Log In Sign up
The largest database of trusted experimental protocols

Hoechst 33342

Manufactured by AnaSpec
Visit Supplier
Sourced in United States

Hoechst 33342 is a fluorescent dye that binds to the minor groove of DNA. It can be used to stain nuclei and quantify DNA content in cells.

Automatically generated - may contain errors

Lab products found in correlation

Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (4 mentions) Cell culture and transfection reagents, by Thermo Fisher Scientific (3 mentions) Abt 737, by Abbott (3 mentions) Hydrocortisone, by Merck Group (3 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Prolong gold, by Thermo Fisher Scientific (2 mentions) Anti bcl 2, by BD (2 mentions) Anti gapdh, by Merck Group (2 mentions) Cytochrome c, by Merck Group (2 mentions) Anti cd31, by BD (2 mentions) Anti puma, by Merck Group (2 mentions) Anti bak, by Merck Group (2 mentions) Anti actin, by BD (2 mentions) Anti bim, by Merck Group (2 mentions) Anti bad, by Merck Group (2 mentions) Abt 263 plx 4032 gsk 110212, by Selleck Chemicals (2 mentions) Staurosporine cisplatin dacarbazine vinblastine, by Merck Group (2 mentions) Anti a1 fl 175, by BD (2 mentions) Anti mcl 1, by Rockland Immunochemicals (2 mentions) Anti smac, by Merck Group (2 mentions)

Spelling variants of this product

Hoechst (9 citations) Hoechst 33342 staining (2 citations)

32 protocols using hoechst 33342

1

Stem/progenitor cell culture protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols
Two different types of cells were used to model stem/progenitor cells for in vitro studies. Multipotent mouse C2C12 myoblasts (ATCC® CRL-1772™, Manassas, VA, US) were cultured in Dulbecco's modified Eagle's media (DMEM, Thermo Fisher Scientific Inc.) with 10% fetal bovine serum (FBS; Thermo Fisher Scientific Inc.) and 1% penicillin-streptomycin (PS, 10,000 U/mL, Fisher Scientific). Poietics™ human mesenchymal stem cells (hMSCs) (PT-2501, Lonza Walkersville Inc. MD) were cultured with mesenchymal stem cell growth medium (MSCGM™ BulletKit™, PT-3001, Lonza Walkersville Inc. MD). All the cells were kept at 37 °C, 5% CO2 in a humidified incubator. No mycoplasma contamination was found in cell cultures, which was monitored using a mycoplasma detection kit (Nanjing Vazyme Biotech Co., Ltd. China, MycoBlue, #D101-02) and Hoechst 33342 (Anaspec, USA) staining.
Zhang X., Li K., Wang C., Rao Y., Tuan R.S., Wang D.M, & Ker D.F. (2024). Facile and rapid fabrication of a novel 3D-printable, visible light-crosslinkable and bioactive polythiourethane for large-to-massive rotator cuff tendon repair. Bioactive Materials, 37, 439-458.
+ Open protocol
+ Expand
2

