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Anti rabbit igg alexa fluor 568

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The Anti-rabbit IgG Alexa Fluor 568 is a secondary antibody conjugated with the Alexa Fluor 568 fluorescent dye. It is designed to detect and visualize rabbit primary antibodies in various immunoassay techniques.

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Lab products found in correlation

Alexa fluor 488 anti mouse igg, by Thermo Fisher Scientific (20 mentions) 4 6 diamidino 2 phenylindole (dapi), by Thermo Fisher Scientific (8 mentions) Alexa fluor 568 anti mouse igg, by Thermo Fisher Scientific (6 mentions) Alexa fluor 488 anti rabbit igg, by Thermo Fisher Scientific (6 mentions) Alexa fluor 488 phalloidin, by Thermo Fisher Scientific (5 mentions) 4 6 diamidino 2 phenylindole (dapi), by Dojindo Laboratories (4 mentions) Saponin, by Merck Group (3 mentions) Triton x 100, by Merck Group (3 mentions) Alexa fluor 488, by Thermo Fisher Scientific (3 mentions) Alexa fluor 488 anti rat igg, by Thermo Fisher Scientific (3 mentions) Mitotracker red, by Thermo Fisher Scientific (3 mentions) Lsm 710 confocal microscope, by Zeiss (3 mentions) Normal goat serum (ngs), by Agilent Technologies (2 mentions) Ag 25b 0006 c100, by Adipogen (2 mentions) Fam flica active caspase 1, by ImmunoChemistry Technologies (2 mentions) C2 confocal microscope, by Nikon (2 mentions) Alexa fluor 488 anti goat igg, by Thermo Fisher Scientific (2 mentions) Mouse anti α tubulin, by Merck Group (2 mentions) Rabbit anti β actin, by GeneTex (2 mentions) Rabbit anti paxillin, by GeneTex (2 mentions)

43 protocols using anti rabbit igg alexa fluor 568

1

Ciona Embryo Immunofluorescence Staining

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Ciona embryos were fixed with stationary liquid [19 (link)] (100 mM HEPES, pH = 6.9; 100 mM EGTA, pH = 7.0; 10 mM MgSO4; 2% formaldehyde; 0.1% glutaraldehyde; 300 mM dextrose; and 0.2% Triton X-100) for 40 min at room temperature. After three washes with PBS, the embryos were incubated in PBST (PBS with 0.1% Triton X-100) for 30 min to achieve permeabilization. Then, the embryos were treated with 0.1% sodium borohydride in PBS for 20 min at room temperature. Next, a 1:250 dilution of phospho-myosin light chain 2 (Ser19) antibody (#3671; Cell Signaling) was added and incubated at room temperature for 24 h. After three washes with PBS, a 1:200 dilution of Alexa Fluor 568 anti-Rabbit IgG (A11011; Invitrogen) was added and incubated for 48 h at room temperature. As needed for cell boundary visualizing, a 1:200 dilution of Alexa Fluor 488 Phalloidin (A12379; Invitrogen) was added. Finally, after three washes, the embryos were coated with mounting medium with DAPI and mounted for further observation.
Qiao J., Peng H, & Dong B. (2023). Development and Application of an Optogenetic Manipulation System to Suppress Actomyosin Activity in Ciona Epidermis. International Journal of Molecular Sciences, 24(6), 5707.
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2

