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Alexa fluor 555 conjugated goat anti rabbit

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Alexa Fluor 555-conjugated goat anti-rabbit is a secondary antibody used for detection and visualization in immunoassays and immunofluorescence techniques. The antibody is conjugated with the Alexa Fluor 555 fluorescent dye, which excites at 555 nm and emits at 565 nm.

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Lab products found in correlation

Alexa fluor 488 conjugated goat anti mouse, by Thermo Fisher Scientific (3 mentions) Triton x 100, by Merck Group (2 mentions) H3k4me3 millipore 07 473, by Merck Group (2 mentions) H3k9me2 millipore 07 441, by Merck Group (2 mentions) H3k27me3 millipore 07 449, by Merck Group (2 mentions) H3k27me1 millipore 07 448, by Merck Group (2 mentions) H3k36me3 9050, by Abcam (2 mentions) Vectashield mounting medium with dapi, by Vector Laboratories (2 mentions) Bx51 fluorescence microscope, by Olympus (2 mentions) Alexa fluor 488 conjugated goat anti rabbit, by Thermo Fisher Scientific (2 mentions) Lsm 7, by Zeiss (1 mentions) Zen software, by Zeiss (1 mentions) Rabbit anti neutrophil elastase antibody, by Abcam (1 mentions) Axiovert 200m microscope, by Zeiss (1 mentions) Anti ki67, by ABclonal (1 mentions) Triton x 100, by Beyotime (1 mentions) Paraformaldehyde (pfa), by Sinopharm (1 mentions) Bovine serum albumin (bsa), by Sangon (1 mentions) Sulfo nhs lc biotin, by Thermo Fisher Scientific (1 mentions) Rabbit anti myc, by Merck Group (1 mentions)

Close spelling variants of this product

Alexa Fluor 555-conjugated goat anti-rabbit antibody (10 citations) Alexa Fluor 555-conjugated goat anti-rabbit IgG secondary antibody (7 citations) Alexa Fluor 555-conjugated goat anti-rabbit IgG (35 citations) Alexa Fluor 555-conjugated goat anti-rabbit immunoglobulin G (2 citations) Alexa Fluor 555-conjugated goat anti-rabbit IgG antibody (2 citations) Goat anti-rabbit conjugated to Alexa Fluor 555 (2 citations) Alexa Fluor 555-conjugated goat anti-rabbit IgG (H + L) (4 citations) Goat anti-rabbit antibody conjugated to Alexa Fluor 555 (2 citations) Alexa Fluor 555-conjugated goat anti-rabbit secondary antibody (9 citations) Alexa Fluor” 488 or 555 conjugated goat anti-rabbit or anti-mouse antibodies (2 citations)

18 protocols using alexa fluor 555 conjugated goat anti rabbit

1

Ginseng Compounds Modulate FoxO3a Localization

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MNT1 cells were treated with each reagent [ginseng root extract (GR), ginseng berry extract (GB), and syringaresinol (SYR)] for 4 d. The cells were washed with PBS, fixed in 4% paraformaldehyde for 30 min, and incubated in 0.1% Triton X-100 (Sigma Aldrich, St. Louis, USA) for 10 min. Then, the cells were incubated with antiFoxO3a (1:200) antibody diluted in Hank's solution (0.44mM KH2PO4, 5.37mM KCl, 0.34mM Na2HPO4, 136.89mM NaCl, and 5.55mM D-glucose) overnight at 4°C. Secondary antibodies (Alexa Fluor 555-conjugated goat antirabbit, Invitrogen, Carlsbad, CA) were added for 1 h at room temperature. After washing process, the coverslips were mounted onto glass slides and visualized with a confocal laser scanning microscope (LSM7, Carl Zeiss, NY, USA; with an excitation wavelength at 555 nm and an emission wavelength at 565 nm with DAPI fluorescence). Cells with FoxO3a localization were counted and represented by the means of a confocal laser scanning microscope with the ZEN software (Carl Zeiss).
Kim J., Cho S.Y., Kim S.H., Cho D., Kim S., Park C.W., Shimizu T., Cho J.Y., Seo D.B, & Shin S.S. (2016). Effects of Korean ginseng berry on skin antipigmentation and antiaging via FoxO3a activation. Journal of Ginseng Research, 41(3), 277-283.
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2