Immunofluorescence Staining of Frozen Tumor Sections

Check if the same lab product or an alternative is used in the 5 most similar protocols
Unfixed tumors were embedded in OCT (Tissue Tek). Frozen sections were cut to a thickness of 10μm on a Leica cryostat and mounted on SuperFrost Plus slides (Fisher). Slides were air-dried for 10min, then fixed for 10min with 4% formaldehyde, rinsed with PBS, permeabilized with 0.5% Triton X-100 in PBS for 15min, then blocked for 1h (5% normal donkey serum, 1% BSA, 0.3% Triton X-100 in PBS) and incubated in primary antibody diluted in blocking buffer at 4°C overnight. After washing with PBS, secondary antibodies, conjugated to Alexa 488, Alexa FITC, Alexa 568, DyLight 649, and Hoechst 33342 (83218, AnaSpec) were diluted in blocking buffer and incubated with the slides for 1hr at room temperature (RT). After washing, slides were mounted in ProLong Gold (Invitrogen). Imaging was performed on a Nikon Eclipse TiE Microscope or a Zeiss LSM780 Confocal Microscope. Images have been analyzed in NIS-Elements, Zen software or ImageJ. Antibodies for immunofluorescence were CD49f (1:200; Biolegend, 313618), SOX2 (1:1000; Abcam, Ab92494), pKi67 (1:1000; Novocastra, NCL-Ki67p), PITX1 (1:500; Novus, NBP1–88644), KLF4 (1:1000; R&D, AF3158), phospho Histone H3 (ser19) (1:1000; Millipore, 06–570), Filaggrin (1:1000; BioLegend, 905801) and Nidogen (1:1000; Santa Cruz, sc-33706).
Sastre-Perona A., Hoang-Phou S., Leitner M.C., Okuniewska M., Meehan S, & Schober M. (2019). De novo PITX1 expression controls bi-stable transcriptional circuits to govern self-renewal and differentiation in squamous cell carcinoma. Cell stem cell, 24(3), 390-404.e8.
+ Open protocol
+ Expand
3

Visualizing Influenza Virus-Host Interactions

Check if the same lab product or an alternative is used in the 5 most similar protocols
For immunofluorescence, 293T cells were transfected in 48-well plates with plasmids expressing V5-tagged hnRNP UL1, MECR, or cMECR and 48 h later infected with WSN PB2-FLAG (MOI, 3; 8 h). A549 cells stably expressing hnRNP UL1 or MECR were seeded on coverslips in 12-well plates and infected with WSN PB2-FLAG (MOI, 0.5; 8 h). Monolayers were fixed with 4% paraformaldehyde in PBS for 10 min, quenched and permeabilized with 0.1 M glycine + 0.1% Triton X-100 in PBS for 5 min, and blocked with 3% BSA in PBS for 30 min at room temperature. Primary and secondary antibodies were sequentially incubated for 1 h each at room temperature at 1 μg/ml in blocking buffer. DAPI was added to 293T cells during secondary antibody incubation. Coverslips were mounted in medium containing DAPI (Vector Laboratories, H-1200). Images were captured using a 20X objective on an EVOS FL Auto (Thermo Fisher) and processed in Adobe Photoshop CC.
For live cell imaging, 10X or 20X images were captured on an EVOS FL Auto 1 d posttransfection. To visualize nuclei, Hoechst 33342 (Anaspec AS-83218) was added 10 minutes prior to capture. GFP-positive cells were enumerated using ImageJ by automating the following commands: threshold (300); binary > watershed; analyze particles (size 100 to 500 px).
Baker S.F., Meistermann H., Tzouros M., Baker A., Golling S., Polster J.S., Ledwith M.P., Gitter A., Augustin A., Javanbakht H, & Mehle A. (2022). Alternative splicing liberates a cryptic cytoplasmic isoform of mitochondrial MECR that antagonizes influenza virus. PLOS Biology, 20(12), e3001934.
+ Open protocol
+ Expand
4

