Alexa fluor 488 goat anti chicken

Manufactured by Thermo Fisher Scientific

Sourced in United States

Alexa Fluor 488 goat anti-chicken is a fluorescent secondary antibody used to detect and visualize chicken-derived antigens in various immunoassay techniques. It is conjugated with the Alexa Fluor 488 dye, which emits green fluorescence upon excitation.

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Alexa Fluor 488 (6806 citations) Alexa 488 (1863 citations) Goat anti-chicken Alexa 488 (45 citations) Alexa Fluor 488 goat anti-chicken IgG (41 citations) Goat anti-chicken 488 (30 citations) Alexa Fluors (21 citations) Goat anti-Chicken IgY-Alexa Fluor 488 (16 citations) Alexa Fluor 488-conjugated goat anti-chicken (15 citations) Alexa Fluor 488 anti-chicken (10 citations) Alexa 488 anti-chicken (10 citations)

101 protocols using alexa fluor 488 goat anti chicken

Lung Tissue Immunofluorescence Staining

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Mouse lungs were perfused and inflated with 20 ml/kg aqueous buffered zinc formalin (Z-FIX; Anatech, Battle Creek, MI) immediately following euthanasia and used for paraffin-embedding. Immunofluorescence staining was performed as described previously (Lee et al., 2017 (link)). Paraffin-embedded tissue sections (5 μm) were rehydrated and subjected to antigen retrieval in Tris-HCl buffer (100 mM, pH 9.5). Blocking was performed with 15% BSA in 0.2% Triton-X/PBS at room temperature for 60 min. Sections were incubated with chicken anti-GFP (1:500, Abcam, ab13970) and goat anti-SP-C (1:50, Santa Cruz Biotechnology, sc-7706) overnight at 4°C. The immune complexes were detected using Alexa Fluor 568 donkey anti-goat or Alexa Fluor 488 goat anti-chicken (1:400, Invitrogen) secondary antibodies before sections were counterstained with DAPI. Sections were mounted in Prolong Gold antifade reagent (Invitrogen). Fluorescence images were acquired using a Zeiss microscope.

Zhu B., Wu Y., Huang S., Zhang R., Son Y.M., Li C., Cheon I.S., Gao X., Wang M., Chen Y., Zhou X., Nguyen Q., Phan A.T., Behl S., Taketo M.M., Mack M., Shapiro V.S., Zeng H., Ebihara H., Mullon J.J., Edell E.S., Reisenauer J.S., Demirel N., Kern R.M., Chakraborty R., Cui W., Kaplan M.H., Zhou X., Goldrath A.W, & Sun J. (2021). Uncoupling of macrophage inflammation from self-renewal modulates host recovery from respiratory viral infection. Immunity, 54(6), 1200-1218.e9.

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Visualizing rAAV-Mediated Gene Expression

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Immunohistochemistry (IHC) was performed on free-floating sections through injected brain regions in order to visualize the location and spread of rAAV-mediated expression, in addition to providing an estimation of transduction efficiency. Sections were washed in 0.05M phosphate buffer containing 0.1% Triton x-100 (PB-TX) and incubated overnight in either 1:2000 rabbit antiTH (Millipore AB152), 1:1000 chicken anti-GFP (Abcam 13970) or 1:2000 goat anti-FLAG (Abcam 1257). Following overnight incubation in primary antibody, sections were washed in PB-TX and incubated in either 1:500 Cy-3 conjugated goat anti-rabbit (Jackson immune research 111-165-003), 1:1000 Alexafluor 488 donkey anti-goat (Invitrogen A11055) or 1:1000 Alexafluor 488 goat anti-chicken (Invitrogen A11039) secondary antibodies for 1h at room temperature and protected from light. Sections were then washed and covered slipped using ProLong Gold antifade reagent (Molecular Probes, Eugene, OR). Sections were viewed on a Nikon TE-2000-U-Fluorescence Microscope (Melville, NY)

Benskey M.J., Manfredsson F.P., Lookingland K.J, & Goudreau J.L. (2014). The Role of Parkin in the Differential Susceptibility of Tuberoinfundibular and Nigrostriatal Dopamine Neurons to Acute Toxicant Exposure. Neurotoxicology, 46, 1-11.

