Alexa fluor 488 donkey anti rabbit igg

Manufactured by Thermo Fisher Scientific

Sourced in United States, Germany, United Kingdom, France

Alexa Fluor 488 donkey anti-rabbit IgG is a secondary antibody conjugated with the Alexa Fluor 488 fluorescent dye. It is designed to detect and visualize rabbit immunoglobulin G (IgG) in various immunoassays and imaging applications.

Automatically generated - may contain errors

Lab products found in correlation

272 protocols using alexa fluor 488 donkey anti rabbit igg

Neuroglobin Expression in Ischemic Cortex

Check if the same lab product or an alternative is used in the 5 most similar protocols

The rats (N = 5 per group) were anesthetized with 10% chloral hydrate and were subsequently sacrificed by transcardiac perfusion with physiological saline followed by 4% paraformaldehyde dissolved in PBS (pH 7.4). The frozen sections (15μm thick) were incubated with rabbit polyclonal anti-NGB antibody (1:200 dilution) in PBS containing 0.03% Triton X-100 at 4°C overnight (Sun, et al., 2001 (link)). After washing, the sections were subsequently incubated with Alexa Fluor 488 donkey anti-rabbit IgG for 1 h at room temperature (Molecular Probes, 1:500). Slides were mounted using ProLong Gold anti-fade reagent with DAPI (Molecular Probes). The quantitative analysis was expressed using the average of four brain slides. Ngb-positive signal within the ipsilateral cortex peri-infarct region were examined and quantified using Image-Pro Plus software 5.0 in a blinded manner at 1, 7, and 14 days after reperfusion.

Ren C., Wang P., Wang B., Li N., Li W., Zhang C., Jin K, & Ji X. (2015). Limb remote ischemic per-conditioning in combination with post-conditioning reduces brain damage and promotes neuroglobin expression in the rat brain after ischemic stroke. Restorative Neurology and Neuroscience, 33(3), 369-379.

+ Open protocol

+ Expand

Multicolor Immunofluorescence Labeling Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

The selected slides were placed in an oven at 60°C to remove excess paraffin, dehydrated in a series of graded alcohols, and placed in a Coplin jar with 10 mM citrate buffer at pH 6, inside a pressure cooker for 20 min to perform antigen recovery. Then, the tissues were permeabilized with PBS-T (0.01 M PBS and 0.1% Triton) for 30 min. Non-specific sites were blocked with 5% horse serum in PBS-T in a humid chamber for 30 min, and the tissues were incubated with goat anti-GFAP (1:500, Abcam), rabbit anti-cleave caspase-3 (1:300, Cell Signaling), and mouse anti–βIII-tubulin (1:500, Abcam) primary antibodies in a humid chamber at 4°C overnight. Afterward, the tissues were washed with PBS-T and incubated with Alexa Fluor^® 488 donkey anti-rabbit IgG (1:500, Molecular Probes), Alexa Fluor^® 594 donkey anti-goat IgG (1:500, Molecular Probes), and Alexa Fluor^® 405 donkey anti-mouse IgG (1:500, Molecular Probes) secondary antibodies for 2 h at room temperature in the dark. Then slides were placed in a solution of 0.1% Black Sudan B (Sigma-Aldrich) in 70% ethanol for 15 min, the excess Black Sudan B was removed using distilled water and PBS-T. The tissues were covered with VECTASHIELD^® and a coverslip and were observed under a confocal Nikon TI eclipse microscope.

Coyoy-Salgado A., Orozco-Barrios C., Sánchez-Torres S., Olayo M.G., Cruz G.J., Morales-Corona J., Olayo R., Diaz-Ruiz A., Ríos C., Alvarez-Mejia L., Mondragón-Lozano R., Morales-Guadarrama A., Alonso-García A.L., Fabela-Sánchez O, & Salgado-Ceballos H. (2023). Gene expression and locomotor recovery in adult rats with spinal cord injury and plasma-synthesized polypyrrole/iodine application combined with a mixed rehabilitation scheme. Frontiers in Neurology, 14, 1124245.

