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Goat anti rabbit alexa568

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Goat anti-rabbit Alexa568 is a secondary antibody conjugated with the Alexa Fluor 568 fluorescent dye. It is designed to detect and visualize rabbit primary antibodies in immunological applications.

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Lab products found in correlation

Goat anti mouse alexa 488, by Thermo Fisher Scientific (9 mentions) Goat anti chicken alexa 488, by Thermo Fisher Scientific (4 mentions) Goat anti rabbit alexa 488, by Thermo Fisher Scientific (3 mentions) Goat anti mouse alexa 568, by Thermo Fisher Scientific (3 mentions) Rabbit anti gfp, by Thermo Fisher Scientific (2 mentions) Anti mouse alexa 488, by Thermo Fisher Scientific (2 mentions) Axioplan 2 microscope, by Zeiss (2 mentions) α h3p, by Merck Group (2 mentions) Goat anti mouse alexa 647, by Thermo Fisher Scientific (2 mentions) Ab40673, by Abcam (1 mentions) Chicken anti tyrosine hydroxylase, by Aves Labs (1 mentions) Donkey anti rabbit hrp, by Thermo Fisher Scientific (1 mentions) Donkey anti chicken alexa 488, by Jackson ImmunoResearch (1 mentions) Goat anti mouse alexa633, by Thermo Fisher Scientific (1 mentions) Fluoview fv1000, by Olympus (1 mentions) Mouse anti gfp, by Merck Group (1 mentions) Normal goat serum (ngs), by Merck Group (1 mentions) Fv3000, by Olympus (1 mentions) Rabbit anti dsred, by Takara Bio (1 mentions) Fluoroshield mounting medium, by Abcam (1 mentions)

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Alexa568 anti-rabbit (2 citations)

28 protocols using goat anti rabbit alexa568

1

Antibodies and Reagents for Dopamine Analysis

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Primary antibodies used in this study include Rabbit anti-Rho (abcam Ab40673), Rabbit Anti Rho-S188P (abcam Ab41435), Rabbit anti-DAT (Amara Lab generated), Rabbit anti-EAAT3 (Alpha diagnostics EAAC11-A), Chicken anti-Tyrosine Hydroxylase (Aves Labs, Inc. TYH), and Rabbit anti-G13 (abcam Ab128900). We used the following secondary antibodies: Donkey anti-chicken Alexa488 (Jackson Immuno Research Labs 120990), Goat anti-rabbit Alexa-568 (Thermo Fisher Scientific A-11036), and Donkey anti-rabbit HRP (Thermo Scientific 31458). Our custom peptides were synthesized by LifeTein. The (+)-Amphetamine hemisulfate [(αS)-α-Methylbenzeneethanamine sulfate] provided by the National Institute on Drug Abuse Drug Supply Program Division of Therapeutics and Medical Consequences. All other drugs used in this study were from Sigma-Aldrich or Tocris.
Underhill S.M., Hullihen P.D., Chen J., Fenollar-Ferrer C., Rizzo M.A., Ingram S.L, & Amara S.G. (2019). Amphetamines signal through intracellular TAAR1 receptors coupled to Gα13 and GαS in discrete subcellular domains. Molecular Psychiatry, 26(4), 1208-1223.
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2

Immunostaining and Brain Registration Protocol

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Brains from 7-to-10-day-old adult files were dissected and stained as previously described(Watanabe et al., 2017 (link)). The primary antibody mixture consisted of 1:1000 rabbit anti-GFP (Thermo Fisher Scientific, Cat#A11122), 1:1000 chicken anti-GFP (Aves Lab, Cat#GFP-1010), 1:100 mono-clonal (for GRASP experiment, Figure S4J–R) mouse anti-GFP (Sigma-Aldrich, Cat#G6539), 1:1000 rabbit anti-DsRed (Takara Bio, Cat#632496), 1:50 mouse anti-Brochpilot nc82 (Developmental Studies Hybridoma Bank), and 10% normal goat serum (Sigma-Aldrich) in PBST. Secondary antibodies used were 1:1000 goat anti-rabbit-Alexa488 (Thermo Fisher Scientific, Cat#A11008), 1:1000 goat anti-chicken-Alexa488 (Thermo Fisher Scientific, Cat#A11039), 1:1000 goat anti-mouse-Alexa488 (Thermo Fisher Scientific, Cat#A11001), 1:1000 goat anti-rabbit-Alexa568 (Thermo Fisher Scientific, Cat#A11011), and 1:1000 goat anti-mouse-Alexa633 (Thermo Fisher Scientific, Cat#A21050).
Confocal stacks were obtained with Fluoview FV1000 or FV3000 (Olympus). Fiji (Schindelin et al., 2012 (link); Schneider et al., 2012 (link)) and Fluorender (Wan et al., 2009 (link)) software was used to create z stack images. For brain registration (Figure S4G–I), the two images shown in Figure S4B and D are registered to T1 template brain (Yu et al., 2010 (link)) using CMTK registration tools (Jefferis et al., 2007 (link)).
Jung Y., Kennedy A., Chiu H., Mohammad F., Claridge-Chang A, & Anderson D.J. (2019). NEURONS THAT FUNCTION WITHIN AN INTEGRATOR TO PROMOTE A PERSISTENT BEHAVIORAL STATE IN DROSOPHILA. Neuron, 105(2), 322-333.e5.
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3

