A11017

Manufactured by Thermo Fisher Scientific

Sourced in United States, Japan, Canada

The A11017 is a laboratory equipment product by Thermo Fisher Scientific. It is designed for use in various scientific and research applications. The core function of this product is to provide a specific capability required for laboratory work, but a detailed description cannot be provided while maintaining an unbiased and factual approach.

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Lab products found in correlation

A11008, by Thermo Fisher Scientific (6 mentions) A11012, by Thermo Fisher Scientific (5 mentions) A 11072, by Thermo Fisher Scientific (4 mentions) Mab1501, by Merck Group (3 mentions) Na931v, by GE Healthcare (3 mentions) Alexa fluor 488, by Thermo Fisher Scientific (3 mentions) Na934v, by GE Healthcare (2 mentions) A11070, by GE Healthcare (2 mentions) A11072, by GE Healthcare (2 mentions) A21434, by Thermo Fisher Scientific (2 mentions) Paraformaldehyde 4, by Thermo Fisher Scientific (2 mentions) R960 25, by Thermo Fisher Scientific (2 mentions) A 21245, by Thermo Fisher Scientific (2 mentions) Vectashield vector h 1000, by Vector Laboratories (2 mentions) A21445, by Thermo Fisher Scientific (2 mentions) 4 6 diamidino 2 phenylindole (dapi), by Roche (2 mentions) A11070, by Thermo Fisher Scientific (2 mentions) Ab201340, by Abcam (2 mentions) Sc 393871, by Santa Cruz Biotechnology (2 mentions) Fitc conjugated secondary antibody, by Thermo Fisher Scientific (2 mentions)

Close spelling variants of this product

AlexaFluor 488 Goat anti-mouse (A11017) (2 citations)

31 protocols using a11017

Immunostaining of Teased Nerve Fibers

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All primary antibodies used are listed as follows:
Rabbit anti-Crb3 (D2, kind gift from Dr A.Le Bivic, IBDML, France, 1:200)
Mouse anti-Moesin (610401, BD transduction, 1:100)
Mouse anti-AnkG (NB20, Calbiochem, 1:50)
Rabbit anti-Gliomedin (KR34, Koch, 1:1,000)
Mouse anti-KV1.2 (75-008, Antibodies incorporated 1:500)
Rabbit anti-pERM (3149, Cell Signaling, 1:100)
Rabbit anti-pan-Neurofascin (563, kind gift of Dr L. Goutebroze, Institut du Fer à Moulin, Paris, France, 1:1,000)
Rabbit anti-Nav1.8 (AP672-4.1, kind gift of SR Levinson, university of Colorado Health Center, USA, 1:200)
Rabbit anti- Caspr/paranodin (L51, kind gift from Dr L. Goutebroze, Institut du Fer à Moulin, Paris, France, 1:1,000)
Mouse anti-MBP (SMI94, Sternberger Monoclonal incorporated, 1:10,000)
Rabbit anti-Willin (Ex988-1, kind gift from M. Takeichi, RIKEN Center for Developmental Biology, Japan, 1:200)
Rabbit anti- YAP (4912, Cell Signaling, 1:200)
Secondary antibodies for teased fibres immunostaining were used at 1:1,000 dilutions. To visualize rabbit primary antibodies: Alexa Fluor donkey anti rabbit IgGs (H+L) 488, 594 or 647 (A21206, A21207, A31573 respectively, Molecular Probes) was used. To visualize mouse primary antibodies: Alexa Fluor F(ab')2 fragments of goat anti mouse IgG (H+L) 488, 594 or 647 (A11017, A11020, A21237 respectively, Molecular Probes) was used.

Fernando R.N., Cotter L., Perrin-Tricaud C., Berthelot J., Bartolami S., Pereira J.A., Gonzalez S., Suter U, & Tricaud N. (2016). Optimal myelin elongation relies on YAP activation by axonal growth and inhibition by Crb3/Hippo pathway. Nature Communications, 7, 12186.

