Prior to stimulation, neuronal cells were serum starved overnight in supplemented DMEM with 1% (v/v) N1 (30 (link)) at 37°C. Neurons were then stimulated for 15 min with PBS, G-CSF (200 ng/ml), or PMA (2 μM; Sigma-Aldrich). Cells were washed with ice-cold PBS, fixed (4% paraformaldehyde) for 20 min, incubated in methanol (100%, −20°C, 20 min), blocked, and permeabilized (0.01% Triton X-100, 5% FBS, and 5% goat serum in PBS, 60 min). Neurons were then stained with mouse anti-mouse NeuN (clone A60; Millipore) and rabbit anti-mouse phospho-p44/42 MAPK (Erk1/2, clone 197G2; Cell Signaling Technology) mAbs followed by Alexa Fluor 568–conjugated anti-mouse IgG (Thermo Fisher Scientific) and Alexa Fluor 488–conjugated anti-rabbit IgG, F(ab′)2 fragment (Thermo Fisher Scientific). Cells were stained with DAPI (1 μg/ml, 5 min; EMD Millipore) and coverslipped. Slides were examined using a Zeiss Axioskop 2 at ×10 magnification, and images were captured by a Zeiss AxioCam MRm. The percentage of neurons positive for phospho-ERK1/2 was calculated relative to the total NeuN-positive neuronal population.
Alexa fluor 568 conjugated anti mouse igg
Manufactured by Thermo Fisher Scientific
Sourced in United States
Alexa Fluor 568-conjugated anti-mouse IgG is a secondary antibody labeled with the Alexa Fluor 568 fluorescent dye. It is designed to detect and visualize mouse immunoglobulin G (IgG) in various immunoassays and imaging applications.
Automatically generated - may contain errors
Lab products found in correlation
Alexa fluor 488 conjugated anti rabbit igg, by Thermo Fisher Scientific (10 mentions)
Alexa fluor 488 conjugated anti mouse igg, by Thermo Fisher Scientific (6 mentions)
Hoechst 33342, by Thermo Fisher Scientific (5 mentions)
Alexa fluor 568 conjugated anti rabbit igg, by Thermo Fisher Scientific (3 mentions)
Ripa buffer solution, by Thermo Fisher Scientific (2 mentions)
Tcs sp8, by Leica (2 mentions)
Volocity software, by PerkinElmer (2 mentions)
Lsm 510 meta, by Zeiss (2 mentions)
Lsm 710, by Zeiss (2 mentions)
Axioskop 2, by Zeiss (1 mentions)
Axiocam mrm, by Zeiss (1 mentions)
G csf, by Merck Group (1 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Anti hmgb1, by Abcam (1 mentions)
Anti kshv orf 50, by Bioss Antibodies (1 mentions)
Eclipse e400 microscope, by Nikon (1 mentions)
Hep 2 human epithelial cell substrate slides, by Inova Diagnostics (1 mentions)
Inverted wide field fluorescence microscope, by Zeiss (1 mentions)
Formaldehyde, by Nacalai Tesque (1 mentions)
33 protocols using alexa fluor 568 conjugated anti mouse igg
1
Quantifying Neuronal Activation via Phospho-ERK1/2
2
Immunofluorescence Assay for HMGB1, KSHV Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunofluorescence assay (IFA) was performed as previously described (Lee M. S. et al., 2014 (link); Kang et al., 2021a (link)). The primary antibodies used were anti-HMGB-1 (Abcam), anti-KSHV ORF50 (Bioss), anti-KSHV K8.1 (Santa Cruz), and anti-LNA (Abcam) antibodies. The secondary antibodies used were Alexa Fluor 568-conjugated anti-mouse IgG (Thermo Fisher Scientific) and Alexa Fluor 568-conjugated anti-rabbit IgG (Thermo Fisher Scientific) antibodies. A concentration of 500 ng/mL of 49,6-diamidino-2-phenylindole (DAPI) was used to stain nucleic acids. The stained samples were observed under a Nikon Eclipse E400 microscope (Nikon, Tokyo, Japan) under the same conditions.