+ Open protocol

+ Expand

Immunocytochemical Analysis of Autophagy Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were fixed in 4% paraformaldehyde for 20 min at room temperature and permeabilized with 0.2% Triton X-100 containing 1% bovine serum albumin (BSA) in phosphate-buffered saline (PBS) for 15 minutes. After blocking with 1% BSA, fixed cells were incubated overnight at 4°C with primary antibodies against LC3 (1:100 dilution; Cell Signaling, Danvers, MA, http://www.cellsignal.com/) or lysosomal-associated membrane protein-1 (LAMP-1, 1:200 dilution; Hybridoma Bank, Iowa City, IA, http://dshb.biology.uiowa.edu/). Stained cells were washed and incubated for 2 hours at room temperature with AlexaFluor-conjugated secondary antibodies (AlexaFluor 555-donkey anti–mouse IgG 1:500 dilution or AlexaFluor 488-donkey anti–rabbit IgG 1:500 dilution; Molecular Probes, http://www.invitrogen.com/). After incubation with secondary antibodies, cells were stained with nuclear dye DAPI (Life technologies, Grand Island, NY, www.lifetechnologies.com/) in PBS for 5 minutes at room temperature. For negative controls, cultured cells were incubated with secondary antibody only.

Ou X., Lee M.R., Huang X., Messina-Graham S, & Broxmeyer H.E. (2014). SIRT1 positively regulates autophagy and mitochondria function in embryonic stem cells under oxidative stress. Stem cells (Dayton, Ohio), 32(5), 1183-1194.

+ Open protocol

+ Expand

Immunohistochemical Analysis of c-Fos and OXT

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Serial 40-μm-thick sections were rinsed twice with 0.1 M phosphate-buffered saline (PBS) and washed in 0.1 M Tris buffer (pH, 7.6) containing 0.3% Triton X-100. Sections were incubated for 72 h at 4 °C in primary antibody solution (goat anti-c-Fos, Santa Cruz Biotechnology, TX, USA; 1:500 or rabbit anti-OXT, Sigma-Aldrich, MO, USA; 1:5000)^{65 (link)}. After washing twice in 0.3% Triton X-100 in PBS, floating sections were incubated for 24 h at 4 °C with a secondary antibody (Alexa Fluor 546 donkey anti-goat IgG or Alexa Fluor 488 donkey anti-rabbit IgG; Molecular Probes, OR, USA; 1:2,000 in PBS containing 0.3% Triton X-100)^{65 (link)}. Sections were washed twice in PBS and then mounted on the slides and coverslipped using vectashield (Vector Laboratories Co. Ltd., CA, USA)^{66 (link)}. Images of Fos⁺, OXT⁺, and Fos⁺/OXT⁺ double-labelled cells were counted manually by two researchers who were blinded to avoid bias. The number and percentage of Fos⁺, OXT⁺, and Fos⁺/OXT⁺ cells in the SON and PVN were estimated.

Nishimura K., Yoshino K., Ikeda N., Baba K., Sanada K., Akiyama Y., Nishimura H., Tanaka K., Sonoda S., Ueno H., Yoshimura M., Maruyama T., Hachisuga T, & Ueta Y. (2022). Oestrogen-dependent hypothalamic oxytocin expression with changes in feeding and body weight in female rats. Communications Biology, 5, 912.

+ Open protocol

+ Expand

Niclosamide Sensitizes TNBC Cells to IR

Check if the same lab product or an alternative is used in the 5 most similar protocols

TNBC cells growing on coverslips were treated with niclosamide (1.5 μM) for 18 h. The cells were then exposed to 6 Gy γ-ray IR. After 6 h incubation, the cells were fixed in 4% paraformaldehyde, permeabilized with 0.5% Triton X-100, blocked with 10% (v:v) FBS in phosphate-buffered saline (PBS), labeled with rabbit monoclonal β-catenin (6B3) antibody (1:200, Cell Signaling Technology), and detected with Alexa Fluor 488 donkey anti-rabbit IgG (1:500, Molecular Probes/ThermoFisher Scientific, Grand Island, NY). The nuclei were counter stained with DAPI (Santa Cruz Biotechnology Inc., Santa Cruz, CA), and the images were examined under an Olympus BX51 fluorescent microscope. The mean fluorescence intensity of nuclear β-catenin staining was calculated out of a total number of 200 cells per sample, and was normalized to that of a control sample for the calculation of the relative fluorescence intensity.

