Cells were grown on four chamber CultureSlides (FALCON) and treated with 10 nmol/L ispinesib for 96 hours. The cells were fixed in 4% formaldehyde in PBS for 15 minutes, blocked with blocking buffer (PBS with 5% BSA and 0.2% Triton X‐100) for 1 hour, and incubated overnight with the primary antibody at 4℃ (anti–α‐tubulin 1:500, anti–Pericentrin 1:1000, anti–Cleaved Caspase 3; 1:500). Alexa Fluor 594‐conjugated goat anti–rabbit IgG antibodies and Alexa Fluor 488‐conjugated goat anti–mouse IgG antibodies (#8889 and #4408, respectively; Cell Signaling Technology) were used as secondary antibodies (1:1000). The cells were viewed under a fluorescence microscope (BZ‐700; KEYENCE and Leica DMi8; Leica microsystems).
Alexa fluor 594 conjugated goat anti rabbit igg antibodies
Manufactured by Cell Signaling Technology
Sourced in United States
Alexa Fluor 594‐conjugated goat anti–rabbit IgG antibodies are a fluorescent-conjugated secondary antibody used for detection and visualization of rabbit primary antibodies. The Alexa Fluor 594 dye provides a bright, photostable fluorescent signal.
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Lab products found in correlation
Dexamethasone (dex), by Santa Cruz Biotechnology (1 mentions)
Dihydroethidium dhe, by Thermo Fisher Scientific (1 mentions)
Phospho p38, by Cell Signaling Technology (1 mentions)
Hrp conjugated anti rabbit igg secondary antibody, by Merck Group (1 mentions)
Glyceraldehydes 3 phosphate dehydrogenase gapdh, by Cell Signaling Technology (1 mentions)
5 lox, by Cell Signaling Technology (1 mentions)
β actin, by Cell Signaling Technology (1 mentions)
2 protocols using alexa fluor 594 conjugated goat anti rabbit igg antibodies
1
Immunofluorescence Assay for Cellular Changes
2
Antibody-based Signaling Pathway Analysis
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Antibodies against ERK1/2, phospho-ERK1/2, Akt, phospho-Akt, c-Jun N-terminal kinases (JNK), phospho-JNK, p38, phospho-p38, COX-2, 5-LOX, glyceraldehydes-3-phosphate dehydrogenase (GAPDH), β-actin, lamin b, and Alexa Fluor 594 conjugated goat anti-rabbit IgG antibodies were obtained from Cell Signaling Technology (Boston, MA, USA). Anti-NF-κB and anti-mouse horseradish peroxidase- (HRP-) conjugated IgG antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Dihydroethidium (DHE) was purchased from molecular probes (Eugene, OR, USA). Dexamethasone was obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-rabbit HRP-conjugated IgG secondary antibody and other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA), unless otherwise indicated. Gallic acid-l-leucine (GAL) conjugate was synthesized from gallic acid and methyl-l-leucine hydrochloride by following the previous method as described [15 (link)]. The structure of GAL conjugate was confirmed by NMR technique whereas the purity of the product was determined by HPLC analysis, suggesting the purity of up to 98%.
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