rev:Nov 18, 1996

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CD CLUSTERED ANTIBODIES  

(anti-Human and others as indicated)

NOTE: 10% Discount on purchases of  3 or more any combination of CD antibodies. Please inquire for bulk or custom formulations (without preservative or carrier).

RDI Division of Fitzgerald Industries Intl  offers a wide line of  antibodies. Since no one antibody works best for all applications (flow cytometry, neutralization, blotting, histochemistry, ELISA, etc), we offer many different types of antibodies to help solve this problem. Please inquire for other applications or types of antibodies not listed below. All products are for in vitro research use only-not for use in or on humans or animals-not for use in diagnostics. Price/availability/specifications subject to change without notice.


CD3  Antibodies  (Paraffin Sections)


DATA SHEET CD3:Mouse Monoclonal Antibody (Paraffin Sections)


cat# RDI-CD3abm-PS1     $650.00/vial


Specificity: Human CD3E


Clone: PS1


Ig Class: IgG2a


Antigen: fusion protein to the epsilon chain of CD3


Hybridoma partner: Mouse myeloma (p3-NS1-Ag4-1).


Preparation: Lyophilised tissue culture supernatant containing 15mM sodium azide. Reconstitute with 1ml of sterile distilled water.


USE: Effective on paraffin wax embedded tissue using 0.01M sodium citrate unmasking solution ph 6.0 combined with the high temperature antigen unmasking technique


Protocol: Immunohistochemistry:Typical working dilution 1:100 - 1:200. High temperature antigen unmasking technique using 0.01M Na Citrate. 60 Minutes incubation at 25'C. Standard ABC technique.


Western blotting: Typical working dilution 1:100. Also suitable for ELISA


Positive controls: Tonsil


Staing pattern: Membrane


Storage and stability: Store unopened lyophilized antibody at 4'C. Under these conditions, there is no significant loss in product performance up to the expiry date indicated on the vial label. The reconstituted antibody is stable for at least two months when stored at 4'C. For long term storage, aliquot in non frost freze freezer, Avoid repeated freeze thaw cycles. Prepare fresh working dilutions daily.


Refs: Journal of Pathology. 173:303-307 (1994)


Stricly for in vitro research use only-Not for use in or on humans or animals-not for use in diagnostics


Sample High Temperature Unmasking Technqiue for Paraffin Sections:

     Buffer=1mM EDTA (ph 8.0) or 0.01M sodium Citrate Buffer (pH 6.0)


1) Cut and mount sections on slides coated with "apes"


2) Deparrafinize sections and rehydrate to distilled water


3) Bring 1mM EDTA* (pH 8.0) to a boil in a Prestige stainless steel pressure cooker, using a hot plate. Cover but do not lock lid. *or 0.01M sodium citrate buffer (pH 6.0)


4) Position slides into metal staining racks and lower into pressure cooker ensuring slides are well immersed in buffer Lock the lid. The small valve will rise.


5) When the pressure indicator valve (the large one) has risen after about 4 minutes, Incubate sections for 1 minute.


6) Remove pressure cooker from heat source and run under cold water with lid on. When the small valve sinks open lid and remove slides and place immediately into distilled water. DO NOT OPEN LID UNTIL THE SMALL VALVE SINKS.


7) Wash sections in TBS buffer (pH 7.6) for 1 X 5 minutes


8) Place sections in 1.5% hydrogen peroxide/methanol for 10 minutes.


9) Wash sections in distilled water for 2 X 5 minutes, then wash sections in TBS buffer for 2 X 5 minutes.


10) Place sections in normal serum for 20 minutes.


11) Cover sections with primary antibody. (Optimum dilution, incubation time and temperature must be determined for each antibody and in each lab).


12) Wash in TBS buffer for 2 X 5 minutes


13) Incubate sections in secondary antibody for 30 minutes


14) Wash in TBS buffer for 2 X 5 minutes


15) Incubate slides in ABC complex for 30 minutes


16) Wash in TBS buffer for 2 X 5 minutes


17) Incubate slides in DAB


18) Wash in distilled water for 2 X 5 minutes


19) Counterstain with haematoxylin (if required), dehydrate, coverslip, and mount



Material:

APES (3-aminopropyltriethoxysilane) cat#A3648 Sigma

To avoid sections becoming detached, sections should be mounted on "APES" covered slides, then dried at 37 DEG C overnight followed by drying at 56 DEG C for 60 minutes.


1mM EDTA (ph 8.0): Add 0.37g of EDTA (Sigma cat#E-5134) to 1 liter f distilled   water. Adjust pH to 8.0 using 1.0M sodium hydroxide.


0.01M Sodium Citrate Buffer (ph 6.0): Add 2.94g of tri-sodium citrate to 1 liter of distilled water. Adjust pH 6 using 1.0M HCL.


RDI Division of Fitzgerald Industries Intl

34 Junction Square Drive

Concord MA 01742-3049

USA

phone (978) 371-6446 or (800) 370-2222

fax     (978) 371-2266

EMAIL:antibodies@fitzgerald-fii.com

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