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LifeSpan's Immunohistochemistry Services
[Full Service Protein Localization]
[The Utility of Molecular Pathology] |
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| LifeSpan's immunohistochemistry services provide high-quality
information about the localization of proteins in normal and pathologic
tissues. For each protein studied, optimal antigen retrieval and IHC labeling
conditions are identified, tissues are immunostained and analyzed by an
anatomic pathologist, and results are presented through an online client
interface. All custom protein expression-profiling studies are confidential and
are performed on a fee-for-service basis without royalties or milestone
payments.
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| Skin, Epidermis |
Brain, Purkinje Neuron |
Pancreas, Islet (double label) |
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| Full Service Protein
Localization
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| LifeSpan's comprehensive services allow customers to outsource their entire
localization project, from biological question to final report, without having
to worry about any of the logistics. Standard services include: |
| Study Design:
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Each contract research study begins with a customer
consultation to define the study goals, available resources, and the optimal
scientific approach. LifeSpan has conducted thousands of localization studies
for more than 100 companies worldwide, experience that can be applied to help
ensure a successful study.
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| Antibodies/QC:
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Antibodies can be provided by the client, purchased
from a commercial resource, acquired from LifeSpan's
antibody inventory or
custom generated by LifeSpan.
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| Tissues/QC:
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Tissue specimens are selected that will best answer
the biological question. Normal and diseased, frozen and formalin-fixed, human
and non-human tissues can be supplied by the client, acquired from LifeSpan's
extensive
tissue bank, selected from LifeSpan's
inventory of
tissue microarrays, purchased from
commercial vendors, or collected prospectively by LifeSpan for use in the
study. LifeSpan also offers custom tissue microarray design. Prior to use in
any study, all tissues are subjected to rigorous testing to ensure proper
fixation, accurate diagnosis, and antigen validation.
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| Assay Optimization:
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Antibodies are initially tested in Phase I to
ensure that they are working appropriately before being used to screen the
selected test tissues in Phase II. This involves performing a serial
dilution of the antibody to establish the optimal staining concentration.
Optimizing antigen-retrieval conditions may also be necessary to improve the
antibody's signal-to-noise ratio on tissue samples.
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| Immunohistochemistry:
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During Phase II testing, antibodies are used under the
optimized assay conditions to immunolabel the selected tissues of interest.
LifeSpan can use single or multiple antibodies to provide co-localization
information. LifeSpan has extensive experience in double labeling and can
recommend markers for differentiating specific cell types.
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| Interpretation:
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The resulting immunolabeled slides are interpreted by
LifeSpan's staff of pathologists, who have many years experience interpreting
the often complex behavior of antibodies in IHC. The cellular localization
profile is then correlated with the normal and/or disease processes being
evaluated.
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| Quantitative Analysis:
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In addition to standard pathology scoring (subjective
analysis using a scale of 0-4), studies can also be expanded to include
quantitative analysis. Such analyses are customized for each study and include
manual masking of cell types or regions for analysis, and automated signal
quantification of single or multiple white light chromogens (i.e. DAB, Vector
Red).
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| Detailed Reporting:
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LifeSpan has a flexible report format that allows for customization depending
upon the level of interpretation required. The two most common forms are Full
Text and Tabular reporting.
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| Example of Full Text Reporting |
Example of Tabular Reporting |
Full-Text Reporting: With Full-Text Reporting, LifeSpan's pathologists
describe the immunolocalization patterns in detail and provide 3-5
representative images for each tissue sample. This format is useful for studies
that require detailed observations and interpretation, such as studies
involving CNS tissues, co-localization studies, or those that may involve rare
events and minor cell types. Information about each patient sample is also
provided.
Tabular Reporting with and without Imaging: With Tabular Reporting,
LifeSpan's pathologists quantify the signal present in major cell types within
each tissue sample in a 0-4 scale and present the results in a table. This
format is useful for studies at the initial stages of target characterization,
screening studies, or those involving major cell types or in homogeneous
tissues. Customers can choose to include selected images for each tissue
sample.
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| Study Deliverables:
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Upon the conclusion of each study, LifeSpan will
provide a report detailing the assay condition, the antibody dilution
specifications, and localization results for each tissue specimen assayed.
Selected patient information, including the age, sex, cause of death or
surgery, is provided with each sample. LifeSpan offers a variety of reporting
formats to meet the varied needs of our customers. Reports are delivered via
secure website and can also be delivered on CD.
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| Timeline:
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Standard IHC studies take from 3-12 weeks to complete,
depending upon the complexity of the study. LifeSpan strives to complete our
customers' work in an expeditious manner.