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Immunoblotting of Tubulin Modifications

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The primary antibodies used in this study were: mouse monoclonal anti-tubulin acetylated 611B-1 (Sigma-Aldrich, T6793, 1/1000), mouse monoclonal anti-polyglutamylation modification of tubulin (AdipoGen, GT335, 1/2000), rat polyclonal anti-CD49f G0H3 (BD Pharmingen, 555734, 1/5000), rabbit polyclonal anti-SMO (Abcam, ab38686, 1/500), rabbit monoclonal anti-GLI1 EPR4523 (Abcam, ab134906, 1/1000 for WB) and (Abcam, ab49314, 1/1000 for IF) and mouse monoclonal anti-β-actin (Millipore, MAB1501, 1/20,000). Preliminary results (unpublished) were obtained using rabbit polyclonal anti-GLI3C 2438B, mouse monoclonal anti-Gli3 N 6F5 and rabbit polyclonal anti-Gli3 N 2676A (Genentech) obtained courtesy of FJ de Sauvage [38] (link). Secondary antibodies were: AlexaFluor 488 or 594 goat anti-mouse (Molecular Probes, A11017, A11072, 1/400), goat anti-rat (Molecular Probes, A11006, A11007, 1/400), goat anti-rabbit (Molecular Probes, A11070, A11072, 1/400), ECL HRP-linked anti-mouse (GE Healthcare, NA931 V, 1/3000) and anti-rabbit (NA934 V, 1/3000).

Sénicourt B., Boudjadi S., Carrier J.C, & Beaulieu J.F. (2016). Neoexpression of a functional primary cilium in colorectal cancer cells. Heliyon, 2(5), e00109.

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Immunofluorescence Staining of Tumor Cells

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Tumor cells (3 × 10⁴) were seeded in chamber slides and treated as specified. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.5% Triton X-100. Cells were washed with PBS, blocked with 5% BSA/PBS, followed by incubation with the primary antibody (overnight at 4°C or rt for 2 hr). After washing, cells were incubated with secondary antibodies (rt for 1 hr) conjugated with Alexa Fluor 488 or 594 (Molecular Probes, #A11017, #A11020, #A11070, #A11072), mounted and examined using a fluorescence microscope (Nikon TiE microscope with an XM10 camera).

Sutton M.N., Glazer S.E., Muzzioli R., Yang P., Gammon S.T, & Piwnica-Worms D. (2024). Dimerization of the 4Ig isoform of B7-H3 in tumor cells mediates enhanced proliferation and tumorigenic signaling. Communications Biology, 7, 21.

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DNA Fiber Assay for Replication Dynamics

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DNA fiber assays were performed as described previously (15 (link)). Briefly, cultured cells were incubated for 20 minutes with media containing CIdU (25μM) followed by PBS wash and incubation with media containing IdU (250μM). Cell suspensions were pipetted onto glass microscopy slides and lysed. Slides were raised to an angle of 30° in order to stretch DNA fibers along the slide. Immunostaining was then performed. CIdU was detected using anti BrdU (rat) primary antibody (Abcam ab6326 1:400) and anti-rat alexa fluorophore 555 (Invitrogen A21434 1:500) secondary. IdU was detected using anti BrdU (mouse) primary antibody (BD 347580 1:500) and anti-mouse alexa fluorophore 488 (Invitrogen A11017 1:500).

Carruthers R.D., Ahmed S.U., Ramachandran S., Strathdee K., Kurian K.M., Hedley A., Gomez-Roman N., Kalna G., Neilson M., Gilmour L., Stevenson K.H., Hammond E.M, & Chalmers A.J. (2018). Replication stress drives constitutive activation of the DNA damage response and radioresistance in glioblastoma stem-like cells. Cancer research, 78(17), 5060-5071.

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Multicolor Immunofluorescence Imaging of NCI-H1299 Cells