Apoptotic Signaling Pathway Analysis

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
All cell culture and transfection reagents were from Invitrogen; and standard reagents were from Sigma or Fisher Scientific. Drugs were from: ABT-737 (Abbott Pharmaceuticals), ABT-263/PLX-4032/GSK-110212 (Selleck), zVAD-fmk (Calbiochem), Hoechst 33342 (Anaspec), and staurosporine/cisplatin/dacarbazine/vinblastine (Sigma). Antibodies (clone): anti-actin (C4), anti-A1 (FL-175), anti-BCL-2 (100), anti-CD31 (BD Pharmingen #550274) anti-MCL-1 (Rockland), anti-BAD (C7), anti-BIM (22 (link)–40), anti-PUMA (CT; Sigma), anti-SMAC (H177), anti-BID (C20), anti-cytochrome c (7H8.2C12), anti-BAK (G23), anti-BAX (6A7 for IP; N20 for western blot), anti-BCL-xL (H5 for IP; S18 for western blot), anti-GAPDH (9B3), anti-HSP60 (B-9), anti-p44/p42 MAPK ERK1/2 (137F5), and anti-phospho p44/42 MAPK ERK1/2 (197G2). Caspase-8 cleaved mouse BID (C8-BID) was from R&D Systems. Full-length BAX was made as described (24 (link)). Human BCL-xLΔC, MCL-1ΔC, and PUMAβ were made as described (17 (link)). The human BIM BH3 domain peptide (> 98% purity, Abgent) was resuspended in anhydrous DMSO in a N2 environment, stored at −80°C, and thawed only once. All lipids for the LUVs studies were purchased from Avanti Polar Lipids. Statistical significance was evaluated by two tailed Student t test for p < 0.05.
Serasinghe M.N., Missert D.J., Asciolla J.J., Wieder S.Y., Podgrabinska S., Izadmehr S., Belbin G., Skobe M, & Chipuk J.E. (2014). Anti-apoptotic BCL-2 proteins govern cellular outcome following B-RAFV600E inhibition and can be targeted to reduce resistance. Oncogene, 34(7), 857-867.
+ Open protocol
+ Expand
5

Isolation and Analysis of Cancer Stem Cells

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
Flow cytometry was used to analyze the SP/CSCs as described previously [48 (link)]. Briefly, 1 × 106 cells/ml of 10% FBS containing DMEM were incubated with 5 μg/mL Hoechst 33342 (AnaSpec Inc., Fremont, CA, USA) for 60 min at 37 °C. CSCs were sorted by FACS analysis using LSR II Green (BD Biosciences). Verapamil (50 μM/ml), an ABC transporters inhibitor, was used as a control to identify CSCs.
Vengoji R., Macha M.A., Nimmakayala R.K., Rachagani S., Siddiqui J.A., Mallya K., Gorantla S., Jain M., Ponnusamy M.P., Batra S.K, & Shonka N. (2019). Afatinib and Temozolomide combination inhibits tumorigenesis by targeting EGFRvIII-cMet signaling in glioblastoma cells. Journal of Experimental & Clinical Cancer Research : CR, 38, 266.
+ Open protocol
+ Expand
6

Immunocytochemical Analysis of Cellular Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols
H460 cells were seeded in 96-well Black/clear plates (Nunc) followed by treatment on the next day with P7170 for 1 h. Cells were fixed with 3.7% PFA in 1× PBS at RT for 20 min, cell membranes permeabilized with 0.1% Triton X-100 for 90 sec, followed by blocking with 5% BSA (Sigma-Aldrich Cat #A7030) (w/v) in 1 × PBS for 2 h before immunostaining. Primary antibodies used were pS6 and p4EBP1 (Cell Signaling) at dilution (1:500) at RT for 1 h. Secondary Goat Anti-Rabbit antibody IgG (H + L) DyLight 549 Thermo Scientific Cat #35557) probed at 1:1000 dilution for 1 h. Nuclei were stained with Hoechst 33342 (AnaSpec Inc. Cat #83218). Plates were scanned on Cellomics Array Scan VTI HCS Reader.
Venkatesha V.A., Joshi A., Venkataraman M., Sonawane V., Bhatia D., Tannu P., Bose J., Choudhari S., Srivastava A., Pandey P.K., Lad V.J., Sangana R., Ahmed T., Damre A., Deore V., Sahu B., Kumar S., Sharma S, & Agarwal V.R. (2014). P7170, a novel inhibitor of mTORC1/mTORC2 and Activin receptor-like Kinase 1 (ALK1) inhibits the growth of non small cell lung cancer. Molecular Cancer, 13, 259.
+ Open protocol
+ Expand
7