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| Cost:
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LifeSpan offers unparalleled quality and competitive
pricing for its contract research services. Due to the variable nature of these
studies, a custom price is quoted for each study.
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| The Utility of
Molecular Pathology
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| Knowing which cell types express a protein and how those cell types behave in
normal and disease processes is of great value during the drug discovery and
development process. The resources needed to gain this understanding are often
difficult to acquire, such as immunohistochemistry-validated antibodies,
quality normal and diseased human tissues, and the experience and ability to
optimize assay conditions, interpret real signal from background, and correlate
expressing cell types with human pathologies. LifeSpan has been providing these
resources to companies worldwide for nearly a decade in the form of proprietary
reagents, contract research services, and human protein localization reports.
Frequent applications of molecular pathology are listed below. |
| Supporting RNA data with protein immunohistochemistry |
| Immunohistochemistry is an important method for validating observations made by
using nucleic acid based expression analysis, such as RT-PCR or gene-chip array
technologies. It is important to determine whether observed changes in mRNA can
also be seen in the translated protein, and to understand what cell types are
exhibiting these changes. Additional mRNA copies may be transcribed but never
translated into protein while increased expression may be the result of
up-regulation in native cells or due to an increase in infiltrating
inflammatory cells. |
| Differentiating discrete cell types and co-localization |
| Because whole tissues are used in IHC rather than homogenized samples, a
pathologist can differentiate between the many different cell types that make
up a complex tissue. In cases where cell types can not be distinguished from
one another morphologically, such as the pancreatic alpha and beta cells,
LifeSpan can double-label with cell specific markers to provide co-localization
information. |
| Understanding prevalence within a disease |
| With the emergence of commercial tissue microarrays (TMAs), it is now possible
to quickly and cost-effectively use IHC to study a target protein's prevalence
within a larger population of samples. TMAs are appropriate for answering
specific biological questions and LifeSpan has the expertise to validate them
for fixation, pathology, and antigenicity before they are used in a study. |
| Investigating expression during disease progression |
| Understanding a protein’s role during disease progression is important when
defining its potential as a therapeutic target. Changes in expression may occur
upon disease onset, persist throughout progression, or only occur at specific
stages. This information can be used to define applications for therapeutics
and improve efficiencies of the development process. LifeSpan can provide
multiple disease grades and stages for inclusion in progression studies. |
| Understanding potential toxicology issues |
| Understanding a protein's histological expression profile can provide valuable
information about the potential toxicologic effects of targeting it with a
therapeutic agent. Evidence such as neuronal staining could influence how an
antagonist molecule is formulated. Observed staining in replenished cell types,
such as gastro-intestinal epithelium can help predict side effects upon
treatment. Localization to critical cell types, such as hepatocytes or bone
marrow precursors, may de-prioritize one target in favor of another with a more
restricted expression and localization profile. |
| Understanding a protein target's normal tissue expression
profile |
| A protein's normal expression profile can be understood by using
immunohistochemistry to screen broad panels of normal tissues. The resulting
cell-specific staining patterns can then be used to predict a protein's
involvement in normal and diseased biological processes. The LifeSpan
DrugTargetTM Database contains whole body expression profiles for
hundreds of proteins while normal human tissue microarrays are available for
use in profiling additional targets of interest. |
| Understanding expression in complex tissues |
| Subtle differences in localization can have significant meaning when studying
expression in complex tissues, such as the human hypothalamus, rat brain, and
the developing fetus. LifeSpan's staff of human and veterinary pathologists
have the expertise needed to map these tissues and analyze the subsequent
immunolabeling. |
| Searching for new disease indications |
| Using IHC to screen broad panels of diseased tissue types can be useful in the
identification of alternative disease indications for therapeutic intervention.
Understanding a protein's histological profile in normal tissues and
correlating the observed expressing cell types with function can also be useful
in the identification of diseases in which a drug target may play a role. Each
LifeSpan Gene Report contains immunohistochemical profiles for extensive panels
of normal and diseased tissues. |
| Understanding complex expression patterns within disease |
| Cancer is a disease that commonly exhibits heterogeneous expression due to
subpopulations of rapidly proliferating tumor clones as well as changes in
vascular proliferation (e.g., angiogenesis), desmoplasia, and accompanying
inflammatory responses. Immunohistochemistry coupled with multiple full size
cancer specimens of various stages is useful in studying these expression
variations. |
| Selection of the most appropriate animal models |
| In an effort to provide the most cost-effective, timely services possible,
LifeSpan maintains a pre-validated inventory of animal tissues that are
commonly requested for use in contract research studies. These tissues are
reviewed by a pathologist for fixation quality and diagnostic accuracy, and
each sample is tested with control antibodies to ensure antigen preservation
prior to being used in a study. |
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