Yin L., Gao Y., Zhang X., Wang J., Ding D., Zhang Y., Zhang J, & Chen H. (2016). Niclosamide sensitizes triple-negative breast cancer cells to ionizing radiation in association with the inhibition of Wnt/β-catenin signaling. Oncotarget, 7(27), 42126-42138.

+ Open protocol

+ Expand

NF-κB p65 Translocation Assay

Check if the same lab product or an alternative is used in the 5 most similar protocols

FLS cells were cultivated in 96-well plates (at 1 × 10⁴/ml) at 37°C for 12 h. Ebosin (80 μg/ml) was added in each well followed by cultivation at 37°C for 12 h and then treated cell with TNF-α (10 ng/ml) for an additional 3 h. The cells were fixed in 5% paraformaldehyde (PFA)/PBS for 10 min at room temperature, permeabilized with PBS/0.1% Triton-X100 for 15 min, and blocked in PBS with 5% bovine serum albumin for 1 h. The cells were incubated with rabbit anti-NF-κB p65 (Cell Signaling Technology) and Alexa Fluor 488 donkey anti-rabbit IgG (Molecular Probes). The fluorescent signals were detected by fluorescence microscopy (Olympus IX71, Japan).

Zhang Y., Wang L., Bai L., Jiang R., Wu J, & Li Y. (2022). Ebosin Attenuates the Inflammatory Responses Induced by TNF-α through Inhibiting NF-κB and MAPK Pathways in Rat Fibroblast-Like Synoviocytes. Journal of Immunology Research, 2022, 9166370.

+ Open protocol

+ Expand

Imaging Bacterial Biofilms with CLSM

Check if the same lab product or an alternative is used in the 5 most similar protocols

CLSM was performed as previously described with some modifications (Armbruster, et al., 2010 ). S. pneumoniae EF3030 and/or M. catarrhalis O35E were seeded in 4 well Permanox chamber slides (Thermo Scientific) as previously stated and grown for 24 hours at 37°C in supplemented TSB. The biofilms were fixed (0.3% paraformaldehyde), frozen in embedding medium (Tissue-Tek), and cryosectioned laterally (~5 μm per section). Immunofluorescent staining was performed using rabbit polyclonal antiserum against pneumococcal surface protein A (PspA) and monoclonal antibody Mab 3F5-5E5 which recognizes a conserved M. catarrhalis surface epitope (Furano, et al., 2005 (link)) along with appropriate fluorescent secondary antibody conjugates (S. pneumoniae: Alexa Fluor 488 donkey anti-rabbit IgG, M. catarrhalis: Texas Red goat anti-mouse IgM) (Molecular Probes). Microscopy was performed using a Nikon Eclipse confocal laser scanning microscope. Images were analyzed using the COMSTAT program within MatLab 7.0.4 software.

Perez A.C., Pang B., King L.B., Tan L., Murrah K.A., Reimche J.L., Wren J.T., Richardson S.H., Ghandi U, & Swords W.E. (2014). Residence of Streptococcus pneumoniae and Moraxella catarrhalis within polymicrobial biofilm promotes antibiotic resistance and bacterial persistence in vivo. Pathogens and disease, 70(3), 280-288.