Quantification of Mycobacterial Uptake by Neutrophils

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FITC-labeled M. avium (FITC-Mav) was prepared and bacterial localization by confocal microscopy was performed as previously described (50 (link)). FITC-Mav were sonicated, washed, and resuspended into 0.5 mL of PBS. FITC-Mav was combined with neutrophils for 1 h at a multiplicity of infection of approximately 5:1. The cells were cytocentrifuged to microscope slides and then air-dried. The slides were processed with rabbit anti-neutrophil elastase antibody (Abcam, Boston, MA), 4′,6-diamidino-2-phenylindole (DAPI), and Alexa Fluor 555-conjugated goat anti-rabbit (Invitrogen, Waltham, MA) and imaged using a Zeiss Axiovert 200M microscope. Images were quantified by calculating percent FITC(+) cells per field.
Lenhart-Pendergrass P.M., Malcolm K.C., Wheeler E., Rysavy N.M., Poch K., Caceres S., Calhoun K.M., Martiniano S.L, & Nick J.A. (2023). Deficient Complement Opsonization Impairs Mycobacterium avium Killing by Neutrophils in Cystic Fibrosis. Microbiology Spectrum, 11(1), e03279-22.
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3

Immunofluorescence Assay for Cellular Localization

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SiHa and CaSki cells were grown on glass coverslips (1×105 cells per well) in 24-well plates, fixed in 4% paraformaldehyde (Sinopharm Chemical Reagent Co., Ltd.) for 15 min at room temperature and permeabilized with 0.1% Triton X-100 (Beyotime Institute of Biotechnology) for 30 min at room temperature. After blocking with 1% BSA (Sangon Biotech Co., Ltd.) for 15 min at room temperature, the coverslips were incubated with anti-ki67 (dilution, 1:200; cat no. A11390, ABclonal Biotech Co., Ltd.) or anti-p65 antibodies (dilution, 1:200; cat no. A19653, ABclonal Biotech Co., Ltd.) at 4°C overnight, followed by incubation with the Alexa Fluor™ 555-conjugated goat anti-rabbit (dilution, 1:200; cat no. A27039, Invitrogen; Thermo Fisher Scientific, Inc.) at room temperature for 1 h. Cell nuclei were stained with DAPI (Shanghai Aladdin Biochemical Technology Co., Ltd.) at room temperature for 5 min and observed under a fluorescence microscope. Total p65 and nuclear p65 levels were quantified. Relative p65 nuclear localization was expressed as the ratio of the fluorescence density of nuclear p65 staining to the fluorescence density of total p65 cellular staining.
Deng B., Li A., Zhu Y., Zhou Y., Fei J, & Miao Y. (2023). SHCBP1 contributes to the proliferation and self‑renewal of cervical cancer cells and activation of the NF‑κB signaling pathway through EIF5A. Oncology Letters, 25(6), 246.
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4

Cell Culture and Antibody Labeling

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QM5 cells were maintained in medium 199 with Earle’s salts containing 100 U/mL of penicillin and streptomycin and 10% heat-inactivated fetal bovine serum. Monoclonal mouse anti-FLAG (Sigma-Aldrich) and rabbit anti-Myc (Sigma-Aldrich) antibodies, horseradish peroxidase (HRP)-conjugated goat anti-rabbit (Santa Cruz) and goat-anti mouse (Santa Cruz) antibodies, Alexa Fluor 488-conjugated goat anti-mouse (Invitrogen) and Alexa Fluor 555-conjugated goat anti-rabbit (Invitrogen), Sulfo-NHS-LC-Biotin (Thermo Scientific), maliemide-PEG2-biotin (Thermo Scientific), neutravidin agarose resin (Thermo Scientific), and polyethylimine (PEI, Polysciences Inc.) were purchased from the indicated commercial sources.
Yang Y., Gaspard G., McMullen N, & Duncan R. (2020). Polycistronic Genome Segment Evolution and Gain and Loss of FAST Protein Function during Fusogenic Orthoreovirus Speciation. Viruses, 12(7), 702.
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5

Methylation analysis of ELF6 mutant in N. benthamiana

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Wild-type ELF6 or mutant elf6-5 full genomic coding sequences were cloned into pEG104 vector35 (link). The demethylation assay was carried out as previously described16 (link). Briefly, N. benthamiana leaves were infiltrated with A. tumefaciens EHA105 strains carrying a functional wild-type 35S::YFP::ELF6 or mutant 35S::YFP::ELF6A424V. Transfected nuclei were isolated after 48 h. Immunolabeling of fixated nuclei was performed using histone methylation-specific antibodies: H3K4me3 Millipore 07-473, 1:100; H3K9me2 Millipore 07-441, 1:200; H3K27me3 Millipore 07-449, 1:100; H3K27me2 Millipore 07-452, 1:200; H3K27me1 Millipore 07-448, 1:200; H3K36me3 Abcam 9050, 1:100. The modified histones were revealed by Alexa Fluor 555-conjugated goat anti-rabbit (Invitrogen, 1:200). Transfected cells were revealed by monitoring the YFP signal. After staining, the slides were mounted in VECTASHIELD mounting medium with DAPI (Vector Laboratory) and then photographed with an OLYMPUS BX51 fluorescence microscope. Histone methylation levels were quantified by comparing staining density of a number of transfected 35S::YFP::ELF6 nuclei versus non-transfected nuclei in the same field. Image density was determined using ImageJ software. A negative result in our assay usually corresponds to 80% or less than total wild-type histone demethylase activity.
Crevillén P., Yang H., Cui X., Greeff C., Trick M., Qiu Q., Cao X, & Dean C. (2014). Epigenetic reprogramming that prevents transgenerational inheritance of the vernalized state. Nature, 515(7528), 587-590.
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6