Immunostaining of Blastocyst Embryos

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Only blastocyst stage embryos at time of collection were processed for immunostaining as previously described (Ross et al., 2008 (link)). Embryos were washed with phosphate buffered saline (PBS; GIBCO) containing 1 mg/mL PVA (PBS-PVA) three times and then fixed in 4% paraformaldehyde containing 1 ml/mL PVA for 15 min at room temperature. After washing three times with PBS-PBA, blastocysts were permeabilized with PBS-PVA containing 1% Triton X-100 for 30 min, washed in PBS-PVA containing 0.1% Triton X-100 (washing buffer; WB), and blocked in PBS-PVA supplemented with 10% normal donkey serum. Embryos were incubated with primary antibodies (rabbit anti-SOX2 antibody, BioGenex Cat# NU579-UC and mouse anti-human nuclei antibody, Millipore Cat# MAB1281) overnight at 4°C. After repeated washes in WB, the embryos were incubated with secondary antibodies (Alexa Fluor 568 anti-rabbit IgG and Alexa Fluor 488 anti-mouse IgG, Invitrogen) at room temperature for 1 hr. Then, blastocysts were counter-stained with 10 μg/ml Hoechst 33342 at room temperature for 20 min. Stained blastocysts were mounted on a glass slide containing ProLong Gold antifade solution (Invitrogen), covered by a coverslip, and imaged using an inverted fluorescence microscope.
Wu J., Platero-Luengo A., Sakurai M., Sugawara A., Gil M.A., Yamauchi T., Suzuki K., Bogliotti Y.S., Cuello C., Valencia M.M., Okumura D., Luo J., Vilariño M., Parrilla I., Soto D.A., Martinez C.A., Hishida T., Sánchez-Bautista S., Martinez-Martinez M.L., Wang H., Nohalez A., Aizawa E., Martinez-Redondo P., Ocampo A., Reddy P., Roca J., Maga E.A., Esteban C.R., Berggren W.T., Delicado E.N., Lajara J., Guillen I., Guillen P., Campistol J.M., Martinez E.A., Ross P.J, & Belmonte J.C. (2017). Interspecies Chimerism with Mammalian Pluripotent Stem Cells. Cell, 168(3), 473-486.e15.
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3

Immunofluorescent Staining of CXCR5+ CD4+ Cells

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Freshly isolated PP and MLN samples were fixed in 10% neutral buffered formalin and embedded in paraffin. These samples were subjected to Hematoxylin and Eosin staining along with immunofluorescent staining. Four μm sections were incubated in 5% skim milk and 1% BSA containing PBS for 1 hour and stained with rabbit anti- mouse CXCR5 polyclonal antibody (1:100) (Bioss) overnight at 4°C followed by Alexa fluor 568 anti-rabbit IgG (1:200) (Invitrogen) for 90 min at room temperature. Sections were then re-incubated with 5% skim milk for one hour followed by staining with anti-mouse CD4 (1:200) (Coulter). Goat Anti-Mouse IgGAM - FITC (1:200) (Zymed) was used as the secondary antibody. Sections were counterstained with 4′,6-Diamidino-2-Phenylindole, Dihydrochloride (DAPI) and staining specificity was confirmed by isotype-matched antibodies. Stained slides were examined using ZEISS Observer. Z1 fluorescent microscope and analyzed using Zen software. CD4+CXCR5+ cells were separately counted inside and in the surroundings of a GC under 20× magnification. Total of 5 PPs from 3 WT and TrifLPS2 mice each from baseline, and 7 PPs from 5 WT and TrifLPS2 mice each from day 9 post infection group were analyzed.
Kanagavelu S., Flores C., Hagiwara S., Ruiz J., Hyun J., Cho E.E., Sun F., Romero L., Shih D.Q, & Fukata M. (2016). TIR-Domain-Containing Adapter-Inducing Interferon-β (TRIF) Regulates CXCR5+ T helper Cells in the Intestine. Journal of clinical & cellular immunology, 7(5), 458.
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4

Immunofluorescent Labeling of Neuron-Specific Proteins

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Serial brain sections were incubated overnight at room temperature with a mixture of the antibodies anti-p-Cofilin (1:300; rabbit, ref. 47281, Abcam; Cambridge, UK) and the neuronal marker anti-NeuN (1:1000 mouse anti-NeuN; ref. VMA 377, Abcys) diluted in PBS-T-azide. Following washes, sections were incubated 2 h in Alexa Fluor® 568 anti-rabbit IgG and Alexa Fluor® 488 anti-mouse IgG (Invitrogen; California, USA) diluted 1:1000 in PBS-T. Sections were washed and mounted in Vectashield medium.
Thibault K., Rivière S., Lenkei Z., Férézou I, & Pezet S. (2016). Orofacial Neuropathic Pain Leads to a Hyporesponsive Barrel Cortex with Enhanced Structural Synaptic Plasticity. PLoS ONE, 11(8), e0160786.
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5

