rev:January 3, 1997

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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.

CD42d Antibodies

Mouse anti-human CD42d antibodies

-1 clone 

CATALOG#                 CLONE#                  Wokshop    Host         Form        Price

RDI-M1637clb               CLB-SW16                   V            mIgG1       purified     $375.00

Production Specification: mouse monoclonal anti-HUMAN CD42d

PeliCluster  CD42d

cat#RDI- M1637clb   $375.00/vial

Test/vial 200

Clone CLB-SW16

This clone has been derived from hybridization of SP2/0 cells with spleen cells of a BALB/c mouse immunized with human platelets. This antibody has been clustered to CD42d in the fifth international Workshop on Human White Cell differentiation Antigens in Boston (1993).

Isotype Mouse IgG1.

Source Ascites fluid of tumour bearing BALB/c mice.

Purification Ammoniumsulphate precipitation and ion exchange chromatography.

Packing Each vial contains 1 ml with approximately 0.2 mg/ml monoclonal antibody and 10 mg BSA in 20 mM TRIS and 150 mM NaCl, pH 8.0.

Preservative Mertiolate (0.001%).

Storage and stability Monoclonal antibodies should be stored in the dark at 2-8°C. The reagent is stable until the expiry date stated on the vial label.

Major reactivity The monoclonal antibody is directed against the GP V which is expressed on human platelets. The monoclonal antibody reacts with human platelets. It has been shown that the glycoproteins GP V and GP Ib-GP IX form a noncovalent complex in the platelet membrane. The monoclonal antibody does not react with human lymphocytes, granulocytes, monocytes and erythrocytes.

Molecular mass 82 kDa.

Application Functional studies on cells.

The monoclonal antibody does not inhibit the von Willebrand factor-mediated agglutination of fixed platelets induced by 1.25 mg/ml of ristocetin in the presence of human plasma.

Methods Indirect immunofluorescence staining with analysis by flowcytometry or fluorescence microscopy.

References 1. Modderman, P.E. et al., J. of Biological Chemistry, 267, 364 (1992).

Application  FOR AN INDIRECT IMMUNOFLUORESCENCE TECHNIQUE

Reagents

- FITC-conjugated goat anti-mouse immunoglobulin antiserum (M1204).

  Remove aggregates by centrifugation at 1000 x g for 10 minutes.

- Buffer: Phosphate Buffered Saline (PBS), containing 0.2% Bovine Serum Albumin (BSA).

- PFA: Para-Formaldehyde 1%, pH 7.2, containing 0.2% BSA.

- Microtiter plates (96 wells, V bottom) or tubes.

Procedure

1   Fixate the cells with PFA 1% and wash them with buffer.

2   Prepare a mononuclear cell suspension with a concentration of 1 x 10 7 cells/ml in buffer.

3   Add 45 µl of cell suspension to the microtiter wells or tubes.

4   Add 5 µl monoclonal antibody to the microtiter wells or tubes and mix gently.

5   Incubate for 30 minutes at 2-8 C.

6   Add 100 µl buffer to the microtiter wells or 2 ml buffer to the tubes and centrifuge at 500 x g for 5 minutes.

7   Aspirate the supernatant from the cell pellet and resuspend the cells.

8   Add 200 µl buffer to the microtiter wells or 2 ml buffer to the tubes and centrifuge at 500 x g for 5 minutes.

9   Aspirate the supernatant from the cell pellet and resuspend the cells.

10 Add 50 µl FITC-conjugated goat anti-mouse immunoglobulin serum, solid phase adsorbed with human immunoglobulins,       diluted 1:80.

11  Incubate for 30 minutes at 2-8 C.

12  Repeat step 6-9.

13  Add 200 µl buffer.

14  The number of positive cells can be determined by flowcytometry analysis or by means of fluorescence microscopy.

NOTE:Care should be taken when drawing blood to avoid activation of platelets.

Platelets activation markers are weakly expressed on platelets prepared from directly fixed normal whole blood.

The expression is maximal on thrombin (1 U/ml) stimulated washed platelets.

FOR IN VITRO RESEARCH USE ONLY

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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