Alexa fluor 568 goat anti mouse antibody

Manufactured by Thermo Fisher Scientific

Sourced in United States

Alexa Fluor 568 goat anti-mouse antibody is a secondary antibody conjugated with the Alexa Fluor 568 fluorescent dye. It is designed to bind to and detect mouse primary antibodies in various immunological applications, such as immunofluorescence, flow cytometry, and Western blotting.

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Lab products found in correlation

Alexa fluor 488 goat anti rabbit antibody, by Thermo Fisher Scientific (4 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (2 mentions) Lsm 880 confocal microscope, by Zeiss (2 mentions) Anti human mpo, by Abcam (1 mentions) Fibronectin, by Merck Group (1 mentions) Hoechst 33342, by Thermo Fisher Scientific (1 mentions) Decma 1, by Abcam (1 mentions) Mouse anti paxillin antibody, by BD (1 mentions) Sc 373717, by Santa Cruz Biotechnology (1 mentions) Vectashield, by Vector Laboratories (1 mentions) Cryostat microtome, by Leica (1 mentions) Ab109186, by Abcam (1 mentions) Ab20670, by Abcam (1 mentions) Alexa fluor 488 goat anti rabbit, by Thermo Fisher Scientific (1 mentions) 4 6 diamidine 2 phenylindole dihydrochloride dapi, by Roche (1 mentions) Tcs sp5, by Leica (1 mentions) Bovine serum albumin (bsa), by Rockland Immunochemicals (1 mentions) Reducing agent, by Bio-Rad (1 mentions) Carbo free blocking solution, by Vector Laboratories (1 mentions) Dylight 649 tomato lectin, by Vector Laboratories (1 mentions)

Spelling variants of this product

Alexa Fluor 568 (1190 citations) Alexa 568 (337 citations) Alexa Fluor 568 goat anti-mouse (158 citations) Anti-mouse Alexa Fluor 568 (44 citations) Goat anti-mouse Alexa 568 (42 citations) Alexa-568 anti-mouse (9 citations) Anti-goat Alexa-Fluor 568 (6 citations) Alexa Fluor 568 goat (5 citations) Anti-mouse Alexa Fluor (4 citations) Alexa Fluor 568 anti-mouse antibody (2 citations)

13 protocols using alexa fluor 568 goat anti mouse antibody

Quantification of NET Formation

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NET formation was monitored by the presence of extracellular DNA covered with MPO. Cover glasses (CS) were coated with fibronectin (2.5μg/ml, Sigma-Aldrich) for 1h. 3x10⁴ freshly isolated PMNs were resuspended in 30μl of RPMIΔ (RPMI 1640 GlutaMAX without supplements), seeded on CS and allowed to adhere for 20min. 20μl of medium containing either medium alone, DMSO, 100nM PMA or different amounts of HIV-1 viral particles were added to the CS and cells were incubated for 3 or 24h at 37°C, 5% CO₂. Cells were then fixed with 3% PFA for 90min at room temperature, permeabilized with 0.1% Triton X-100 and stained with anti-human MPO (1:100, clone 2C7, Abcam) followed by Alexa Fluor 568 goat anti-mouse antibody (1:2000, Life Technologies). Cells were additionally stained with Hoechst 33342 (1:10000, Invitrogen).
Quantification of NET formation was performed as follows: 5 random sections on the CS were imaged. Cells were divided in three categories: lobular nucleus, round nucleus and NET forming cell. The cells were attributed to one of the categories, counted manually and the percentage was calculated. NET formation in PMA treated samples was artificially set to 100% since the entire CS was covered with NETs which did not allow distinction.

Reif T., Dyckhoff G., Hohenberger R., Kolbe C.C., Gruell H., Klein F., Latz E., Stolp B, & Fackler O.T. (2021). Contact-dependent inhibition of HIV-1 replication in ex vivo human tonsil cultures by polymorphonuclear neutrophils. Cell Reports Medicine, 2(6), 100317.

