The immunofluorescent staining of cells grown on glass slides was performed as described elsewhere [7 (link), 8 (link), 20 (link)]. Cells were fixed with 4% paraformaldehyde, permeabilized with 0.5% Triton X-100 and incubated at 4°C overnight with rabbit polyclonal antibodies against TIN2 [20 (link)], γ-H2AX (i.e., phosphorylated-H2AX) (Upstate), or AR (AR-N20; Santa Cruz), or mouse monoclonal antibodies against AR (AR-414; Santa Cruz), AR-V7 (AG10008; Precision) or pATM (10H11-E12, which detects phosphorylation of ATM at serine 1981; Cell Signaling). Cells were then washed and stained with FITC-labeled goat-anti-rabbit-IgG and/or Texas Red-labeled goat-anti-mouse-IgG (Molecular probes) secondary antibodies [7 (link), 8 (link)]. Images of cells were acquired on an LSM-410 confocal microscope (Zeiss). Labeled foci were counted in enlarged photographs.
Fitc labeled goat anti rabbit igg
Manufactured by Thermo Fisher Scientific
Sourced in United States
FITC-labeled goat-anti-rabbit-IgG is an antibody conjugate designed for detection and visualization applications. It consists of a goat-derived antibody specific to rabbit immunoglobulin G (IgG) that is covalently labeled with the fluorescent dye fluorescein isothiocyanate (FITC). This product can be used to identify the presence and location of rabbit IgG in various experimental procedures.
Automatically generated - may contain errors
Lab products found in correlation
Lsm 410 confocal microscope, by Zeiss (1 mentions)
Ar n 20, by Santa Cruz Biotechnology (1 mentions)
Fluorescence microscopy, by Zeiss (1 mentions)
Triton x 100, by Merck Group (1 mentions)
Prolong gold antifade reagent, by Thermo Fisher Scientific (1 mentions)
Bx53 fluorescence microscope, by Olympus (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
Glutaraldehyde, by Merck Group (1 mentions)
Lps from e coli serotype o55 b5, by Merck Group (1 mentions)
Antibodies against inos and cox 2, by BD (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Phospho jnk, by Cell Signaling Technology (1 mentions)
Goat anti mouse igg, by Thermo Fisher Scientific (1 mentions)
Nf κb, by Cell Signaling Technology (1 mentions)
4 protocols using fitc labeled goat anti rabbit igg
1
Immunofluorescent Staining of Cells
2
Viral Antigen Identification via Immunofluorescence
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The fixed cells were permeabilized with Triton X-100, 0.1% (Sigma–Aldrich GmbH, Steinheim, Germany). For the identification of virus-infected cells, the viral antigens were incubated with hyperimmune serum against ILTV and IBV containing 10% normal goat serum at 37 °C for 1 h, followed by washing with PBS for 3 times. Subsequently, the cells were incubation with fluorescein isothiocyanate (FITC)-labeled goat anti-rabbit IgG (Molecular Probes) for 1 h at 37 °C followed by washing for 3 times with PBS. Finally, the stained cells analyzed by fluorescence microscopy (Carl Zeiss, Germany) [58 (link)].
3
Immunofluorescence Staining of Plasmodium Stages
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Plasmodium berghei schizonts, gametocytes, and subsequent developmental stages (ookinetes and sporozoites) were washed once in PBS and then fixed with 4% paraformaldehyde (Sigma, CA, United States) and 0.0075% glutaraldehyde (Sigma) in PBS for 20 min at room temperature. After washing in PBS, parasites were rinsed with 50 mM glycine in PBS and blocked with PBS containing 5% defatted milk for 1 h at 37°C. After three subsequent washes in PBS, the slides were incubated with an anti-TatD-like DNase serum (1:500 dilution) diluted in 5% defatted milk at 37°C for 1 h and then incubated with FITC-labeled goat anti-rabbit IgG (1:500, Invitrogen, CA, United States) at 37°C for 1 h. The parasite nuclei were counterstained with 1 μg/ml DAPI. ProLong® Gold Antifade Reagent (Invitrogen) was added to the samples and images were captured with an Olympus BX53 fluorescence microscope (Olympus Corporation).
4
Assessing Inflammatory Pathway Modulation
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Pyrroloquinoline quinine (PQQ) disodium salt was purchased from Wako (Wako Pure Chemical Industries, Ltd. Osaka, Japan). Dulbecco's Eagle's Medium (DMEM), fetal bovine serum (FBS), penicillin and streptomycin, and trypsin/EDTA were purchased from Gibco. LPS from E.coli serotype O55:B5 was from Sigma–Aldrich (St. Louis, USA). Antibodies against p38, JNK, phospho-p38, phospho-JNK and NF-κB were purchased from Cell Signaling Biotechnology (Hertfordshire, England). Antibodies against iNOS and COX-2 were from BD Biosciences (Laguna Hills, CA, U.S.A.). Antibody against β-actin and the secondary antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, U.S.A.). FITC labeled goat anti-rabbit IgG and goat anti-mouse IgG were purchased from Invitrogen (Frederick, MD, USA). Enhanced Chemilumincescence (ECL) kit was from Millipore (Amersham Pharmacia Biotech, Piscataway, NJ). PCR primers were synthesized at Invitrogen (Frederick, MD, USA). The total nitrate assay kit was obtained from Beyotime (Nanjing, China)
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