Immunofluorescence Staining of COX-2, Tubulin, Ki-67, and F4/80

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Immunofluorescence staining was used to detect COX-2 and tubulin in RAW264.7 cells. The anti-COX-2 antibody (CST, dilution 1:200) was incubated overnight at 4°C and exposed to Alexa Fluor-555-conjugated goat-anti-rabbit (Invitrogen, dilution 1:500) antibody for 1 h at RT. After that, the anti-tubulin antibody (Abcam, dilution 1:200) was also incubated overnight at 4°C and exposed to Alexa Fluor-488-conjugated goat-anti-rabbit (Invitrogen, dilution 1:500) antibody. In addition, immunofluorescence staining was also used to detect Ki-67 and F4/80 in the gum tissue. The anti-Ki-67 antibody (Invitrogen, dilution 1:100) and the anti-F4/80 antibody (Abcam, dilution 1:50) were used as primary antibodies. After the removal of first antibodies, they were exposed to goat anti-mouse IgG H&L (Alexa Fluor® 594, Abcam, United States, dilution 1:500) and Alexa Fluor-488-conjugated goat-anti-rabbit (Invitrogen, dilution 1:500). Nuclei were counterstained with DAPI solution. Images were analyzed using a fluorescence confocal microscope.
Chen L., Zhao T., Liu M., Chen Q., Yang Y., Zhang J., Wang S., Zhu X., Zhang H., Huang Q, & Ai K. (2022). Ultra-small molybdenum-based nanodots as an antioxidant platform for effective treatment of periodontal disease. Frontiers in Bioengineering and Biotechnology, 10, 1042010.
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7

Immunohistochemical Detection of VGLUT1 and VGLUT2

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To determine expression of VGLUT1 and VGLUT2, brain sections were incubated in blocking solution (PBS with 1% normal goat serum [Jackson ImmunoResearch laboratories, Inc.] and 0.1% Triton-X) for 30 minutes, to prevent non-specific binding of the secondary antibody. Subsequently, sections were incubated for 24h with a primary antibody against VGLUT1 (polyclonal rabbit anti-VGLUT1, diluted 1:1000 in blocking solution; Synaptic Systems, Cat. #135 303) or VGLUT2 (polyclonal rabbit anti-VGLUT2, diluted 1:1000 in blocking solution; Synaptic Systems, Cat. #135 403). The next day, primary antibody binding was visualized by a 2h-incubation with a fluorescent secondary antibody (Alexa Fluor 555-conjugated goat anti-rabbit, diluted 1:500 in blocking solution; Invitrogen). Sections were cover slipped with Fluoromount-G (eBioscience Inc.). To ensure specificity of the detection method, negative controls for the secondary antibody were obtained, in which sections were not treated with the primary antibody. Specificity of the primary antibodies in cochlear nucleus and cerebellar tissue has been shown previously in our lab (Zhou et al., 2007 (link)). Positive controls for VGLUT-immunolabeling were performed in the cerebellar cortex (Takamori et al., 2001 (link); Kaneko et al., 2002 (link); Hioki et al., 2003 (link)). All procedures were performed at room temperature.
Heeringa A.N., Wu C., Chung C., West M., Martel D., Liberman L., Liberman M.C, & Shore S.E. (2018). Glutamatergic Projections to the Cochlear Nucleus are Redistributed in Tinnitus. Neuroscience, 391, 91-103.
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8