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Immunostaining of Zebrafish Embryos

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Embryos for most antibodies were fixed at the indicated stage in 4% paraformaldehyde, permeabilized in PBST (PBS+0.5% Triton X-100), and blocked in PBST+2% bovine serum albumin. Antibodies and concentrations were as follows: anti-Pax2a (GTX128127, Genetex; 1:200); anti-pSmad3 (ab52903, Abcam; 1:200); anti-Laminin 1 (L9393, Sigma-Aldrich; 1:100); anti-fibronectin (F3648, Sigma-Aldrich; 1:100); anti-GFP (A10262, Invitrogen; 1:200).
In accordance with methods described by Carrara et al. (2019) (link), anti-Nidogen 1/Entactin (ab14511, Abcam) was used at 1:100 and required a modified embryo preparation after 4% paraformaldehyde fixation: embryos were permeabilized in 30 μg/ml Proteinase K for 15 min, and blocked in 0.8% PBST+10% sheep serum+1% bovine serum albumin. The antibody was applied in 0.8% PBST+1% sheep serum+1% bovine serum albumin.
Secondary antibodies used were: Alexa Fluor 488 goat anti-mouse (A-11001, Invitrogen), Alexa Fluor 488 goat anti-rabbit (A-11008, Invitrogen), Alexa Fluor 488 goat anti-chicken (A-11039, Invitrogen), all used at 1:200. Nuclei were detected by incubation with 1 µM TO-PRO-3 iodide (T3605, Invitrogen). Embryos were cleared through a series of 30%/50%/70% glycerol (in PBS) prior to imaging.

Bryan C.D., Casey M.A., Pfeiffer R.L., Jones B.W, & Kwan K.M. (2020). Optic cup morphogenesis requires neural crest-mediated basement membrane assembly. Development (Cambridge, England), 147(4), dev181420.

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Immunostaining Protocols for Drosophila Eye Imaginal Discs

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Wandering 3rd instar larvae were collected from respective genetic crosses and eye imaginal discs were processed for immunostaining as previously described (Sudhakaran et al. 2014 (link)). Each immunostaining experiment was reproduced in at least three independent biological replicates: for each replicate, discs dissected from multiple larvae were individually examined.
Primary antibodies used were: rabbit anti-RFP (Invitrogen R103367,) at 1:1000, rabbit-anti-Rasputin (Aguilera-Gomez et al. 2017 (link)) 1:1000, rabbit-anti-Caprin (Papoulas et al. 2010 (link)) at 1:1000, rat-anti-Elav-7E8A10 (Developmental Studies Hybridoma Bank) 1:300 and chicken-anti-Ataxin-2 (Bakthavachalu et al. 2018 (link)) at 1:500. Secondary antibodies were used at 1:1000 dilution: Alexa Fluor^®488 goat anti-Chicken (Invitrogen A11039), Alexa Fluor^®488 goat anti-Rabbit (Invitrogen A11078), Alexa Fluor^®488 goat anti-Rat (Invitrogen A11006), and Fluor^®555 goat anti-Rabbit (Invitrogen A21428).
Prepared eye-imaginal discs were mounted in Vectashield Mounting Medium (Vector Labs, H-1000) on microscope slides and imaged on a Zeiss LSM880 confocal microscope.

Huelsmeier J., Walker E., Bakthavachalu B, & Ramaswami M. (2021). A C-terminal ataxin-2 disordered region promotes Huntingtin protein aggregation and neurodegeneration in Drosophila models of Huntington’s disease. G3: Genes|Genomes|Genetics, 11(12), jkab355.

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Immunofluorescence Staining of Mouse Lungs

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Mouse lungs were perfused and inflated with 20 ml/kg aqueous buffered zinc formalin (Z-FIX; Anatech, Battle Creek, MI) immediately following euthanasia and used for paraffin-embedding. Immunofluorescence staining was performed as described previously (Lee et al., 2017 (link)). Paraffin-embedded tissue sections (5 μm) were rehydrated and subjected to antigen retrieval in Tris-HCl buffer (100 mM, pH 9.5). Blocking was performed with 15% BSA in 0.2% Triton-X/PBS at room temperature for 60 min. Sections were incubated with chicken anti-GFP (1:500, Abcam, ab13970) and goat anti-SP-C (1:50, Santa Cruz Biotechnology, sc-7706) overnight at 4 °C. The immune complexes were detected using Alexa Fluor 568 donkey anti-goat or Alexa Fluor 488 goat anti-chicken (1:400, Invitrogen) secondary antibodies before sections were counterstained with DAPI. Sections were mounted in Prolong Gold antifade reagent (Invitrogen). Fluorescence images were acquired using a Zeiss microscope.

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Immunostaining of Hippocampal Sections

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Slide mounted brain sections (two per slide) from dorsal hippocampus were processed for histological studies. Sections were immunostained with the following antibodies: rabbit anti-pS6 (1:500, Cell Signaling), chicken anti-GFP (1:500, Abcam, Boston, MA), rabbit anti-zinc transporter 3 (ZnT3) (1:3000, Synaptic Systems, Gottingen, Germany), rabbit anti-Glutamate receptor 2/3 (1:100, Millipore, Temecula, CA), rabbit anti-prox1 (1:1000, Sigma-Aldrich, St. Louis, MO), rabbit anti-GFAP (1:500, Millipore) and rabbit anti-Iba1 (1:1000, Synaptic Systems). AlexaFluor488 goat anti-chicken, AlexaFluor488 goat anti-rabbit, AlexaFluor594 goat anti-rabbit and AlexaFluor647 goat anti-rabbit secondary antibodies were used (Invitrogen, Eugene, OR). Sections were dehydrated in alcohols, cleared in xylenes and coverslips were attached with Krystalon mounting-media (Harleco, Darmstadt, Germany).