Immunofluorescent Staining for Epigenetic and Proliferation Markers

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Tissues, sectioning, and dehydration were completed in the same manner as Immunohistochemistry samples. Antigen retrieval was also performed as described above, with the exception of staining for OLFM4 and GFP, in which slides underwent antigen retrieval in 10 mM Tris-EDTA pH 9.0 (10 mM Tris, 10 mM EDTA). Slides were then washed in 1× PBS with 0.05% Tween-20, and blocked in 10% goat serum in this washing solution for 1 h at room temperature. Slides were washed in washing solution, and incubated in primary antibodies diluted in blocking solution at 4 °C overnight in a humidified chamber. Primary antibodies were used as follows: rabbit H3K27me3 (1:750; EMD Millipore, 07-449), mouse PCNA (1:300; Santa Cruz Biotechnology, sc-56), rabbit OLFM4 (1:100; Cell Signaling Technology, D6Y5A), mouse GFP (1:300; Santa Cruz Biotechnology, 9996). Goat anti-mouse Alexa 488 (Thermo Fisher Scientific, A11029), goat anti-mouse Alexa 568 (Thermo Fisher Scientific, A11031), and goat anti-rabbit Alexa 568 (Thermo Fisher Scientific, A11036) were used as secondary antibodies at 1:500 in washing buffer with nuclear Hoescht 33342 stain (1:1000; Thermo Fisher Scientific, 62249) for 1 h at room temperature. Slides were mounted with Fluoroshield Mounting Medium (Abcam, 104135) and sealed with clear nail polish for preservation.
Smith R.J., Liang M., Loe A.K., Yung T., Kim J.E., Hudson M., Wilson M.D, & Kim T.H. (2023). Epigenetic control of cellular crosstalk defines gastrointestinal organ fate and function. Nature Communications, 14, 497.
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4

Immunofluorescence Staining of LC3B

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Cells were washed 1 × with PBS, fixed for 15 min with 4% PFA at room temperature or with 100% cold methanol on ice (LC3-staining). For immunostaining, cells were washed 3 ×, permeabilized at room temperature with 0.1% TX-100 for 10 min and washed again 1 ×. Non-specific binding was blocked using 3% goat serum (Gibco) in PBS (blocking solution) for 30 min and the cells were then incubated with 1:200 diluted rabbit anti LC3B (Sigma, L7543) in blocking solution for 2 h. After washing 2 ×, cells were incubated with 1:400 diluted goat anti rabbit Alexa 568 (ThermoFisher Scientific, A-11011) for 1 h. After washing 2 ×, cells were stained with 1:10,000 diluted Hoechst 33342 (ThermoFisher Scientific, H1399) for 10 min and washed once.
Heintze J., Costa J.R., Weber M, & Ketteler R. (2016). Ribose 5-phosphate isomerase inhibits LC3 processing and basal autophagy. Cellular Signalling, 28(9), 1380-1388.
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5