3
Quantification of Mouse ANA by IFA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
ANA in mouse sera were detected using Alexa Fluor568-conjugated anti-mouse IgG (ThermoFisher Scientific) and HEp-2 human epithelial cell substrate slides (INOVA Diagnostics) following the manufacturer’s instructions. The stained samples were examined with inverted wide-field fluorescence microscope (Zeiss) at a magnification of 400X. Images presented were processed using Zeiss software (ZEN blue edition). Quantification of the fluorescence intensity was performed using ImageJ 1.50i software as described. Integrated density measurement was performed to determine the fluorescence intensity of the staining.
4
Immunocytochemical Analysis of D2DR in Human Sperm
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Immunocytochemistry was performed to investigate D2DR localization in human sperm. The sperm samples from three patients were added to 2 ml phosphate‐buffered saline (PBS, pH 7.4), and the suspension was collected after centrifugation (1500× g for 5 min at RT). The sperm were fixed with 10% formaldehyde (Nacalai Tesque) for 15 min at 4°C, washed with PBS, and stored at 4°C. The sperm samples were collected by centrifugation (3890× g for 5 min at 4°C) and permeabilized with 1% Triton X‐100 in PBS for 15 min at 4°C. They were washed twice with PBS and blocked with 1% BSA in PBS for 60 min at 4°C. The sperm were incubated with mouse monoclonal anti‐D2DR antibody (1:50) overnight at 4°C. After three washes with PBS, the sperm were incubated (1 h at RT) with Alexa Fluor 568‐conjugated anti‐mouse IgG (1:200; Thermo Fisher Scientific), MitoTracker® Red CMXRos (250 nM, Thermo Fisher Scientific), and Hoechst 33342 (1:2000; Thermo Fisher Scientific). Treated samples were then washed with PBS, mounted on glass slides, and covered with coverslips. Images were obtained using an FV3000 confocal laser scanning microscope (Olympus).
5
Immunofluorescence Analysis of Myogenic Cells
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
C2C12 myotubes and hiPSC-myocytes (both differentiated in a 35-mm film bottom dish) were fixed and permeabilized as above. After blocking with 5% FBS in PBS for 1 h, specimens were incubated with anti-MHC antibody (1:100 dilution, R&D Systems) for 1 h followed by incaubation with Alexa Fluor 568-conjugated anti-mouse IgG (1:800 dilution, Thermo Fisher) for 45 min. Nuclei were stained with DAPI, and specimens were mounted as above. Cell images were obtained by an LSM800 confocal laser microscope (Carl Zeiss).
6
Reagents for Neuroscience Research
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Rabbit polyclonal anti-SESTD1 antibody was purchased from ProSci (Flint Place Poway, CA). Monoclonal anti-PSD-95, anti-β-actin and anti-Rac1 antibodies, and Rac1 activation assay kit were purchased from Millipore (Temecula, CA). Rabbit polyclonal anti-GFP and anti-synaptophysin antibodies were purchased from Abcam (Cambridge, MA). Monoclonal anti-bassoon antibody was purchased from Stressgen Bioreagents (Victoria, BC, Canada). Alexa-Fluor-488-conjugated anti-rabbit IgG and Alexa-Fluor-568-conjugated anti-mouse IgG were purchased from Molecular Probes (Eugene, OR). Rabbit polyclonal anti-Trio8 antibody was raised against recombinant Trio8 protein and was generated by LTK Biolaboratories (Taipei, Taiwan). Poly-L-lysine and bovine trypsin were purchased from Sigma-Aldrich (St. Louis, MO). Neurobasal-A, B-27 supplement, penicillin-streptomycin, L-glutamine and Lipofectamine 2000 were purchased from Invitrogen (Carlsbad, CA). T4 DNA ligation kit and restriction enzymes were purchased from New England Biolabs (UK). RIPA buffer solution was purchased from Thermo scientific (Rockford, IL).