Immunostaining Characterization of Cellular Markers

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Immunostaining was performed as previously described.49 (link) Primary antibodies used were:
anti-mouse keratin 6 (Covance, Emeryville, California, cat#PRB-169P), anti-human
p63 (Santa Cruz, Dallas, Texas, cat#sc8431), and anti-human E-cadherin (BD
Transduction Laboratories, San Jose, California, cat#610182). Secondary
antibodies were: Alexa Fluor 488 anti-mouse IgG (Invitrogen, Carlsbad,
California cat #A-11017), Alexa Fluor 568 anti-rabbit IgG (Invitrogen, Carlsbad,
California, cat#A-11004). Nuclear DNA was labeled with DAPI
(4′,6-diamidino-2-phenylindole). Mountant was ProLong Diamond containing
DAPI (Life Technologies, Eugene, Oregon). Immunofluorescent images were acquired
with a Zeiss 700 Confocal, processed using NIH Image J and presented as maximum
projection of two to four confocal slices.
Paul B.J., Palmer K.J., Rhea L., Carlson M., Sharp J.C., Pratt C.H., Murray S.A, & Dunnwald M. (2021). The Mafb cleft-associated variant H131Q is not required for palatogenesis in the mouse. Developmental dynamics : an official publication of the American Association of Anatomists, 250(10), 1463-1476.
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6

Immunohistochemical Analysis of MAPK

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The immunohistochemistry procedure has been described previously (9) . The primary antibody was rabbit antiphospho-p44/42 MAPK (ERK1/2) polyclonal antibody (1:500, #9101, Lot #26, Cell Signaling Technology, Danvers, MA, USA), and the secondary antibody was Alexa Fluor 568 anti-rabbit IgG (1:200, A11004, Invitrogen, Paisley, UK).
Okada S., Katagiri A., Saito H., Lee J., Ohara K., Iinuma T, & Iwata K. (2019). Functional involvement of nucleus tractus solitarii neurons projecting to the parabrachial nucleus in trigeminal neuropathic pain. Journal of oral science, 61(2).
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7

Fluorescence Staining for RPTPσ Detection

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For fluorescence staining for RPTPσ, the C-terminal (His)6 tag was used to detect bound RPTPσ on the sections. Free-floating sections were first incubated with RPTPσ-AP. After washing in PBS, the sections were blocked for 1 hour with 10% goat serum (Invitrogen) in PBS containing 0.1% Triton X-100 (Sigma) followed by incubation with an anti-(His)6 antibody (Abcam) and different antibodies: rabbit anti-GFAP (DAKO), NeuN (Millipore) or biotinylated sheep anti-Neurocan (R&D Systems), CS-56 (Sigma), 2B6 (Seikagaku) or anti-HS (HepSS-1), or biotinylated Wisteria Floribunda Agglutinin (WFA, Sigma) in the same solution. Following washes, sections were incubated with appropriate secondary antibodies: Alexa Fluor 488 goat anti-rabbit IgG or Alexa Fluor 568 anti-rabbit IgG (Life Technologies); FITC-conjugated goat F(ab’)2 anti-mouse IgM chain (Abcam); Alexa Fluor 488-streptavidin (Jackson Labs); Alexa Fluor 568-streptavidin (Life Technologies). Sections were mounted on silanated slides (KD Medical) and coverslipped using Fluoromount™ (Sigma) with or without DAPI. Confocal images were taken using a Zeiss LSM 510 UV microscope. 3-D co-localization analysis of confocal z-stacks was performed using routines incorporated in Imaris (Bitplane).
Yi J.H., Katagiri Y., Yu P., Lourie J., Bangayan N.J., Symes A.J, & Geller H.M. (2014). Receptor protein tyrosine phosphatase σ binds to neurons in the adult mouse brain. Experimental neurology, 255, 12-18.
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8

