What is IHC and it’s detection methods?

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What is IHC and its Detection Methods?
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Introduction
Immunohistochemistry (IHC) refers to the course
of action of becoming aware of antigens like
proteins in cells of a tissue section taken
during biopsy. It is a molecular test that
involves the make use of antibodies to spot
specific proteins within tissues on microscope
slides.  IHC is extensively utilized for
diagnosis of specific tumor antigens.  
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IHC Detection Methods
There are two IHC detection methods. These are
direct and indirect detection which are used to
fluorescents and chromogenic signal for protein
detection.  In direct detection, the main
antibody is exact for the target molecule is
straightforwardly labeled. Indirect detection
makes use of an unconjugated primary antibody. As
an alternative, a secondary antibody raised
against the species of the primary antibody
carries the label. The indirect method is
frequently the favored technique in
immunohistochemistry (IHC) staining experiments
because of signal amplification courtesy of
secondary antibodies. 
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Direct Detection
The direct detection of IHC detection method has
been achieving reputation, particularly in the
midst of researchers doing immunofluorescence and
flow cytometry studies, for a number of causes.
For one, direct detection eliminates the need for
an additional incubation step and a secondary
reagent. The primary antibody can be unswervingly
labeled and conjugated to an appropriate enzyme
(e.g., horseradish peroxidase (HRP), alkaline
phosphatase (AP)) or fluorochrome. In adding
together, the accessibility of a vast range of
fluorochromes makes designing multicolor testing
a breeze.
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• In spite of the obvious advantages, there are
  some disadvantages in utilizing the direct
  detection method
• It is not as sensitive as the indirect detection
  methods. The direct detection technique is
  idyllic for applications where there is an large
  quantity of target proteins and the primary
  antibody is extremely specific.
• The primary antibody is tagged with a label,
  making it inappropriate for other applications
  down the line.
• Making an allowance for the fact that there is a
  inadequate range of directly labeled primary
  antibodies available, you may require to label
  your own antibodies.

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Indirect Detection
There are a number of types of IHC detection
methods of indirect detection. The most accepted
and possibly most established one is probably the
ABC method. However, polymer based detection kits
that function in a similar manner are becoming
increasingly popular. Kits that are capable of
detecting several species of primary antibody
while amplifying signal are particularly
versatile. Chromogenic detection has the benefit
of being extremely stable and not being
vulnerable to photo-bleaching. When performed in
paraffin entrenched tissue sections the slides
are very tough and can be examined by several
people without having to take pictures right away
or worry about photo-bleaching.
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Signal intensification techniques such as the ABC
more than overcompensate for any benefit in
sensitivity that a straightforwardly labeled
fluorescent derivative antibody would provide.
This is more often than not the method of choice
for analyzing paraffin embedded tissue sections.
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