|
1. Explanation for immunohistochemistry service |
All of the Tissue Array slides are collected and arranged by pathologists.
The tissues were fixed in 4% buffered neutral formalin for 12-24 hours,
dehydrated by gradient ethanol, cleared by xylene and embedded in
paraffin. All of the tissues are tested for immunohistochemistry on
several antibodies including cytokeratin and vimentin. Some of the
paraffin blocks are 10 or more years old. However, the staining quality
is not different from the tissue of new paraffin blocks.
If requested, TEST IMMUNOHISTOCHEMISTRY STAINING is performed to determine
whether your antibody is useful for immunohistochemistry. Several
antigen retrieval methods are added if necessary. For this test, formalin
fixed tissues of normal organs are used. We perform the immunohistochemistry
on tissue array slides if the above test yields satisfactory result.
Test Immunohistochemistry is necessary to obtain the optimal staining
condition.
Tissue array slide is the easiest and most effective technique for
analyzing the Expressional Profile Analysis. You can either purchase
the Tissue Array slides or order immunohistochemistry service. If
you are not confident with the microscopic interpretation, our experienced
pathologists can analyze the expression profile. In that case, you
will receive the Expression Profile Analysis Sheet with a summary
statement, and stained tissue array slides along with digital images
of most tissues.
For humanized antibody, biotinylation is followed by immunohistochemistry.
The immunohistochemistry service usually takes 3 weeks including Test
Immunohistochemistry and Expressional Profile Analysis. |
 |
|
|
2. Protocol for immunohistochemistry |
Depaffinization and hydration
- incubate in a dry oven at 60 degree for 1 hour
- dewax slides in xylene for 4 minutes x 5 times
- hydrate slides in 100%, 95% and 75% ethanol for 3 minutes x
2 times each.
- immerse slides in tap water for 5 minutes
Antigen retrieval method (optional)
- immerse slides into citrate buffer (0.01 M, pH 6.0)
- microwave (700 W or high) for 5 min (add citrate buffer if necessary
)
- microwave (medium) for 5 min (add citrate buffer if necessary)
- microwave (low) for 5 min, then immerse in cold PBS
Quenching endogenous peroxidase (optional)
- immerse slides in 3% hydrogen peroxide solution for 6 minutes
- wash slides in PBS for 5 minutes x 3 times
Primary antibody
- incubate slides with blocking serum for 30 min (*)
- blot excess serum from sections
- incubate with diluted primary antibody (**)
- wash slides in PBS for 5 minutes x 3 times
Secondary antibody
- incubate slides with biotin-conjugated secondary antibody for
30 min (*)
- wash slides in PBS for 5 minutes x 3 times
Avidin-biotin-peroxidase complex
- incubate slides with Avidin-Biotin Complex for 30 min (*)
- wash slides in PBS for 5 minutes x 3 times
Chromogenic reaction
- incubate slides in fresh DAB solution for 2 minutes (***)
- stop the reaction by washing in tap water
Counterstain
- in Mayer's hematoxylin for 10 seconds
Dehydration, clearing and mount
- dehydrate slides in 75%, 80%, 95% and 100% ethanol for 1 min
each
- clear slides in xylene 5 minutes x 4 times
- mount the cover slide with Permount
* Blocking serum, secondary antibody and avidin-biotin-peroxidase
complex are included in most of the immunostaining kit. Our Lab use
the ABC kit from Vector Lab (Vestastain Elite ABC kit) PK-6102 for
mouse antibody and PK-6101 for rabbit antibody
** Incubation time varies according to antibodies. The followings
are recommended as the first trial, when you do not know the optimal
condition.
- Monoclonal antibody; 2 hours at room temperature or overnight
at 4 degree
- Polyclonal antibody; 1 ~ 1.5 hours at room temperature
*** We use DAB solution in Vector DAB/Ni substrate kit (Cat. SK-4100)
|
|
|
|
3. Price for Immunohistochemistry Service |
PACKAGE
SA0. Expression analysis of a antibody on human major normal tissues:
$1300. The price includes (1) evaluation of antibody, (2) staining, (3) interpretation
and (4) photographs of major human organs)
INDIVIDUAL ITEMS
SA1. Evaluation and adjustment of optimal staining conditions: $450
per antibody. The price includes the slides.
SA2. Immunohistochemistry: $120.00 per slide (The price does not includes
the cost of slides or reading fee.)
SA3. Expressional profile analysis on human major normal tissues:
$750 per antibody. (Interpretation of immunohistochemistry slide by experienced pathologists.
Price does not include the slide cost or immunohistochemistry fee.
Price includes analysis sheet and microphotographs of most organs)
SA4. Expressional analysis on tumor tissues: $250 per slide. (Interpretation
of immunohistochemistry slide by experienced pathologists. Price does
not include the slide cost or immunohistochemistry fee. Price includes
interpretation result and microphotographs of most organs)
SA5. Expressional profile analysis on rat or mice major normal organ:
$450.00 per antibodies. (Interpretation of immunohistochemistry slide by experienced pathologists.
Price does not include the cost of slides or immunohistochemistry
fee. Price includes analysis sheet and microphotographs of most organs) |
|
|
|
4. Expressional profile analysis sheet |
|
|
|
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;){kind=small}
;)