Synthesis and Characterization of Lipid-PEG Conjugates

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
O-Stearoyl Mannose (M-C18), polyethylene
glycol 2000-hydrazone-C18 (PHC), and polyethylene glycol 2000-amide-C18
(PAC) were synthesized following our previously published methods.31 (link),34 Doxorubicin hydrochloride was from Fisher Scientific Co. (Pittsburgh,
PA). Zoledronic acid, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium
bromide (MTT), PLGA (752H), and poly-d-lysine were from Sigma-Aldrich
(St. Louis, MO). Mannose was from Tokyo Chemical Industry Co., Ltd.
(Portland, OR). Hematoxylin-eosin (H&E) and anti-CD31 antibody
were from Abcam (Cambridge, MA). Hoechst 33342 was from AnaSpec, Inc.
(Fremont, CA). The 5-bromo-2′-deoxyuridine (BrdU) and primary
BrdU monoclonal antibody were from BD Biosciences (San Jose, CA).
Anti-CD206, RM0029-11H3, and FITC-labeled Anti-CD206 antibody were
from Santa Cruz Biotechnology, Inc. (Dallas, TX). Solvents used in
chemical synthesis were of analytical grade.
Niu M., Naguib Y.W., Aldayel A.M., Shi Y.C., Hursting S.D., Hersh M.A, & Cui Z. (2014). Biodistribution and in Vivo Activities of Tumor-Associated Macrophage-Targeting Nanoparticles Incorporated with Doxorubicin. Molecular Pharmaceutics, 11(12), 4425-4436.
+ Open protocol
+ Expand
8

Antibody-Based Cytoskeleton Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
The following antibodies were used: mouse anti-RhoG (Santa Cruz, sc-1007), mouse anti-myc (9E10) (Santa Cruz, sc-40), mouse anti-vinculin (mouse) (Sigma, V9131), rabbit anti-vinculin (Thermo-Fisher, 700062); rabbit anti-phospho-paxillin (Y118) (Cell Signaling, 2541), rabbit anti-lamellipodin (Cell Signaling, 91138), rabbit anti-Myosin Light Chain and rabbit anti-phospho-Myosin Light Chain (Ser 19) (Cell Signaling, 3671, 3672) mouse anti-alpha tubulin (Sigma, T9026), rabbit anti-tubulin (Abcam, ab18207); Alexa Fluor-488 and Alexa Fluor-594 anti-mouse IgG and anti-rabbit IgG conjugated secondary antibodies and Alexa Fluor-488 and Alexa Fluor-594 Phalloidin (Life Technologies). HRP-conjugated anti-mouse, anti-rabbit and anti-goat secondary antibodies (Jackson Immunoresearch). Hoechst 33342 (AnaSpec Inc., 83218).
Fibronectin (a gift from Keith Burridge, UNC-Chapel Hill, Chapel Hill, NC) and collagen type I (Thermo Fisher, A1048301) were used at indicated concentrations to coat coverslips. The contractility inhibitors (−)-blebbistatin (EMD Millipore) and Y27632 (LC Laboratories) were used as indicated. Nocodazole (Sigma) was used as indicated below.
Zinn A., Goicoechea S.M., Kreider-Letterman G., Maity D., Awadia S., Cedeno-Rosario L., Chen Y, & Garcia-Mata R. (2019). The small GTPase RhoG regulates microtubule-mediated focal adhesion disassembly. Scientific Reports, 9, 5163.
+ Open protocol
+ Expand
9