Antibody Panel for Cell Signaling

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The following antibodies were used: Phospho-S473-KAP1 (BioLegend Cat# 654102, RRID: AB_2561782), phospho-S516-CHK2 (Cell Signaling Technology Cat# 2669, RRID: AB_330146), CHK2 (Abcam Cat# ab109413, RRID: AB_10863751), KAP1 (GeneTex Cat# GTX102227, RRID: AB_2037323), PCNA (Santa Cruz Biotechnology Cat# sc-56, RRID:AB_628110), goat anti-mouse (Thermo Fisher Scientific Cat# A-11004, RRID: AB_2534072), and goat anti-rabbit Alexa568 (Thermo Fisher Scientific Cat# A-11011, RRID: AB_143157).
Stolarova L., Kleiblova P., Zemankova P., Stastna B., Janatova M., Soukupova J., Achatz M.I., Ambrosone C., Apostolou P., Arun B.K., Auer P., Barnard M., Bertelsen B., Blok M.J., Boddicker N., Brunet J., Burnside E.S., Calvello M., Campbell I., Chan S.H., Chen F., Chiang J.B., Coppa A., Cortesi L., Crujeiras-González A., De Leeneer K., De Putter R., DePersia A., Devereux L., Domchek S., Efremidis A., Engel C., Ernst C., Evans D.G., Feliubadaló L., Fostira F., Fuentes-Ríos O., Gómez-García E.B., González S., Haiman C., Hansen T.V., Hauke J., Hodge J., Hu C., Huang H., Ishak N.D., Iwasaki Y., Konstantopoulou I., Kraft P., Lacey J., Lázaro C., Li N., Lim W.K., Lindstrom S., Lori A., Martinez E., Martins A., Matsuda K., Matullo G., McInerny S., Michailidou K., Montagna M., Monteiro A.N., Mori L., Nathanson K., Neuhausen S.L., Nevanlinna H., Olson J.E., Palmer J., Pasini B., Patel A., Piane M., Poppe B., Radice P., Renieri A., Resta N., Richardson M.E., Rosseel T., Ruddy K.J., Santamariña M., Dos Santos E.S., Teras L., Toland A.E., Trentham-Dietz A., Vachon C.M., Volk A.E., Weber-Lassalle N., Weitzel J.N., Wiesmuller L., Winham S., Yadav S., Yannoukakos D., Yao S., Zampiga V., Zethoven M., Zhang Z.W., Zima T., Spurdle A.B., Vega A., Rossing M., Del Valle J., De Nicolo A., Hahnen E., Claes K.B., Ngeow J., Momozawa Y., James P.A., Couch F.J., Macurek L, & Kleibl Z. (2023). ENIGMA CHEK2gether Project: A Comprehensive Study Identifies Functionally Impaired CHEK2 Germline Missense Variants Associated with Increased Breast Cancer Risk. Clinical Cancer Research, 29(16), 3037-3050.
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6

Immunofluorescence Imaging of H3K27me3 and Embryonic Myosin

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Immunofluorescence was performed as described previously (Gnocchi et al., 2009) with modifications, using rabbit anti-H3K27me3 (Milipore) and mouse anti-MyHC-emb (eMyHC, Developmental Hybridoma bank) antibodies. Secondary antibodies were goat anti-rabbit-Alexa568 and anti-mouse-Alexa568 (ThermoFisher).
Perovanović J., Wu Y., Abewe H., Shen Z., Hughes E.P., Gertz J., Chandrasekharan M.B., & Tantin D. (2020). Oct1 cooperates with Smad transcription factors to promote mesodermal lineage specification.
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7

Immunofluorescence Staining of HUVECs

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Cells were cultured on nanopillars or 8-well chamber slides (Sigma-Aldrich), cells were fixed with 4% PFA (P6148, Sigma-Aldrich) for 15 min at room temperature (RT). The cell membrane was then permeabilized with 0.1% Triton X-100 (X100, Sigma-Aldrich) in PBS (10 min) followed by 1% BSA/PBS (85040C, Sigma-Aldrich) blocking step (30 min, RT).
HUVECs were stained with with mouse anti-NCL (1:100, Santa Cruz), rabbit anti-HK2 (1:100, proteintech), rabbit anti-PFKFB3 (1:100, abcam, rabbit anti-CPT1a (1:500, abcam), and rabbit andti-TIGAR (1:200, Sigma) in 1% BSA/PBS at 4°C overnight. Cells were the incubated with secondary antibodies goat anti-rabbit Alexa 568 and goat anti-mouse Alexa 488 (1:500, Thermo Fisher Scientific), or TRITC-labeled phalloidin (1:100 Sigma) in 1% BSA/PBS for 1.5 hours at RT. Nuclei were counterstained with DAPI staining using (1:20,000, Thermo Fisher Scientific) in PBS for 5 min.
Schwab M., Trizio I.d., Shiu J., Ghobrial M., Sürücü O., Girolamo F., Errede M., Yılmaz M., Haybaeck J., Moiraghi A., Monnier P.P., Lawler S.E., Greenfield J.P., Radovanovic I., Frei K., Schlapbach R., Vogel V., Virgintino D., Bock K.D., & Wälchli T. (2020). Nucleolin is expressed in human fetal brain development and reactivated in human glial brain tumors regulating angiogenesis and vascular metabolism.
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8

