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Immunofluorescence in Biological Technology

Sigma-Aldrich and Atlas Antibodies Announce 2,300 New, Highly Validated Prestige Antibodies(RM)

... Prestige Antibody is also associated with over 500 tissue immunohistochemistry (IHC) images, over 100 cell/line immunocytochemistry (ICC) images, immunofluorescence (IF) and western blotting (WB) data, all publicly accessible via the Human Protein Atlas ( www.proteinatlas.org ). In August 2008, ...

Proteomics in Bladder Cancer Research: Protein Profiling of Bladder Squamous Cell Carcinomas, Rev C

... about protein expression profiles and immunofluorescence patterns that may be associated with ... is indicated with small arrows, showed immunofluorescence patterns that were clearly different ... we show in Figures 4A, D, G, J, and M immunofluorescence pictures of cryostat sections from ...

Visualizing siRNA in Mammalian Cells: Fluorescence Analysis of the RNAi Effect

... 48 hr post-transfection using immunofluorescence microscopy (Figure 1A). The results clearly ... were analyzed for c-myc protein expression using immunofluorescence as described in Materials and Methods. ... in only 2 or 3 days. Supporting immunofluorescence data showed that the siRNA was ...

Drive Long-Term Silencing with CMV Vectors

... to non-transfected HeLa cells. Panel B shows immunofluorescence data demonstrating the relative ... clones using either real-time PCR or immunofluorescence analysis. (A) Clones that showed low ... amount of GAPDH knocked down. immunofluorescence was performed using gene specific ...

Dicer vs. RNase III for Preparation of siRNA Cocktails

... GAPDH protein levels were monitored by immunofluorescence 48 hours after transfection ( B ) and ... the reduction in protein levels was examined by immunofluorescence microscopy. Fluorescent signal was ... RNase III generated siRNAs to GAPDH. immunofluorescence analysis of GAPDH, La, c-MYC, Cdk-2, Ku-90, and ...

Inducing RNAi with siRNA Cocktails Generated by RNase III

... population was transfected into 293 cells. immunofluorescence was used to examine the gene silencing ... at 48 hours post transfection and immunofluorescence was performed with the appropriate ... levels. Figure 2B shows representative immunofluorescence images demonstrating reduction in ...

GENE SILENCING WITH PCR PRODUCTS

... assessed using nuclear staining with DAPI and immunofluorescence using a c-FOS antibody detected with a ... using nuclear staining with DAPI and immunofluorescence using a GAPDH-specific antibody. The DAPI and immunofluorescence images are overlaid in the figures. ...

Five Ways to Produce siRNAs

... protocol, at a final siRNA concentration of 75 nM. immunofluorescence analysis was performed 96 hr post transfection using mouse ... the cells were assessed using nuclear staining with DAPI and immunofluorescence using a c-FOS antibody. Non-transfected cells (NT) as well as ...

Control Your siRNA Research

... Each of these primary antibodies has been used in immunofluorescence experiments at Ambion to detect the reduction in protein levels ... in solution in a 100 g unit size and are validated for use in immunofluorescence experiments. Note that the concentration and volume will vary ...

Cancer-Related miRNAs Uncovered by the mirVana miRNA Microarray Platform

... NeoFX Transfection Agent (Ambion). immunofluorescence assays were used to measure the two putative oncogene targets ... introduced into cells that express high levels of the miRNA. immunofluorescence 48 hours after transfection revealed an approximately two-fold ...

Efficient RNAi-mediated gene silencing in neuronal cells using QIAGEN siRNA and TransMessenger Transfection Reagent*

... Cells were stained and analyzed after transfection. Double immunofluorescence labeling and image analysis was performed as described previously ... was. Cells were targeted with siRNA 58 days after plating. Double immunofluorescence labeling of MAP2 and the unrelated protein actin was performed 48 ...

Efficient DNA transfection of primary CNS neurons using TransMessenger Transfection Reagent

... were fixed using 4% paraformaldehyde 24 hours after transfection. immunofluorescence was performed according to standard procedures using primary ... cultures display the same GFAP- and MAP2-staining pattern. The immunofluorescence results show that primary cultures of cortical neurons generated ...

Mix-and-Read, Bead- and Cell-Based Immunoassays Using the FMAT 8100 HTS System

... ligand binding4, apoptosis5, immunofluorescence5, and bead-based fluorescent immunoassays (FLISAs) 3. In the experiments discussed here, immunofluorescence and FLISA assays were combined in a single well, and the secreted cytokines and cell surface expression of an inducible marker were quantitated ...

