Immuno Staining Techniques
Immunohistochemistry(IHC) & Immunofluorescence(IFA)
Premier Laboratory knows that accurate interpretation of complex microscopic assays begins with excellent sample preparation and staining. Their carefully executed SOPs and personal attention to each and every slide ensures your satisfaction.
Your team at Premier Laboratory has extensive experience developing assays for immuno staining and analysis. Sample preparation, calibration, staining optimization, interpretation and reporting formats may be ordered in a standard or custom format. They have an extensive in-house library of available antibodies, each of which has been validated and optimized for immunohistochemical or immunofluorescent staining. Alternatively, your antibody and/or protocol can be incorporated. The high-throughput automated staining systems at Premier Laboratory result in procedures that are executed with unmatched precision and consistency.
Advantages of immuno and molecular staining:
Microscopic-based immuno staining techniques possess the advantage of being performed on intact tissue sections. Unlike techniques that rely on homogenized or disaggregated samples, the staining results from slide-based techniques are interpreted in context of tissue architecture, morphology and cell type. For complex tissues and heterogeneous samples (eg. breast carcinoma) using morphology as a gating parameter becomes a critical factor in confirming that your results are based on the specific cell population of interest, and therefore valid.
For cell types that cannot be distinguished morphologically, antibodies can be used to locate or exclude an individual population. Multiparametric analysis, with double labeling or antibody cocktails, can be used to further elucidate specific populations.
Scalable resources give you the capacity and precise tools you need for your immuno and molecular analysis. The extensive resources Premier Laboratory will put at your disposal include:
- Pathologists who are board-certified
- A dedicated staff of highly qualified experienced scientists, technologists and technicians
- The very latest in cutting-edge technology including high-throughput automated stainers and image analysis systems
Techniques:
- Immunohistochemistry (IHC)
- Immunofluorescence (IFA)
- New antibody protocol optimization & validation
- Custom antibody panels
- Double/multiple staining protocols
- High-throughput screening with image analysis
- Custom algorithm development (can incorporate automated pattern recognition)
Study design:
Premier Laboratory provides you with consultative and expert advice on a wide array of routine and esoteric research protocols. Each study begins with a consultation to define your study objectives, allocate resources, and determine the optimal scientific approach. When combined these result in the design of the most cost effective, accelerated and efficacious study to meet your research goals. Their scalable product offering means you will achieve precisely the right combination of standard off-the-shelf and custom methodologies in the design of your research study. SOPs and systems are in place to support validated regulated (GLP) and non-regulated investigative protocols.
Studies can be customized to include multiple antibodies, as well as to specify the level of interpretation. The scientific team at Premier Laboratory is always available to develop new protocols tailored to your exact needs.
Antibody selection:
Select from Premier Laboratory’s extensive in-house library of high-quality antibodies that target known protein targets. Most antibodies have been characterized, validated and optimized for immunohistochemical and immunofluorescent staining, with standard protocols in place. Alternatively, your own antibody can be optimized for staining, or a Premier Laboratory scientist will assist you with the selection of the most appropriate commercially available antibody for your study.
Premier Laboratory has extensive experience with the selection of single antibodies or antibody cocktails to provide co-localization information, as well as in double labeling. Your consulting scientist can assist you in selecting the right markers for differentiating the specific cell types needed for your research study.
Antibody optimization and antigen retrieval:
Antibodies are optimized for strong signal-to-noise ratio, thereby eliminating confounding data that can result from nonspecific endogenous background staining. In addition, antigen retrieval techniques may be implemented to unmask binding sites that became unavailable during sample fixation and processing. These techniques allow for maximizing assay sensitivity and specificity, providing you with the most reliable data.
When selecting an antibody from Premier Laboratory’s extensive in-house library, each has already been characterized, validated and optimized for immunohistochemical and immunofluorescent staining, with standard protocols in place.
Calibration and control standards:
All Premier Laboratory equipment is fully validated and calibrated on a scheduled basis. Antibody-based protocols incorporate both positive and negative controls. Control selection is carefully matched to the individual protocol, and is determined during your study design consultation.
Protocols:
Premier Laboratory protocols are developed with the right combination of standard off-the-shelf and custom methodologies to help you reach your research goals. Alternatively, they can implement your own study protocol. During your study design consultation, a Premier Laboratory’s scientist will work with you on the best choice of:
- Antibodies
- Sample processing technique
- Selection of chromagen or fluorochrome and counterstain
- Need for signal amplification
- Staining procedure
- Level of analysis/interpretation
- Number of samples for significant statistical analysis
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