Pan Cytokeratin [AE1/AE3]

Pan Cytokeratin [AE1/AE3]
Pan Cytokeratin [AE1/AE3]
Concentrated and Prediluted Antibody Cocktail
901-011-071217
Catalog Number:
CM 011 A, B, C
IPI 011 G10
VP 011 G, G25
Description:
0.1, 0.5, 1.0 ml, concentrated 6.0, 25 ml, prediluted
PM 011 AA, H
10 ml, prediluted
6.0, 25 ml, prediluted
Dilution:
1:100
Ready-to-use
Ready-to-use
Ready-to-use
Diluent:
Da Vinci Green
N/A
N/A
N/A
Protocol Recommendations (intelliPATH and manual use)
Cont'd:
Digestion Method:
Digest with Trypsin enzyme for 5 minutes at 37ºC -or- for 20 minutes
at RT.
Protein Block (Optional): Incubate for 5-10 minutes at RT with
Biocare's Background Punisher.
Primary Antibody: Incubate for 30 minutes at RT.
Probe: Incubate for 10 minutes at RT with a secondary probe.
Polymer: Incubate for 10-20 minutes at RT with a tertiary polymer.
Chromogen: Incubate for 5 minutes at RT with Biocare's DAB -ORIncubate for 5-7 minutes at RT with Biocare's Warp Red.
Counterstain: Counterstain with hematoxylin. Rinse with deionized
water. Apply Tacha's Bluing Solution for 1 minute. Rinse with deionized
water.
intelliPATH™ Automated Slide Stainer:
IPI011 is intended for use on the intelliPATH™ Automated Slide
Stainer. Refer to the intelliPATH Automated Slide Stainer manual for
specific instructions on its use. When using the intelliPATH, peroxide
block with intelliPATH Peroxidase Blocking Reagent (IPB5000) may be
performed following pretreatment.
Protocol Recommendations (Ventana BenchMark ULTRA):
Refer to the User Manual for specific instructions for use.
Recommended protocol parameters are as follows:
Template: U ultraView DAB
Pretreatment Protocol: ULTRA CC1 Standard (64 min) at 95°C
Primary Antibody: Incubate for 32 minutes at 37°C
ultraBlock (V-Blocker BRI4001): Incubate for 4 minutes (with
appropriate Option # registered by user)
V-Blocker is recommended to be applied prior to any detection system.
Detection: ultraView
Technical Note:
1. With cytokeratin markers, heat retrieval may provide a higher
sensitivity assay; whereas, enzyme digestion may produce greater
specificity. Users should validate the pretreatment method for their
specific application.
2. This antibody has been optimized for use with Biocare's MACH 4
Universal HRP-Polymer Detection and intelliPATH Universal HRP
Detection Kit. Use TBS for washing steps.
3. Protocol Recommendation for Ventana BenchMark XT available upon
request.
Limitations:
The optimum antibody dilution and protocols for a specific application
can vary. These include, but are not limited to fixation, heat-retrieval
method, incubation times, tissue section thickness and detection kit
used. Due to the superior sensitivity of these unique reagents, the
recommended incubation times and titers listed are not applicable to
other detection systems, as results may vary. The data sheet
recommendations and protocols are based on exclusive use of Biocare
products. Ultimately, it is the responsibility of the investigator to
determine optimal conditions. The clinical interpretation of any
positiveor negative staining should be evaluated within the context of
clinical presentation, morphology and other histopathological criteria
by a qualified pathologist. The clinical interpretation of any positive or
negative staining should be complemented by morphological studies
Intended Use:
For In Vitro Diagnostic Use
Pan Cytokeratin [AE1/AE3] is a mouse monoclonal antibody cocktail
that is intended for laboratory use in the qualitative identification of a
broad spectrum of acidic and basic cytokeratin proteins by
immunohistochemistry (IHC) in formalin-fixed paraffin-embedded
(FFPE) human tissues. The clinical interpretation of any staining or its
absence should be complemented by morphological studies using
proper controls and should be evaluated within the context of the
patient’s clinical history and other diagnostic tests by a qualified
pathologist.
Summary and Explanation:
AE1/AE3 recognizes the acidic and basic (Type I and II) subfamilies of
cytokeratins. The cocktail of these two antibodies can be used to
detect most human epithelia. The acidic cytokeratins have molecular
weights of 56.5, 55, 51, 50, 50, 48, 46, 45, and 40 kDa. The basic
cytokeratins have molecular weights of 65-67, 64, 59, 58, 56 and 52
kDa. This pan cytokeratin antibody has proved useful as a screener for
the majority of human carcinomas.
Principle of Procedure:
Antigen detection in tissues and cells is a multi-step
immunohistochemical process. The initial step binds the primary
antibody to its specific epitope. A secondary antibody may be applied
to bind the primary antibody, followed by an enzyme labeled polymer;
or an enzyme labeled polymer may be applied directly to bind the
primary antibody. The detection of the bound primary antibody is
evidenced by an enzyme-mediated colorimetric reaction.
