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Western Blot – Multimedia

Comparison of Key Signaling Pathways in Lung Cancer Cells Grown in 2D and 3D Formats content piece image
Poster

Comparison of Key Signaling Pathways in Lung Cancer Cells Grown in 2D and 3D Formats

Evaluation of Wnt/ß-catenin and BMP pathway proteins and gene expression in NSCLC cell lines A549 and H460 in 2D and 3D cultures.
App Note / Case Study

Validating the Expression Consistency of a Housekeeping Protein

Housekeeping proteins (HKP) are typically used as loading controls because it is assumed that the HKP expression level remains consistent across samples. However, there is evidence that HKP expression levels change in many scenarios, including siRNA treatment, cell death, cell differentiation, etc.
App Note / Case Study

A Method for Greater Reliability in Western Blot Loading Controls: Stain-Free Total Protein Quantitation

Reliable assessment of changes in target protein levels by western blot requires measurement of both the target and loading control proteins in the linear dynamic range. Stain-free technology is a novel method introduced by Bio-Rad to visualize and quantify proteins in gels and blots.
App Note / Case Study

General V3 Western Blotting Protocol

Western blotting is a very useful and widely adopted lab technique, but the traditional procedure can be long and tedious. Researchers can assess only whether a blot image is captured at the end of the long procedure and the quality of western blot data is sometimes challenged due to poor loading controls.
App Note / Case Study

Determining the Appropriate Sample Load When Using a Stain-Free V3 Western Workflow™

Traditional western blot workflows require an appropriate sample load to detect both target proteins and loading control proteins. The discrepancy between these quantities often leads to oversaturated protein bands, forcing researchers to sometimes run two duplicate gels or titrate down the antibody concentration in order to get quantitative results.
App Note / Case Study

Determining the Appropriate Sample Load for Western Blots

Reliable western blot data can be generated only when the proper sample amount of protein is used. Loading too much protein leads to signal saturation in western blots, yet too little produces weak signals.
App Note / Case Study

Avoiding Housekeeping Protein Detection Saturation

Reliable western blot data require detection of the target and loading control proteins in the linear dynamic range. Many published western blot images show saturated protein bands, indicating that they were not measured in the linear dynamic range. Quantitative analysis based on this type of data is not reliable.
App Note / Case Study

BD Cytometric Bead Array Cell Signaling Flex Set System

The BD™ Cytometric Bead Array (CBA) Cell Signaling Flex Set system lets you achieve high productivity in cell signaling studies by extracting more data from less sample faster than traditional Western blot techniques.
Intended transcriptional gene silencing with siRNA to the human Vascular Endothelial Growth Factor (VEGF) promoter results in VEGF repression through sequence-specific off-targeting content piece image
Poster

Intended transcriptional gene silencing with siRNA to the human Vascular Endothelial Growth Factor (VEGF) promoter results in VEGF repression through sequence-specific off-targeting

SiRNA designed to target the human VEGF gene promoter revealed a putative candidate for transcriptional gene silencing. However, mutation or deletion of the target site demonstrated that the observed VEGF knockdown was the result of a sequence-specific off-target effect. Bioinformatic and microarray analyses of genes implicated in VEGF transcriptional control followed by knockdown and Western blotting experiments presented one candidate, GRB2, as an unintended target of the VEGF promoter-specifi
LacZ Reporter Fusion Assay for Rapid and Easy Identification of Highly Efficient siRNA and its Delivery by Lentivirus into Suspension Cell Lines content piece image
Poster

LacZ Reporter Fusion Assay for Rapid and Easy Identification of Highly Efficient siRNA and its Delivery by Lentivirus into Suspension Cell Lines

The LacZ reporter fusion assay is an excellent method to precisely quantify knockdown efficiency of siRNA-sequences and can be corroborated by Western blot analysis. Efficient production of Lentiviruses and high transduction efficiency (up to 98%) were achieved on lymphoid B- and T-cells as demonstrated by FACS analysis and GFP expression. Successful knockdown effect with specific shRNA was observed in suspension cells three days after lentiviral infection.
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