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NCI-H1299 cells were fixed in 4% (w/v) paraformaldehyde at room temperature for 30 min and permeablized in 0.5% (w/v) Triton X-100, followed by incubation with primary and secondary antibodies for 1 h at room temperature sequentially. Anti-HA serum (AH158; Beyotime Institute of Biotechnology, Haimen, China) with 1:200 was used for the control and Alexa 488-conjugated secondary antibody (1:500, A-11017; Invitrogen; Thermo Fisher Scientific, Inc.) were used to probe for the NS1 protein at room temperature for 1 h. Following protein staining, anti-cytochrome c monoclonal antibody (1:1,000, BD556432; BD Biosciences, Franklin Lakes, NJ, USA) and Alexa 555-conjugated secondary antibody (1:500, A-21427; Invitrogen; Thermo Fisher Scientific, Inc.) were utilized to probe the NCI-H1299 cellular morphology. Finally, 4′,6-diamidino-2-phenylindole (DAPI, 1:1,000, D1306; Invitrogen; Thermo Fisher Scientific, Inc.) was used to dye the cell nucleus at room temperature for 1 h. Triple-fluorescence stained cells were observed with a confocal microscope at a high-power magnification of ×100 (FV10-ASW, version 01.07.03.00; Olympus Corporation, Tokyo, Japan).

Bian Q., Lu J., Zhang L., Chi Y., Li Y, & Guo H. (2017). Highly pathogenic avian influenza A virus H5N1 non-structural protein 1 is associated with apoptotic activation of the intrinsic mitochondrial pathway. Experimental and Therapeutic Medicine, 14(5), 4041-4046.

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Quantifying Multinucleation via Immunofluorescence

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For quantification of multinucleation, cells were fixed using paraformaldehyde 4% (Alfa Aesar) for 20 min. Coverslips were then washed using TBS. Cells were permeabilized and blocked for ≥1 h in TBS containing 0.02% saponin and 2% BSA (TBS-saponin-BSA). F-actin was stained using 1/100 Texas red-X Phalloidin (T7471; Invitrogen) or 1/50 Alexa Fluor 647 Phalloidin (A22287; Invitrogen). dPLCXD-V5 was revealed by immunostaining using a monoclonal anti-V5 antibody (1/1,000; R960-25; Invitrogen) and a goat Alexa Fluor 488–conjugated secondary antibody anti-mouse (1/400; A11017; Invitrogen). Coverslips were mounted using Vectashield with Dapi (Vector Laboratories). To assess multinucleation, ≥300 cells (n > 300) were counted manually per condition per N individual experiment unless otherwise specified.
Representative images were treated using SoftWorx software (GE Healthcare), ImageJ software (National Institutes of Health), and Photoshop (Adobe).

Mondin V.E., Ben El Kadhi K., Cauvin C., Jackson-Crawford A., Bélanger E., Decelle B., Salomon R., Lowe M., Echard A, & Carréno S. (2019). PTEN reduces endosomal PtdIns(4,5)P2 in a phosphatase-independent manner via a PLC pathway. The Journal of Cell Biology, 218(7), 2198-2214.

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Immunohistochemical Profiling of Immune Cells

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Four-µm-thick tissue sections prepared from formalin-fixed and paraffin-embedded tissue samples were deparaffinized in xylene twice for 10 min each and rehydrated in a graded ethanol series for 10 min each. After blocking endogenous peroxidase by 3% hydrogen peroxide and antigen retrieval by 14 min of microwave heating in EDTA buffer (pH 7.4), the samples were incubated with primary antibodies against CD8 (1:400, ab199016, Abcam, Cambridge, UK), CD103 (1:200, ab129202, Abcam), and CD69 (1:200, bs-2499R, Bioss, Woburn, MA, USA) overnight at 4 °C, and then incubated with peroxidase-linked secondary antibody for 30 min at room temperature. The samples were immersed in 3, 3-diaminobenzidine (DAB) and counterstained with Mayer’s hematoxylin, followed by dehydration in a graded ethanol series for 10 min each. After mounting, the samples were subjected to microscopic observation. For fluorescence immunohistochemical staining for CD8 and CD103, Alexa 488 Anti-mouse (1:500, A11017, Invitrogen, Tokyo, Japan) or Alexa 647 Anti-Rabbit (1:500, A21245, Invitrogen) was used as the secondary antibody, and DAPI (D21490, FluoroPure™ grade, Invitrogen) was used for DNA staining.

Hashimoto M., Kuroda S., Kanaya N., Kadowaki D., Yoshida Y., Sakamoto M., Hamada Y., Sugimoto R., Yagi C., Ohtani T., Kumon K., Kakiuchi Y., Yasui K., Kikuchi S., Yoshida R., Tazawa H., Kagawa S., Yagi T., Urata Y, & Fujiwara T. (2024). Long-term activation of anti-tumor immunity in pancreatic cancer by a p53-expressing telomerase-specific oncolytic adenovirus. British Journal of Cancer, 130(7), 1187-1195.