Visualizing Inflammasomes in F. novicida-Infected BMDMs

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F. novicida strain U112 or a strain expressing GFP was used for infection. For visualization of inflammasomes, BMDMs were infected for 20 h and further incubated in fresh media containing 1× FAM FLICA active caspase-1 for 1 h at 37°C (ImmunoChemistry Technologies). Cells were washed 3 times with PBS and fixed in 4% paraformaldehyde for 15 min at room temperature, followed by blocking in 10% normal goat serum (Dako) supplemented with 0.1% saponin (Sigma) for 1 h. Cells were incubated with a rabbit anti-ASC antibody (clone AL177, 1:500 dilution, AG-25B-0006-C100, AdipoGen) for 50 min at 37°C. For GBP5 staining, BMDMs were infected for the indicated times and washed 3 times with PBS. Cells were fixed and blocked as described above and stained using a rabbit anti-GBP5 antibody (1:500 dilution, 13220-1-AP, Proteintech) overnight at 4°C. The secondary antibody used was an Alexa Fluor 568 anti-rabbit IgG (Life technologies). Cells were counterstained in DAPI mounting medium (Vecta Labs). Bacteria, inflammasomes and BMDMs were visualized, counted, and imaged using a Nikon C2 confocal microscope.
Man S.M., Karki R., Malireddi R.K., Neale G., Vogel P., Yamamoto M., Lamkanfi M, & Kanneganti T.D. (2015). The transcription factor IRF1 and guanylate-binding proteins target AIM2 inflammasome activation by Francisella infection. Nature immunology, 16(5), 467-475.
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9

Visualizing Inflammasomes in F. novicida-Infected BMDMs

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F. novicida strain U112 or a strain expressing GFP was used for infection. For visualization of inflammasomes, BMDMs were infected for 20 h and further incubated in fresh media containing 1× FAM FLICA active caspase-1 for 1 h at 37°C (ImmunoChemistry Technologies). Cells were washed 3 times with PBS and fixed in 4% paraformaldehyde for 15 min at room temperature, followed by blocking in 10% normal goat serum (Dako) supplemented with 0.1% saponin (Sigma) for 1 h. Cells were incubated with a rabbit anti-ASC antibody (clone AL177, 1:500 dilution, AG-25B-0006-C100, AdipoGen) for 50 min at 37°C. For GBP5 staining, BMDMs were infected for the indicated times and washed 3 times with PBS. Cells were fixed and blocked as described above and stained using a rabbit anti-GBP5 antibody (1:500 dilution, 13220-1-AP, Proteintech) overnight at 4°C. The secondary antibody used was an Alexa Fluor 568 anti-rabbit IgG (Life technologies). Cells were counterstained in DAPI mounting medium (Vecta Labs). Bacteria, inflammasomes and BMDMs were visualized, counted, and imaged using a Nikon C2 confocal microscope.
Man S.M., Karki R., Malireddi R.K., Neale G., Vogel P., Yamamoto M., Lamkanfi M, & Kanneganti T.D. (2015). The transcription factor IRF1 and guanylate-binding proteins target AIM2 inflammasome activation by Francisella infection. Nature immunology, 16(5), 467-475.
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10

Spinal Cord Cell Isolation and Analysis

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Dissected and unfixed spinal cords were homogenized and submitted to consecutive washes with 0.1 M PBS pH 7.8 (n = 7 embryos per group of three independent experiments). Then, the cells were dissociated using 0.25% trypsin for 15 min at 37°C and added to 5% FBS under agitation. Subsequently, the samples were centrifuged at 640 ×g for 10 min and the supernatant was collected [30 (link)], incubated with primary antibodies rabbit anti-cyclin E IgG (1 : 1000, Santa Cruz Biotechnology, USA), mouse anti-p21 IgG (1 : 1000, Santa Cruz Biotechnology, USA), and mouse anti-βIII tubulin IgG (1 : 1000, Promega, USA) for 1 h, and then incubated for 45 min with the secondary antibodies Alexa Fluor 568 anti-rabbit IgG and Alexa Fluor 488 anti-mouse IgG (1 : 1000, Life Technologies, USA). Analyses were separately conducted for each antibody in each treatment. Previously, a run with unstained cells was performed to determine the gates of cells of interest. Additionally, propidium iodide was used to refine the gates of interest. Thus, from the dot plot with 20,000 events and considering the parameters side scatter (SSC-A) and forward scatter (FSC-A), the gates with 1,800 events were determined. FACSCanto II flow cytometer (BD Biosciences, Canada) was used for the analysis. The values are presented in absolute count.
Ferreira F.F., Ammar D., Bourckhardt G.F., Kobus-Bianchini K., Müller Y.M, & Nazari E.M. (2015). MeHg Developing Exposure Causes DNA Double-Strand Breaks and Elicits Cell Cycle Arrest in Spinal Cord Cells. Journal of Toxicology, 2015, 532691.
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