Estrogen Receptor (ER) [SP1]

Estrogen Receptor (ER) [SP1]
Estrogen Receptor (ER) [SP1]
Concentrated and Prediluted Rabbit Monoclonal Antibody
Control Number: 902-301-040215
Catalog Number:
CRM 301 A, B, C
PRM 301 AA
OAR 301 T60
Description:
0.1, 0.5, 1.0 ml, concentrated
6.0, ml prediluted
60 tests, prediluted
Dilution:
1:50-1:100
Ready-to-use
Ready-to-use
Diluent:
Renoir Red
N/A
N/A
Intended Use:
For Research Use Only. Not for use in diagnostic procedures.
Summary and Explanation:
Human estrogen receptor (ER) is a 66 kDa protein that acts as an estrogen-dependent,
nuclear hormone receptor. Studies have shown ER is present in the nuclei of epithelial
cells in normal breast and endometrial tissues, as well as a subset of breast carcinomas.
The SP1 clone is a high affinity rabbit monoclonal antibody directed against an epitope
of the C-terminus of the ER protein. (1) SP1 has been shown to stain formalin-fixed
paraffin-embedded tissues. The robustness of SP1 has been demonstrated by successful
immunohistochemistry using lower temperatures for antigen retrieval (e.g. 80 or 95ºC),
and in some instances, staining can be obtained even without antigen retrieval. (2-3)
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. After labeling the
antigen with a primary antibody, an enzyme labeled polymer is then added to bind to
the primary antibody. This detection of the bound antibody is evidenced by a
colorimetric reaction.
Source: Rabbit monoclonal
Species Reactivity: Human; others not tested
Clone: SP1
Isotype: IgG
Total Protein Concentration: ~10 mg/ml. Call for lot specific Ig concentration.
Epitope/Antigen: ER
Cellular Localization: Nuclear
Positive Control: Breast carcinoma
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.
Staining Protocol Recommendations (manual use):
Peroxide Block: Block for 5 minutes with Biocare's Peroxidazed 1.
Pretreatment Solution (recommended): Diva
Pretreatment Protocol:
Heat Retrieval Method:
Preheat the retrieval solution to 95°C for 30 minutes in Biocare's Decloaking Chamber.
Then, place slides into the preheated solution and retrieve under pressure at 95°C for
40 minutes; alternatively, steam tissue sections for 45-60 minutes or use a water bath at
95°C for 40 minutes. Allow solution to cool for 20 minutes then wash in distilled
water.
Protein Block (Optional): Incubate for 5-10 minutes at RT with Biocare's Background
Punisher.
Primary Antibody: Incubate for 30 minutes at RT.
Probe: N/A
Polymer: Incubate for 30 minutes at RT with a secondary-conjugated polymer.
Chromogen:
Incubate for 5 minutes at RT with Biocare's DAB - OR - Incubate 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.
Technical Note:
This antibody has been standardized with Biocare's MACH 2 detection system. It can
also be used on an automated staining system and with other Biocare polymer detection
kits. Use TBS buffer for washing steps.
Protocol Recommendations (ONCORE Automated Slide Staining System):
OAR301 is intended for use with the ONCORE Automated Slide Staining System.
Refer to the ONCORE Automated Slide Staining System User Manual for specific
instructions on its use. Protocol parameters in the ONCORE Automated Slide Stainer
Protocol Editor should be programmed as follows:
Protocol Name: ER Rb
Protocol Template (Description): Rb HRP Template 1
Dewaxing (DS Option): DS2
Antigen Retrieval (AR Option): AR2, low pH; 95°C
Reagent Name, Time, Temp.: ER Rb, 30 min., 25°C
Limitations:
This product is provided for Research Use Only (RUO) and is not for use in diagnostic
procedures. Suitability for specific applications may vary and it is the responsibility of
the end user to determine the appropriate application for its use.
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 in 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/.
Technical Support:
Contact Biocare's Technical Support at 1-800-542-2002 for questions regarding this
product.
References:
1. Cheang MC, et al. Immunohistochemical detection using the new rabbit monoclonal
antibody SP1 of estrogen receptor in breast cancer is superior to mouse monoclonal
antibody 1D5 in predicting survival. J Clin Oncol. 2006 Dec;24(36):5637-44.
2. Rossi S, et al. Rabbit monoclonal antibodies: a comparative study between a novel
category of immunoreagents and the corresponding mouse monoclonal antibodies. Am
J Clin Pathol. 2005 Aug;124(2):295-302.
3. Cano G, et al. Estimation of hormone receptor status in fine-needle aspirates and
paraffin-embedded sections from breast cancer using the novel rabbit monoclonal
antibodies SP1 and SP2. Diagn Cytopathol. 2003 Oct; 29(4):207-11.
Page 1 of 2
Estrogen Receptor (ER) [SP1]
Concentrated and Prediluted Rabbit Monoclonal Antibody
Control Number: 902-301-040215
References Cont'd:
4. Rocha R, et al. Rabbit monoclonal antibodies show higher sensitivity than mouse
monoclonals for estrogen and progesterone receptor evaluation in breast cancer by
immunohistochemstry. Pathol Res Pract. 2008; 204(9):655-62.
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.
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