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Immunofluorescence Assay for DNA Damage Response

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Cells grown on glass slides or coverslips were fixed with PTEMF (0.2% Triton X-100, 0.02 M PIPES (pH 6.8), 0.01 M EGTA, 1 mM MgCl₂, 4% formaldehyde). After blocking with 3% BSA, cells were incubated with primary antibodies according to suppliers’ instructions: CREST (Antibodies Incorporated, 15-234-0001), γH2aX (Millipore, 05-636), RPA70 (Abcam, ab79398). Secondary antibodies used were goat anti-mouse AlexaFluor 488 (A11017, Invitrogen), goat anti-rabbit AF594, AF488 (A11012, A11008, Invitrogen), and goat anti-human AF647 (109-606-088-JIR, Stratech or A21445, Invitrogen). DNA was stained with DAPI (Roche) and coverslips mounted in Vectashield (Vector H-1000, Vector Laboratories). EdU incorporation and staining was achieved using the Click-It kit (Life Technologies), following the manufacturer’s instructions.

Shaikh N., Mazzagatti A., De Angelis S., Johnson S.C., Bakker B., Spierings D.C., Wardenaar R., Maniati E., Wang J., Boemo M.A., Foijer F, & McClelland S.E. (2022). Replication stress generates distinctive landscapes of DNA copy number alterations and chromosome scale losses. Genome Biology, 23, 223.

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Immunofluorescence Assay for Cytoskeletal and Centromeric Proteins

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Cells grown on glass slides or coverslips were fixed with PTEMF (0.2% Triton X-100, 0.02 M PIPES [pH 6.8], 0.01 M EGTA, 1 mM MgCl₂, and 4% formaldehyde). After blocking with 3% BSA, cells were incubated with primary antibodies according to suppliers’ instructions: beta-tubulin (ab6046; Abcam), Centrin 3 (ab54531; Abcam), CREST (15-234-0001; Antibodies Incorporated), and CENP-E (ab5093; Abcam). Secondary antibodies used were goat anti-mouse Alexa Fluor 488 (A11017; Invitrogen), goat anti-rabbit AF594 and AF488 (A11012 and A11008; Invitrogen), and goat anti-human AF647 (109-606-088-JIR [Stratech] or A21445 [Invitrogen]). DNA was stained with DAPI (Roche), and coverslips were mounted in Vectashield (Vector H-1000; Vector Laboratories).

Worrall J.T., Tamura N., Mazzagatti A., Shaikh N., van Lingen T., Bakker B., Spierings D.C., Vladimirou E., Foijer F, & McClelland S.E. (2018). Non-random Mis-segregation of Human Chromosomes. Cell Reports, 23(11), 3366-3380.

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Immunofluorescence Staining of Midface Tissue

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Tissue sections were blocked for 1.5 h in 5% milk or 3% BSA dissolved in 0.1% Triton-X/PBS (PBSTx). Wild-type sections were used to examine protein expression of TFAP2A (3B5-supernatant, Santa Cruz Biotechnology; 1:25 dilution) and TFAP2B (2509S, Cell Signaling Technology; 1:25 dilution). E11.5 control and Tfap2^NCKO midface sections were stained with phosphorylated Histone H3 (9701S, Cell Signaling Technology; 1:400 dilution) to assess cell proliferation. After washing with PBSTx, samples were incubated with Alexa Fluor 488/568 secondary antibodies (A-11017, A-11070 and A-21069, Invitrogen; 1:500 dilution) for 1.5 h. We used DAPI to counterstain nuclei. Stains were imaged via Zen software on a Zeiss 700 LSM confocal microscope. Images were exported as Z-stacked, maximum intensity projection TIFF images.

Nguyen T.T., Mitchell J.M., Kiel M.D., Kenny C.P., Li H., Jones K.L., Cornell R.A., Williams T.J., Nichols J.T, & Van Otterloo E. (2024). TFAP2 paralogs regulate midfacial development in part through a conserved ALX genetic pathway. Development (Cambridge, England), 151(1), dev202095.

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