rev: January 23, 2003
HOME (index HOME page)
Return (master CD antibody index page)
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.
CD8
Antibodies (Paraffin Sections)
cat# RDI-CD8abm-4B11 $688.00/vial 1ml
alos available as combination pack with
CD4 (paraffin clone)
cat#RDI-CD4CD8-PK $688.00/set 0.5ml each
Specificity: Human CD8
Clone: 4B11
Ig Class: IgG2b
Antigen: peptide corresponding to the alpha chain cytoplasmic portion of
the human CD8 molecule.
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 1mM EDTA pH 8.0 unmasking
solution combined with the high temperature antigen unmasking technique
- frozen sections:acetone fixed
Protocol: Immunohistochemistry:Typical working dilution 1:20 - 1:40. High temperature antigen unmasking technique using 1mM EDTA (pH 8.0). 60 Minutes incubation at 25'C. Standard ABC technique.
-Western blotting: Typical
working dilution 1:25. 60 minutes incubation at 25'C.
Positive controls: Tonsil
Stainig 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 free freezer, Avoid repeated freeze thaw cycles. Prepare
fresh working dilutions daily.
Refs: Journal of Clinical Pathology 45:1084-1088
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) Place sections in 0.5% hydrogen peroxide/methanol for 10 minutes.(or other
appropriate endogeneous peroxidase procedure). Wash sectionsin tap water.
4) Bring 1600ml 1mM EDTA* (pH 8.0) to a boil in a Prestige/Presto stainless steel pressure cooker, using a hot plate. Cover but do not lock lid. Follow operating instructions with your individual unit/microwave etc
5) Bring 1600ml of 1mM EDTA* (pH 8.0) to a boil in a Presto stainless steel pressure cooker,using a hot plate. Cover but do not lock lid.
6) Position slides into metal staining racks (do not place slides close together, uneven staining may occur) and lower into pressure cooker ensuring slides are well immersed in buffer Lock the lid. The air vent/cover and overpressure plug will rise.
7) When the pressure indicator valve (the large one) has risen after about 5 minutes, Incubate sections for 1 minute.
8) 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 Vent Cover Lock Drops OR PRESSURE HAS DROPPED.
9) Wash sections in PBS* buffer (pH 7.5) for 1 X 5 minutes
10) Wash sections in distilled water for 2 X 5 minutes, then wash sections in PBS buffer for 2 X 5 minutes.
11) Place sections in diluted normal serum for 10 minutes.
12) Cover sections with primary antibody. (Optimum dilution, incubation time and temperature must be determined for each antibody and in each lab).
13) Wash in PBS buffer for 2 X 5 minutes
14) Incubate sections in secondary antibody for 30 minutes
15) Wash in PBS buffer for 2 X 5 minutes
16) Incubate slides in ABC complex for 30 minutes
17) Wash in PBS buffer for 2 X 5 minutes
18) Incubate slides in DAB
19) Wash in distilled water for 2 X 5 minutes
20) 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 of distilled water. Adjust pH to 8.0 using 1.0M sodium hydroxide.
Tris: 100mmM Tris, 0.15M NaCl, pH 7.5.
PBS: 10mM Phosphate, 0.15M MaCl pH 7.5
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
RETURN (CD Antibody INDEX page)