7
Immunofluorescence Localization of Cellular Proteins
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
MA-10 cells grown on coverslips were rinsed in ice-cold phosphate buffered saline (PBS) and fixed with 4% paraformaldehyde at room temperature for 20 min, or with cold methanol at –20 °C for 10 min. After being washed with cold PBS, fixed cells were permeabilized and blocked with a blocking buffer (5% normal goat serum in 20 mM phosphate buffer, pH 7.4, 0.1% (v/v) Triton X-100, and 0.45 M NaCl) for 60 min at 4 °C. Cells were then immunolabeled overnight at 4 °C with 1:200 dilution (in the blocking buffer) of rabbit anti-Aha1, mouse anti-ClC-2 (Santa Cruz, Dallas, TX, USA), rabbit anti-FKBP8, rabbit anti-HOP, rabbit anti-Hsc70, or rabbit anti-Hsp90β antibodies. Alexa Fluor 488-conjugated anti-rabbit IgG and Alexa Fluor 568-conjugated anti-mouse IgG (1:200; Molecular Probes, Eugene, OR, USA) were used as secondary antibodies. Nuclei were stained with DAPI. After final wash, coverslips were mounted in a mounting medium (4% n-propylgallate, 90% glycerol, 0.1 M carbonate buffer, pH 9.2). Fluorescence images were viewed and acquired with a confocal microscope (TCS SP8, Leica, Wetzlar, Germany).
8
Antibody Characterization for Viral Protein Detection
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The rabbit polyclonal antibodies recognizing the C terminus of IBV E and the N-terminal head of golgin-160 have been previously described (42 (link), 43 (link)). The antibody generated against the C terminus of IBV S (anti-IBV SCT) has also been previously described (26 (link)). The mouse monoclonal antibody recognizing IBV S1 was a gift from Ellen Collisson (25 (link)). The mouse monoclonal antibody recognizing the N terminus of influenza A virus M2 has been previously described and was a generous gift from Andrew Pekosz (44 (link)). The mouse anti-GFP antibody was obtained from Roche (Mannheim, Germany). The rabbit anti-GFP antibody was obtained from Thermo Fisher Scientific (Rockford, IL). Alexa Fluor 488-conjugated anti-rabbit IgG, Alexa Fluor 488-conjugated anti-mouse IgG, Alexa Fluor 568-conjugated anti-rabbit IgG, and Alexa Fluor 568-conjugated anti-mouse IgG were obtained from Invitrogen/Molecular Probes (Eugene, OR).
9
Staining Malaria Parasite Flagella
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Before setting up ookinete cultures or mosquito infections, exflagellation was assessed as previously described (Ramakrishnan et al., 2013 (link)). To stain flagella with anti-α-tubulin, samples were fixed 15 min after inducing exflagellation in fresh 4% paraformaldehyde (PFA). The suspension was allowed to adhere onto poly-L-lysine-coated slides overnight at 4°C. The slides were washed once with PBS, and parasites were stained using the mouse monoclonal anti-α-tubulin II antibody (Sigma-Aldrich, T9026, 1:250). Secondary antibody Alexa Fluor 568-conjugated anti-mouse IgG for fluorescence detection was used at 1:1000 (Molecular Probes, Thermo Fisher Scientific, A-11008). The slides were mounted in Vectashield with DAPI (Vector Laboratories). Parasites were visualised on a Leica SP5 confocal microscope and acquired and analysed with the LAS AF Lite software (Leica).
10
Immunofluorescence Imaging of LRP-1 in Osteoarthritis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
OA and normal cartilage samples (n = 3 each) were rested in culture with DMEM for 2 days. Explants were then snap‐frozen and sectioned (5‐µm sections) using a CM1900 cryostat (Leica Microsystems). Each sample was fixed with methanol and incubated with anti–LRP‐1 β‐chain antibody for 3 hours at room temperature. Incubation with Alexa Fluor 568–conjugated anti‐mouse IgG (Molecular Probes) for 1 hour at room temperature was used to visualize the antigen signals. Nuclei were stained with DAPI. Samples were viewed using an Eclipse TE2000‐U confocal laser scanning microscope (Nikon). Data were collated using Volocity software (Improvision).
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!