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E-cadherin Perturbation and Focal Adhesion Visualization

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In experiments where antibodies were used to perturb E-cadherin-mediated cell–cell adhesions, MCF10A-E-cadherin-GFP cells were treated with 3 µg/ml of neutral, control antibody (76D5; gift of BM Gumbiner (Petrova et al., 2012 (link))), or E-cadherin-blocking antibody (DECMA-1; Abcam) for at least 2 hr prior to TFM measurements. As an additional control, cells were also imaged and measured pre-treatment.
In immunostaining experiments to visualize focal adhesions in MCF10A-E-cadherin-GFP cell clusters, a purified mouse anti-paxillin antibody (BD Biosciences, San Jose, CA) was used as the primary antibody and the Alexa Fluor 568 goat anti-mouse antibody (Life Technologies, Grand Island, NY) was used as the secondary antibody.

Ng M.R., Besser A., Brugge J.S, & Danuser G. (2014). Mapping the dynamics of force transduction at cell–cell junctions of epithelial clusters. eLife, 3, e03282.

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Immunohistochemical Analysis of Spermatogenesis

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Frozen testis sections (10 μm) from 3- to 4-month-old Bscl2^+/+ and Bscl2^−/− males (N=4) were used for detecting the expression of vimentin, PRM1, and PRM2 following the procedure as described previously.^{49 (link)} Sections were incubated with vimentin antibody (1:50 dilution, sc-373717, Santa Cruz Biotechnology) or PRM1 antibody (1:100 dilution, 21 μg/ml, Hup1N, Briar Patch Biosciences, Livermore, CA, USA), or PRM2 antibody (1 : 100 dilution, 21 μg/ml, Hup2B, Briar Patch Biosciences) overnight at 4 °C. Vimentin, PRM1, and PRM2 signals were detected by incubating with Alexa Fluor 568 goat anti-mouse antibody (1 : 200 dilution, Life Technologies) for 30–60 min at room temperature. Testis sections were counterstained and mounted in DAPI-containing Vectashield (Vector Laboratories). Two types of negative control were used: sections of 3-month-old Bscl2^+/+ males incubated with mouse IgG (1 : 100, Santa Cruz Biotechnology) or without any primary antibody. All other procedures were the same as described above. In addition, PRM1 and PRM2 were also detected in the sperm spreads from Bscl2^+/+ and Bscl2^−/− males (4 months old, N=3) using the same condition as for testis sections.

El Zowalaty A.E., Baumann C., Li R., Chen W., De La Fuente R, & Ye X. (2015). Seipin deficiency increases chromocenter fragmentation and disrupts acrosome formation leading to male infertility. Cell Death & Disease, 6(7), e1817-.

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Immunostaining Protocol for Embryoid Bodies

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Immunostaining of EBs was carried out as previously described (22 (link)). Briefly, EBs were fixed and dehydrated with 4% Paraformaldehyde (PFA) and followed by sequential Sucrose incubation. After cryo-embedding, 7 μm-thickness of sectioning was performed with a cryostat microtome (Leica, Vetzlar, Germany). For immunostaining, EB sections were fixed with 4% PFA for 15 min, permeabilized with 0.5% Trinon X-100 for 5 min, blocked with 3% BSA for 30 min and probed with the following antibodies; Nkx6.1 (F55A12, DSHB, Iowa, USA, 1：300), Olig2 (ab109186, Abcam, UK, 1：300), Islet1 (Isl1) (ab20670, Abcam, 1：300) or Hb9 (PA5-23407, Invitrogen, Massachu-setts, USA, 1：300). After over-night incubation with primary antibodies at 4℃, cells were labeled with appropriate secondary antibodies (Alexa Fluor 488 goat anti-rabbit or Alexa Fluor 568 goat anti-mouse antibody (Life technologies, California, USA, 1：300). Cells were counterstained with 4’,6-diamidine-2-phenylindole dihydrochloride (DAPI, Roche) to show the nuclei. Confocal images were obtained with a LSM-710 Meta confocal fluorescence microscope (Carl Zeiss, Oberhochen, Germany).

Kim H., Lee S.Y., Jeong H.J., Kang J.S., Cho H, & Leem Y.E. (2020). Cdo Is Required for Efficient Motor Neuron Generation of Embryonic Stem Cells. International Journal of Stem Cells, 13(3), 342-352.