Immunofluorescence Imaging of Gonadal Proteins

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Gonads were dissected from 1-day adults of him-17::3xHA worms in 1X sperm salts (50 mM PIPES pH 7.0, 25 mM KCl, 1 mM MgSO4, 45 mM NaCl, and 2 mM CaCl2) with 1 mM levamisole and fixed in 1% paraformaldehyde diluted in 1X sperm salts for 5 min in a humid chamber. Slides were then frozen on a metal block on dry ice for at least 10 min prior to flicking off the coverslip and immersing in 100% ethanol at −20° C for 2 min. Slides were then washed in PBSTB [1XPBS with 0.1% Tween and 0.1% bovine serum albumin (BSA)], and incubated overnight at 4°C with primary antibody: mouse anti-HA (Cell Signaling), guinea pig anti-XND-1(Wagner et al., 2010 (link)), and rabbit anti-HTP-3 (Das et al., 2022 (link)) (all diluted 1:2000 in PBST). The next day, slides were washed 3X in PBSTB for 10 min and incubated with secondary antibodies: anti-mouse Alexa 488 (Molecular Probes), anti-guinea pig 633, and goat anti-rabbit Alexa 568 (Invitrogen, all diluted 1:2000 in PBSTB) for 2 h at room temperature in the dark. Slides were then washed 2 X 10 min in PBSTB, mounted in Prolong Gold without DAPI (Invitrogen) and put in the dark to cure overnight before imaging. The images were taken using a Stellaris8 TauSTED equipped with three depletion lines.
Raices M., Balmir F., Silva N., Li W., Grundy M.K., Hoffman D.K., Altendorfer E., Camacho C.J., Bernstein K.A., Colaiácovo M.P, & Yanowitz J. (2024). Genetic and physical interactions reveal overlapping and distinct contributions to meiotic double-strand break formation in C. elegans. bioRxiv.
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9

Visualization of Serum-Complexed NBD-Tocopherol

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Serum-complexed NBD-tocopherol (34 (link)) was loaded into primary cultures grown on 50 μg/ml poly-l-lysine–coated cover slips overnight. After washing, the cells were fixed in 3.7% paraformaldehyde, permeabilized in 5% goat serum/0.3% Triton X-100, followed by incubation with anti-TTP, anti-β-tubulin III (Sigma), or anti-GFAP (Becton Dickenson) antibodies. Goat anti-rabbit Alexa 568 (Invitrogen) was used to visualize TTP, whereas goat antimouse Alexa 488 (Invitrogen) was the secondary antibody for β-tubulin III and GFAP. 4′,6-Diamidino-2-phenylindole staining was used to visualize nuclei. After mounting in SlowFade Gold antifade reagent (Invitrogen), slides were imaged on an inverted (Leica DM4100B) or confocal (Zeiss LSM 510) microscope. Cerebellar slice cultures were immunostained according to the previous published technique (114 (link)) using the same antibodies.
Ulatowski L., Ghelfi M., West R., Atkinson J., Finno C.J, & Manor D. (2022). The tocopherol transfer protein mediates vitamin E trafficking between cerebellar astrocytes and neurons. The Journal of Biological Chemistry, 298(3), 101712.
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10

Microfibril Incorporation Assay in Fibroblast-HEK293T Co-cultures

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Microfibril incorporation assays were carried out as described previously [16 (link), 26 (link)] with some modifications. Briefly, HEK293T cells were transfected in 6-well plates overnight then trypsinised and counted. Co-cultures on 13 mm diameter glass coverslips were established using 2 x 105 FS2 dermal fibroblasts [14 (link)] and 2 x 105 transfected HEK293T cells per well of 24 well plates. Cells were cultured in complete DMEM for five days then fixed with 4% (w/v) paraformaldehyde in phosphate-buffered saline and stained using a rabbit polyclonal antibody raised against the FBN1 proline-rich region [32 (link)] and chicken polyclonal anti-GFP (Abcam; ab13970) without permeabilisation. Goat anti-chicken Alexa488 and goat anti-rabbit Alexa568 (Invitrogen) were used for detection. Images were collected using a Zeiss Axioplan 2 microscope with AxioVision Rel. 4.8 software.
Jensen S.A., Atwa O, & Handford P.A. (2021). Assembly assay identifies a critical region of human fibrillin-1 required for 10–12 nm diameter microfibril biogenesis. PLoS ONE, 16(3), e0248532.
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