Source: Mouse monoclonal
Species Reactivity: Human, mouse and rat
Clone: AE1/AE3
Isotype: IgG1
Total Protein Concentration: ~10 mg/ml. Call for lot specific Ig
concentration.
Epitope/Antigen: AE1/AE3
Cellular Localization: Cytoplasmic
Positive Control: Skin or adenocarcinoma
Known Applications:
Immunohistochemistry (formalin-fixed paraffin-embedded tissues)
Supplied As: Buffer with protein carrier and preservative
Storage and Stability:
Store at 2ºC to 8ºC. Do not use after expiration date printed on vial. If
reagents are stored under conditions other than those specified in the
package insert, they must be verified by the user. Diluted reagents
should be used promptly; any remaining reagent should be stored at
2ºC to 8ºC.
Protocol Recommendations (intelliPATH and manual use):
Peroxide Block: Block for 5 minutes with Biocare's Peroxidazed 1.
Pretreatment Protocol:
Pretreatment may be performed by heat retrieval or enzyme digestion.
Heat Retrieval Method:
Pretreatment: Perform heat retrieval using Biocare’s Reveal Decloaker.
Refer to the Reveal Decloaker product data sheet for specific
instructions.
Rev: 062117
Pan Cytokeratin [AE1/AE3]
Concentrated and Prediluted Antibody Cocktail
901-011-071217
Limitations Cont’d:
using proper positive and negative internal and external controls as
well as other diagnostic tests.
Quality Control:
Refer to CLSI Quality Standards for Design and Implementation of
Immunohistochemistry Assays; Approved Guideline-Second edition
(I/LA28-A2) CLSI Wayne, PA USA (www.clsi.org). 2011
Precautions:
1. This antibody contains less than 0.1% sodium azide. Concentrations
less than 0.1% are not reportable hazardous materials according to
U.S. 29 CFR 1910.1200, OSHA Hazard communication and EC Directive
91/155/EC. Sodium azide (NaN3) used as a preservative is toxic if
ingested. Sodium azide may react with lead and copper plumbing to
form highly explosive metal azides. Upon disposal, flush with large
volumes of water to prevent azide build-up in plumbing. (Center for
Disease Control, 1976, National Institute of Occupational Safety and
Health, 1976) (5)
2. Specimens, before and after fixation, and all materials exposed to
them should be handled as if capable of transmitting infection and
disposed of with proper precautions. Never pipette reagents by mouth
and avoid contacting the skin and mucous membranes with reagents
and specimens. If reagents or specimens come into contact with
sensitive areas, wash with copious amounts of water. (6)
3. Microbial contamination of reagents may result in an increase in
nonspecific staining.
4. Incubation times or temperatures other than those specified may
give erroneous results. The user must validate any such change.
5. Do not use reagent after the expiration date printed on the vial.
6. The SDS is available upon request and is located at
http://biocare.net.
Troubleshooting:
Follow the antibody specific protocol recommendations according to
data sheet provided. If atypical results occur, contact Biocare's
Technical Support at 1-800-542-2002.
References:
1. Bunton TE. The immunocytochemistry of cytokeratin in fish tissues.
Vet Pathol. 1993 Sep; 30(5):418-25.
2. Sorensen SC, et al. Structural distinctions among human breast
epithelial cells revealed by the monoclonal antikeratin antibodies AEI
and AE3. J Pathol. 1987 Oct; 153(2):151-62.
3. Pinkus GS, Etheridge CL, O'Connor EM. Are keratin proteins a better
tumor marker than epithelial membrane antigen? A comparative
immunohistochemical study of various paraffin-embedded neoplasms
using monoclonal and polyclonal antibodies. Am J Clin Pathol. 1986
Mar; 85(3):269-77.
4. Pinkus GS, et al. Optimal immunoreactivity of keratin proteins in
formalin-fixed, paraffin-embedded tissue requires preliminary
trypsinization. An immunoperoxidase study of various tumours using
polyclonal and monoclonal antibodies. J Histochem Cytochem. 1985
May; 33(5):465-73.
5. Center for Disease Control Manual. Guide: Safety Management, NO.
CDC-22, Atlanta, GA. April 30, 1976 "Decontamination of Laboratory
Sink Drains to Remove Azide Salts."
6. Clinical and Laboratory Standards Institute (CLSI). Protection of
Laboratory Workers from Occupationally Acquired Infections; Approved
Guideline-Fourth Edition CLSI document M29-A4 Wayne, PA 2014.
*VP Echelon Series antibodies are developed solely by Biocare Medical
LLC and do not imply approval or endorsement of Biocare’s antibodies
by Ventana Medical Systems, Inc. Biocare and Ventana are not
affiliated, associated or related in any way. Ventana®, BenchMark®,
ultraView and OptiView are trademarks of Roche.
Rev: 062117
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