti-PAR2 antibodies (Sam11) (1:50) at room temperature. All washing steps were performed using PBS. Subsequently, cells were incubated for 45 min with the secondary antibody goat anti-mouse or anti rabbit IgG-coupled to Alexa-Fluor^TM 488. Negative controls were obtained by omitting primary antibodies.
We also performed immunofluorescence studies following incubation of HT-29 cells for 20 min at 37 °C with KLK6 (20 nmol/L). The coverslips were mounted in Vectashield medium containing DAPI Dye (Vector, Peterborough, UK) and examined using a fluorescence microscope (Zeiss, Jena, Germany) and by confocal microscopy analysis (LSM 510 Zeiss; (Carl Zeiss QEC GmbH, Germany). Magnification 630X).

Bouzid H., Soualmia F., Oikonomopoulou K., Soosaipillai A., Walker F., Louati K., Lo Dico R., Pocard M., El Amri C., Ignatenko N.A, & Darmoul D. (2022). Kallikrein-Related Peptidase 6 (KLK6) as a Contributor toward an Aggressive Cancer Cell Phenotype: A Potential Role in Colon Cancer Peritoneal Metastasis. Biomolecules, 12(7), 1003.

+ 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!