Hester M.S., Hosford B.E., Santos V.R., Singh S.P., Rolle I., LaSarge C.L., Liska J.P., Garcia-Cairasco N, & Danzer S.C. (2016). Impact of rapamycin on status epilepticus induced hippocampal pathology and weight gain. Experimental neurology, 280, 1-12.

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Brain Immunostaining and Imaging

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Adult heads were fixed with 4% formaldehyde (Thermo Scientific, #28906) in 1x PBS and 0.1% Triton X-100 (PTX) for 30 min and washed 5 times, 2 minutes each with PTX before dissection. Brains were dissected in PTX. Dissected brains were subsequently fixed in 4% formaldehyde in PTX for 10 min and washed 5 times, 2 minutes each with PTX. Primary antibody anti-chicken GFP (Aves Labs, #GFP-1020) was used at 1:500 in PTX and rocked overnight at 4°C. Brains were washed 3 times, 10 minutes each with PTX. Alexa Fluor 488 goat anti-chicken (Invitrogen, #A11039) was used at 1:100 in PTX and rocked overnight at 4°C. Following antibody incubation brains were washed 3 times, 10 minutes each with PTX. Brains were mounted in ProLong Gold with DAPI (Invitrogen, #P36935).

Herrmann K.A., Liu Y., Llobet-Rosell A., McLaughlin C.N., Neukomm L.J., Coutinho-Budd J.C, & Broihier H.T. (2022). Divergent signaling requirements of dSARM in injury-induced degeneration and developmental glial phagocytosis. PLoS Genetics, 18(6), e1010257.

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Immunofluorescence Analysis of Nephrocytes and Fat Bodies

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Immunofluorescence analyses of nephrocytes and fat bodies were performed as described (Lőrincz et al., 2016a (link); Takáts et al., 2013 (link)). The following antibodies were used: mouse anti-Rab7 (1:10, DSHB) (Riedel et al., 2016 (link)), rabbit anti-CathL (1:100, ab58991; Abcam), rat anti-Atg8a (1:300) (Takáts et al., 2013 (link)), rabbit anti p62/Ref(2)p 1:1000 (Pircs et al., 2012 (link)), rabbit anti-Rab5 (1:100, Abcam, ab31261), chicken anti-GFP (1:1500, Invitrogen: A10262), rabbit anti-HA 1:100 (Sigma-Aldrich: H6908), monoclonal rat anti-HA 1:80 (Roche, 3F10), rat anti-Rbsn-5 (1:1,000) (Tanaka and Nakamura, 2008 (link)). Alexa Fluor 568 goat anti-mouse, Alexa Fluor 568 goat anti-rat, Alexa Fluor 488 goat anti-rabbit, Alexa Fluor 488 goat anti-rat, Alexa Fluor 488 goat anti-chicken (all 1:1,000, Invitrogen).

Lőrincz P., Kenéz L.A., Tóth S., Kiss V., Varga Á., Csizmadia T., Simon-Vecsei Z, & Juhász G. (2019). Vps8 overexpression inhibits HOPS-dependent trafficking routes by outcompeting Vps41/Lt. eLife, 8, e45631.

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Fluorescent Protein Staining Protocol

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The primary antibody solution contained chicken anti-GFP (1:1000, Abcam) and mouse anti-Bruchpilot (1:30, Developmental Studies Hybridoma Bank, nc82). The secondary antibody solution contained Alexa Fluor 488 goat anti-chicken (1:250, Invitrogen) and Alexa Fluor 633 goat anti-mouse (1:250, Invitrogen).

Joubès J., Phan T.H., Just D., Rothan C., Bergounioux C., Raymond P, & Chevalier C. (1999). Molecular and Biochemical Characterization of the Involvement of Cyclin-Dependent Kinase A during the Early Development of Tomato Fruit. Plant Physiology, 121(3), 857-869.

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Immunostaining for GFP in Brain Slices

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Sections were rinsed by PBS and blocked by 2% Albumin Bovine – 0.3% Triton – PBS for 1 h. Then, slices were incubated with 1:500 chicken anti-GFP antibody (Invitrogen, A10262) overnight and followed by 1-h incubation with 1:500 Alexa Fluor 488 goat anti-chicken (Invitrogen, A11039) secondary antibody and DAPI. Slices were washed by PBS after incubation with antibodies. Finally, slices were mounted on slides and covered with mounting medium.

Zhang X., Cao Q., Gao K., Chen C., Cheng S., Li A., Zhou Y., Liu R., Hao J., Kropff E, & Miao C. (2024). Multiplexed representation of others in the hippocampal CA1 subfield of female mice. Nature Communications, 15, 3702.

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