Examining Apoptosis-Regulating Proteins in MEFs

Cited 2 times
Check if the same lab product or an alternative is used in the 5 most similar protocols
All cell culture and transfection reagents were from Invitrogen; and standard reagents were from Sigma or Fisher Scientific unless indicated. Drugs were from: ABT-737 (Abbott Pharmaceuticals), Hoechst 33342 (Anaspec); and β-ME, Cisplatin, DTT, JC-1, mDIVI-1, Paclitaxel, Tg, TMRE, and Tun (Sigma). Antibodies (clone): anti-actin (C4), anti-BCL-2 (100), anti-MCL-1 (Rockland), anti-BIM (22-40), anti-PUMA (CT; Sigma), anti-cytochrome c (7H8.2C12), anti-BAK (NT), anti-BAX (clone 6A7 for IP; clone N20 for western blot; clone Δ21 for trypsin studies), anti-BCL-xL (S18), anti-GAPDH (9B3), anti-Mfn1 (ABCAM, 57602), anti-Mfn2 (ABCAM, 56889), anti-HSP60 (B-9). BAX, BAXΔC, BAXS184A, BCL-xLΔC, N/C-BID, BIM-S were made as described (Chipuk et al., 2008 (link); Suzuki et al., 2000 (link); von Ahsen et al., 2000 (link)). The human BIM and PUMA BH3 domain peptides (Abgent) were resuspended in anhydrous DMSO. Knockout and Wt matched MEFs: Bak−/−, Bax−/−, Bak−/−Bax−/−, Bid−/−, Bid−/−Bim−/−, Puma−/−, Mfn1−/− and Mfn2−/− were obtained from Drs. Stanley Korsmeyer (Wei et al., 2001 (link)), Doug Green (Chipuk et al., 2008 (link)), Gerald Zambetti (Jeffers et al., 2003 (link)), and ATCC (Chen et al., 2003 (link)).
Renault T.T., Floros K.V., Elkholi R., Corrigan K.A., Kushnareva Y., Wieder S.Y., Lindtner C., Serasinghe M.N., Asciolla J.J., Buettner C., Newmeyer D.D, & Chipuk J.E. (2014). Outer mitochondrial membrane shape engages BAX α9 to initiate mitochondrial outer membrane permeabilization and apoptosis. Molecular cell, 57(1), 69-82.
+ Open protocol
+ Expand
10

Apoptotic Signaling Pathway Analysis

Cited 1 time
Check if the same lab product or an alternative is used in the 5 most similar protocols
All cell culture and transfection reagents were from Invitrogen; and standard reagents were from Sigma or Fisher Scientific. Drugs were from: ABT-737 (Abbott Pharmaceuticals), ABT-263/PLX-4032/GSK-110212 (Selleck), zVAD-fmk (Calbiochem), Hoechst 33342 (Anaspec), and staurosporine/cisplatin/dacarbazine/vinblastine (Sigma). Antibodies (clone): anti-actin (C4), anti-A1 (FL-175), anti-BCL-2 (100), anti-CD31 (BD Pharmingen #550274) anti-MCL-1 (Rockland), anti-BAD (C7), anti-BIM (22 (link)–40), anti-PUMA (CT; Sigma), anti-SMAC (H177), anti-BID (C20), anti-cytochrome c (7H8.2C12), anti-BAK (G23), anti-BAX (6A7 for IP; N20 for western blot), anti-BCL-xL (H5 for IP; S18 for western blot), anti-GAPDH (9B3), anti-HSP60 (B-9), anti-p44/p42 MAPK ERK1/2 (137F5), and anti-phospho p44/42 MAPK ERK1/2 (197G2). Caspase-8 cleaved mouse BID (C8-BID) was from R&D Systems. Full-length BAX was made as described (24 (link)). Human BCL-xLΔC, MCL-1ΔC, and PUMAβ were made as described (17 (link)). The human BIM BH3 domain peptide (> 98% purity, Abgent) was resuspended in anhydrous DMSO in a N2 environment, stored at −80°C, and thawed only once. All lipids for the LUVs studies were purchased from Avanti Polar Lipids. Statistical significance was evaluated by two tailed Student t test for p < 0.05.
Serasinghe M.N., Missert D.J., Asciolla J.J., Wieder S.Y., Podgrabinska S., Izadmehr S., Belbin G., Skobe M, & Chipuk J.E. (2014). Anti-apoptotic BCL-2 proteins govern cellular outcome following B-RAFV600E inhibition and can be targeted to reduce resistance. Oncogene, 34(7), 857-867.
+ 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?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!