+ Open protocol

+ Expand

Antibody Panel for Protein Detection

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following antibodies were used in immunostaining or western blotting: rabbit anti-TTR (A0002, Agilent Dako, Santa Clara, CA), rabbit anti-beta Amyloid 1–42 antibody [mOC98] (ab201061, abcam, Cambridge, MA), rabbit anti-α-syn (ab131508, abcam, Cambridge, MA), SERPINA1 (ARP59239_P050, Aviva Systems Biology, San Diego, CA), mouse anti-β-actin (Cell Signaling Technology), goat anti-rabbit or mouse HRP-conjugated IgG (Cell Signaling Technology), and Alexa Fluor 488 donkey anti-rabbit IgG (Molecular Probes, A-21206).

Cheng S., Banerjee S., Daiello L.A., Nakashima A., Jash S., Huang Z., Drake J.D., Ernerudh J., Berg G., Padbury J., Saito S., Ott B.R, & Sharma S. (2021). Novel blood test for early biomarkers of preeclampsia and Alzheimer’s disease. Scientific Reports, 11, 15934.

+ Open protocol

+ Expand

Immunofluorescent Staining of Hair Cells

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Immunofluorescent staining procedures were performed as antecedent description [7 (link)]. The primary antibodies were rabbit anti-prestin, (sc-30163, Santa Cruz), goat anti-SPAG6 (sc-165529, Santa Cruz) and rabbit anti-myosinVIIa (PA1-936, Thermo Scientific Pierce Antibodies). The secondary antibodies were Alexa Fluor 488 donkey anti-rabbit IgG (A-21206, Molecular Probes) and Alexa Fluor 647 donkey anti-goat IgG (A-21447, Molecular Probes). The cell nucleus and the F-actins (cilia bundles) were stained by DAPI (D9542, SIGMA) and FITC-Phalloidin (P5282, SIGMA), respectively.
Specimens were observed under a laser scanning confocal microscope (TSC SPE, LEICA). The 488 nm laser was used for the visualization of Alexa Fluor 488 and FITC-Phalloidin staining. The 635 nm laser was used for the visualization of Alexa Fluor 647. DAPI staining was watched under UV light, the 405 nm laser.
For hair cells counting, we used the cell counter tool in Image J software to accumulate the myosinVIIa-positive hair cells within the 400 μm length in the middle turn of the cochlea [17 (link)]. As for the quantification of fluorescence intensity of prestin, we also performed as previous study [17 (link)]. The fluorescence intensity ratio of Spag6 −/− to Spag6 +/+ mice in different time points were calculated.

Wang J., Li X., Zhang Z., Wang H, & Li J. (2015). Expression of prestin in OHCs is reduced in Spag6 gene knockout mice. Neuroscience letters, 592, 42-47.

+ Open protocol

+ Expand

Immunofluorescence Staining of Mouse Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

Rat anti-mouse CD54 (intracellular adhesion molecule 1 (ICAM-1)) antibody (clone #: YN1/1.7.4, rat IgG_{2b, κ}) was purchased from BioLegend (San Diego, CA, U.S.A.). Rat anti-mouse CD45 (clone #: 30-F11) was obtained from BD Biosciences Pharmingen (San Diego, CA, U.S.A.). Rabbit anti-GFP (ab6556) was purchased from Abcam (Cambridge, MA, U.K.). Alexa Fluor® 594 Donkey anti-Rat IgG, Alexa Fluor®488 Donkey anti-Rabbit IgG and Alexa Fluor®488 phalloidin were sourced from Molecular Probes (Invitrogen, U.S.A.). Recombinant mouse IL-1β and MCP-1 were purchased from PROSPEC (East Brunswick, NJ, U.S.A.). Recombinant human fibroblast growth factor 2 (rhFGF-2) was obtained from Miltenyi Biotec (Bergisch-Gladbach, Germany).

Ibi M., Horie S., Kyakumoto S., Chosa N., Yoshida M., Kamo M., Ohtsuka M, & Ishisaki A. (2018). Cell–cell interactions between monocytes/macrophages and synoviocyte-like cells promote inflammatory cell infiltration mediated by augmentation of MCP-1 production in temporomandibular joint. Bioscience Reports, 38(2), BSR20171217.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!