Immunofluorescence Staining of Neural Crest Cells

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Ten days after isolation and culture, the cells were passaged and plated on collagen-coated cover slips overnight, washed in PBS and fixed in 4% paraformaldehyde for 10 min. The fixed cells were washed in PBS and incubated in blocking buffer containing 10% goat serum (Sigma-Aldrich, St. Louis, MO, USA) and 0.3% Triton X-100 (Fluka, St. Louis, MO, USA) at room temperature for 30 min. Cells were then incubated at 4 °C overnight with one of the following primary antibodies: anti-SOX10 (1:200, rabbit polyclonal IgG; Abcam, Cambridge, MA, USA), anti-Snail 1 (1:100, rabbit polyclonal IgG; Santa Cruz, Biotechnology, Inc., Santa Cruz, CA, USA), or anti-p75 (1:400, rabbit polyclonal IgG; Abcam). The next day, the cells were rinsed three times to remove the unbound primary antibodies and then incubated at room temperature for 2 h with secondary antibody (Alexa Fluor 555-conjugated goat anti-rabbit,1:400; Invitrogen). The cell nuclei were counterstained with 4′,6-diamidino-2-phenylindole (DAPI, 1 μg/mL; Invitrogen) in PBS, in the dark and at room temperature for 1 min. After washing, the stained cover slips were removed, mounted on a slide with mounting media, and visualized using a fluorescence microscope (LeicaDM2500; Leica Microsystems GmbH, Wetzlar, Germany) equipped with an EMCCD camera (LucaEM R DL-604M; Andor Technology, Belfast, UK).
Peng C.K., Wu S.Y., Tang S.E., Li M.H., Lin S.S., Chu S.J, & Huang K.L. (2017). Protective Effects of Neural Crest-Derived Stem Cell-Conditioned Media against Ischemia-Reperfusion-Induced Lung Injury in Rats. Inflammation, 40(5), 1532-1542.
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9

Antibody Panel for Phosphorylation Analysis

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Primary antibodies including anti-phospho-mTOR (Ser2448), anti-mTOR, anti-phospho-4EBP1 (Ser65), anti-4EBP1, anti-phospho-Akt (Ser473), anti-phospho-Akt (Thr308), anti-Akt, anti-phospho-rpS6 (Ser240/244), anti-phospho-rpS6 (Ser235/236), anti-rpS6, anti-p-p53 (Ser15), p53, anti-phospho-IGF-1Rβ (T1135/36/1150/51), IGF-1Rβ and anti-MYC antibodies were purchased from Cell Signaling Technology (Danfoss, MA, USA). The anti-Rabbit IgG antibody was purchased from Jackson ImmunoResearch Laboratories (West Grove, PA, USA). Anti-S6K1 antibody was purchased from Abcam (Cambridge, UK). Anti-β-actin, anti-FLAG (M2) and anti-p62/SQSTM1 antibodies were purchased from Sigma (Saint Louis, MO, USA), anti-LC3 antibody was purchased from Medical & Biological Laboratories (Nagoya, Aichi, Japan). Secondary antibodies used in western blotting including IRDye 800CW Donkey anti-Mouse and anti-Rabbit IgG were purchased from Li-COR (Lincoln, NE, USA). Secondary antibodies used in immunofluorescence including AlexaFluor-555-conjugated goat anti-mouse and AlexaFluor-555-conjugated goat anti-rabbit antibodies were purchased from Invitrogen (Burlingame, CA, USA).
Lu T., Zhu Z., Wu J., She H., Han R., Xu H, & Qin Z.H. (2019). DRAM1 regulates autophagy and cell proliferation via inhibition of the phosphoinositide 3-kinase-Akt-mTOR-ribosomal protein S6 pathway. Cell Communication and Signaling : CCS, 17, 28.
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10

Immunofluorescence Staining of 3D Cell Cultures

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MCF10A-5E and MCF10DCIS.com 3D cultures were embedded at day 10 of morphogenesis, and 5-μm sections were cut and mounted on Superfrost Plus slides (Fisher). For clinical samples, paraffin tissue sections were dewaxed and antigens retrieved on a PT Link (Dako) with low-pH EnVision FLEX Target Retrieval Solution (Dako) for 20 min at 97°C. Immunofluorescence on cryosections and antigen-retrieved slides was performed as previously described (20 (link)) with the following primary antibodies: NRF2 (Santa Cruz #sc-13032, 1:100), phospho-Rb (Cell Signaling #8516, 1:1600), HIF-1α (Cell Signaling #79233, 1:800), p53 (Santa Cruz #sc-126, 1:200). Slides were incubated the next day for 1 hour in the following secondary antibodies: Alexa Fluor 555-conjugated goat anti-rabbit (1:200; Invitrogen), Alexa Fluor 647-conjugated goat anti-mouse (1:200; Invitrogen).
Pereira E.J., Burns J.S., Lee C.Y., Marohl T., Calderon D., Wang L., Atkins K.A., Wang C.C, & Janes K.A. (2020). Sporadic activation of an oxidative stress-dependent NRF2–p53 signaling network in breast epithelial spheroids and premalignancies. Science signaling, 13(627), eaba4200.
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