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Immunostaining of Drosophila Eye-Antennal Discs

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Eye-antennal discs from wandering third instar larvae were dissected, and fixed in 4% paraformaldehyde in Phosphate-Buffered Saline (PBS), and stained following the protocol. The primary antibodies used were mouse anti-Chaoptin (MAb24B10) (1:100, DSHB #24B10, Iowa, IA, USA), mouse anti-22C10 (1:100, DSHB #22C10, Iowa, IA, USA) and rabbit anti-GFP (1:1000, ThermoFisher Scientific #A11122, Waltham, MA, USA), while the secondary antibody were Alexa Fluor 568 goat anti-mouse antibody (1:1000, ThermoFisher Scientific #A11031, Waltham, MA, USA) or Alexa Fluor 488 goat anti-rabbit antibody (1:1000, ThermoFisher Scientific # A11008, Waltham, MA, USA).
The brains of three-to-five-day-old mtt-Gal4 > UAS-mCD8::GFP flies were dissected with their retina preserved. At first, the whole flies were fixed in 4% formaldehyde at 25 °C for 2 h. Then, the dissected brains were washed in PBT (PBS containing 0.2% Triton X-100) and blocked in 5% goat serum in PBT (PBST) for 30 min at room temperature. The primary antibodies were used to incubate the samples overnight at 4 °C. Finally, samples were washed three times with PBT and incubated overnight with Alexa Fluor 488-labeled goat anti-rabbit or Alexa Fluor 568-labeled goat anti-mouse secondary antibody overnight at 4 °C. Images of mounted samples were captured under confocal microscope (Leica TCS SP5, Wetzlar, Germany).

Chen W., Zhong W., Yu L., Lin X., Xie J, & Liu Z. (2024). A Drosophila Model Reveals the Potential Role for mtt in Retinal Disease. International Journal of Molecular Sciences, 25(2), 899.

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Insulin Signaling Pathway Analysis

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Human insulin (Humulin R) was purchased from Eli Lily. FITC-insulin was purchased from Sigma-Aldrich. Vectastain Universal Elite ABC kit, Carbo-Free blocking solution, and DyLight 649-tomato lectin were purchased from Vector Laboratories. Bovine serum albumin (BSA) was purchased from Rockland Immunochemicals. Phospho-insulin receptor beta (Y1185) antibody was purchased from Bioss (#bs-5453R). Rabbit anti-insulin antibody and horseradish peroxidase conjugated anti-rabbit IgG antibody was purchased from Cell Signaling Technology. Neuro-Chrom pan-neuronal antibody was purchased from Millipore (#ABN2300). Phosphatase inhibitor cocktail was purchased from Research Products International. Complete mini protease inhibitor cocktail was purchased from Roche. SuperBlock blocking buffer, 4,6-diamidino-2-phenylindole (DAPI), IP lysis buffer, ProLong Diamond, and Alexa Fluor 568 goat anti-mouse antibody were purchased from Thermo Fisher. Any KD TGX gels, XT sample buffer, reducing agent, and Clarity Western ECL reagent were purchased from Bio-Rad. All other reagents were purchased from Sigma-Aldrich unless noted.

Lochhead J.J., Kellohen K.L., Ronaldson P.T, & Davis T.P. (2019). Distribution of insulin in trigeminal nerve and brain after intranasal administration. Scientific Reports, 9, 2621.

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Immunofluorescent Laminin Mapping of Fetal Hearts

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Human fetal hearts (gestational weeks 6–10, n = 3) were embedded in OCT, snap-frozen and thereafter cryo-sectioned and stained for anti-laminin α2 (MAB1922, clone 5H2, Millipore), anti-laminin α5 (MAB1924, clone 4C7, Millipore), or anti-laminin γ1 (MAB1920, clone 2 × 10⁸, Millipore) and visualized by a secondary Alexa Fluor 568 goat anti-mouse antibody (A11031, Thermo Fischer Scientific). The cell nuclei were counterstained with DAPI.

Månsson-Broberg A., Rodin S., Bulatovic I., Ibarra C., Löfling M., Genead R., Wärdell E., Felldin U., Granath C., Alici E., Le Blanc K., Smith C.I., Salašová A., Westgren M., Sundström E., Uhlén P., Arenas E., Sylvén C., Tryggvason K., Corbascio M., Simonson O.E., Österholm C, & Grinnemo K.H. (2016). Wnt/β-Catenin Stimulation and Laminins Support Cardiovascular Cell Progenitor Expansion from Human Fetal Cardiac Mesenchymal Stromal Cells. Stem Cell Reports, 6(4), 607-617.