Fluorescently Labeling Long dsRNA with the Silencer siRNA Labeling Kit

... control protein, Pab 1, was not affected (Figure 1A). The specific silencing of hrp48 and U2AF 50 proteins was also confirmed by immunofluorescence microscopy (data not shown). Direct observation of the Cy3 fluorescence demonstrated that both hrp48 and U2AF 50 labeled dsRNA ...

Using Validated siRNAs in Functional Genomic Assays

... they were transfected and transferred to wells of a 96 well plate. Following overnight recovery, the cells were processed for immunofluorescence staining of NF-kappaB. There are several distinct signaling pathways that can trigger the nuclear translocation of NF-kappaB. ...

The let-7 microRNA Family Regulates RAS: Implications for Development and Oncogenesis

... Inhibitors (Ambion) or a negative control miRNA inhibitor (CD). Three days post-transfection, RAS expression was monitored by immunofluorescence (A, C) and quantified using MetaMorph software (Universal Imaging corporation) (B, D; triplicate assays). Modified with ...

Superior Gene Silencing Using Adenoviral Vectors

... to the wild type construct, whereas that of Ad-C10i was complementary to the mutant construct. (A) Green: GFP. Red: immunofluorescence of the polyQ construct detected with 1C2 antibody. Ad-G10i reduced wt ataxin-3 levels 95% but did not affect mutant ataxin-3 ...

RNAi: A How To for New Users

... its effects can be easily monitored by qRT-PCR or other methods at the mRNA level, or by Western blot or immunofluorescence at the protein level. In fact, Ambion provides a highly specific antibody for measuring these protein level ...

RNAi in Non-Mammalian Cultured Cells

... Pab 1, was not affected (Figure 3A). The specific silencing of hrp48 and U2AF 50 proteins was also confirmed by immunofluorescence microscopy (data not shown). Direct observation of the Cy3 fluorescence demonstrated that both hrp48 and U2AF 50 ...

More siRNA Vectors for RNA Interference

... than the mRNA-specific probes. (B) GAPDH protein levels in cells transfected with p Silencer 2.0-U6-GAPDH were analyzed by immunofluorescence using a GAPDH-specific antibody ....

Matched siRNAs and Assays: Ambion + Applied Biosystems = RNAi Success

... reduced survivin mRNA levels in these cells by 80% compared to cells treated with the negative control siRNA (Figure 5). In addition, immunofluorescence analysis of survivin protein levels indicated that protein levels were reduced 76% compared to cells transfected with the ...

Delivering siRNA Using Adenoviral Vectors

... or a scrambled negative control siRNA. The media was changed 4 hours after infection and cells were analyzed after 72 hours. immunofluorescence was performed using mouse anti-GAPDH antibody. Green: GAPDH protein. Blue: DAPI stained nuclei. ...

Choose the Right Transfection Agent for Your RNAi Experiment

... Kit. After transfection of the positive control, reduction in GAPDH activity can be monitored by immunofluorescence or Western blot with Ambion's monoclonal antibody to GAPDH, or by Northern blot, RT-PCR or RPA. ...

Avoid siRNA Design Altogether!

... synthesized siRNA known to efficiently silence GAPDH, was transfected into HeLa cells. GAPDH protein levels were analyzed by immunofluorescence 48 hrs later. back to top Ordering Information ...

Assessing Gene Function with siRNA Libraries

... phenotypes were then examined by microscopic readout to determine their root causes. This was done by immunofluorescence staining of multiple markers, including tubulin to reveal microtubule distribution. This approach revealed ...

Sensitive detection of tumor cells in peripheral blood of carcinoma patients by a reverse transcription PCR method

... diagnostic approach is designed to optimize enrichment and analysis of tumor cells. The specificity of the method is shown by immunofluorescence staining (figures 1 and 2) and the lower detection limit by RT-PCR (figure 3). Specificity Figure 1 shows ...

Detection of mRNAs on Cryosections of the Cardiovascular System Using DIG-Labeled RNA Probes

... were briefly rinsed in distilled water, then counterstained with methylene green. The sections were mounted with Kaisers glycerin-gelatin. If immunofluorescence protocols are used in combination with in situ hybridization, we recommend development with the ELF substrate (ELF Kit, Molecular Probes). ...

Effectene Transfection Reagent provides efficient gene delivery to primary neuronal cell cultures

... in triplicate. Control cells were exposed to Effectene Reagent complexed with plasmid DNA lacking a CMV eukaryotic promoter. immunofluorescence was performed as previously described (2). Efficient Transfection Using Effectene Reagent ...
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