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Immunostaining of Cartilage Matrix Proteins

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The deparaffinized paraffin sections were revitalized with antigen-retrieval reagent (Immunoactive; Matsunami Glass Ind, Osaka, Japan) at 60°C for 16 h or by Proteinase K (Dako) and blocked in 5% goat serum with 1% bovine serum albumin (BSA, FUJIFILM WAKO). After blocking, the samples were incubated with anti-TENOMODULIN antibody (1:200; ab203676, Abcam), anti-PRO COLLAGEN TYPE I antibody (DSHB, SP1.D8, 1:100), and anti-COLLAGEN TYPE II antibody (DSHB, CII C1, 1:500) overnight at 4°C. The sections were then incubated with Alexa Fluor 568 goat anti-rabbit antibody (1:500; Thermo Fisher Scientific) or Alexa Fluor 568 goat anti-mouse antibody (1:500; Thermo Fisher Scientific) for 1 h. Nuclei were stained using 4′,6-diamidino-2-phenylindole (DAPI) (DOJINDO, Japan).

Omoto T., Yimiti D., Sanada Y., Toriyama M., Ding C., Hayashi Y., Ikuta Y., Nakasa T., Ishikawa M., Sano M., Lee M., Akimoto T., Shukunami C., Miyaki S, & Adachi N. (2022). Tendon-Specific Dicer Deficient Mice Exhibit Hypoplastic Tendon Through the Downregulation of Tendon-Related Genes and MicroRNAs. Frontiers in Cell and Developmental Biology, 10, 898428.

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Assessing Protein Glutathionylation in Rotenone-Treated HEK293T Cells

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HEK293T cells treated with rotenone for 68 h and HEK293T NDUFA11-deficient cells were treated for the last 24 h with NAC, and then protein glutathionylation was analyzed in these cells and in untreated HEK293T cells. On the day of the experiment, cells were collected, washed twice with PBS, and fixed with 3.7% formaldehyde for 10 min at 4°C. Next, cells were washed twice with PBS, permeabilized for 5 min in 0.1% Triton X-100 solution in PBS, washed again with PBS, and incubated for 30 min in 10% FBS in PBS. After that, cells were incubated for 1 h at room temperature with an anti-glutathione antibody (1:250; Abcam; catalogue no. ab19534). After two washes with PBS, the cells were incubated for 1 h with Alexa Fluor 568 goat anti-mouse antibody (1:250; Invitrogen; catalogue no. A-11031) and then washed twice, resuspended in ice-cold PBS, and analyzed using a flow cytometer (BD LSRFortessa).

Samluk L., Ostapczuk P, & Dziembowska M. (2022). Long-term mitochondrial stress induces early steps of Tau aggregation by increasing reactive oxygen species levels and affecting cellular proteostasis. Molecular Biology of the Cell, 33(8), ar67.

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Immunofluorescence Staining of Spinal Cord

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Spinal cord sections were prepared as described above. For tissue immunofluorescence staining, sections were blocked with blocking buffer (1× PBS/5% BSA/0.3% Triton^TM X-100) for 1 hour at room temperature. Sections were incubated with mouse anti-Iba1 antibody (1:100, Abcam, Cambridge, UK, Cat# ab283319, RRID: AB_2924797) and rabbit anti-inducible nitric oxide synthase (iNOS) antibody (1:50, Invitrogen, Cat# PA1-036, RRID: AB_325773) or mouse anti-Iba1 antibody and rabbit anti-Arginase 1 (Arg1) antibody (1:200, Invitrogen, Cat# PA5-85267, RRID: AB_2792410) overnight at 4°C, then Alexa Fluor 488 goat anti-rabbit antibody (1:200, Invitrogen, Cat# A-11034, RRID: AB_2576217) and Alexa Fluor 568 goat anti-mouse antibody (1:200, Invitrogen, Cat# A-11004, RRID: AB_2534072) were added at room temperature, and the tissue sections were incubated in a dark box. After incubation for 60 min, the slides were mounted with antifade reagent containing DAPI. An OlyVIA VS200 microscope was used to view and image the sections. The proportion of different types of cells was determined using ImageJ software.

Liang Z., Yang Z., Xie H., Rao J., Xu X., Lin Y., Wang C, & Chen C. (2024). Small extracellular vesicles from hypoxia-preconditioned bone marrow mesenchymal stem cells attenuate spinal cord injury via miR-146a-5p-mediated regulation of macrophage polarization. Neural Regeneration Research, 19(10), 2259-2269.

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