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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.

CD49e Antibodies

Mouse anti-human CD49e  antibodies

-1 clone  

CATALOG#                 CLONE#                  Workshop    Host         Form        Price

RDI-M1586clb              SAM-1                           V            mIgG2a     purified     $375.00

RDI-CBL497FT              " "                                  "                "           FITC         $438.00

RDI-CBL497PE              " "                                  "                "           PE             $500.00

Product Specification: mouse monoclonal anti-Human CD49e

PeliCluster  CD49e

cat#RDI- M1586clb

Test/vial 200

Clone NKI-SAM-1

This clone has been derived from hybridization of SP2/0 cells with spleen cells of a BALB/c mouse immunized with U937. This antibody has been clustered to CD49e in the fifth international Workshop on Human White Cell differentiation Antigens in Boston (1993).

Isotype Mouse IgG2b.

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 CD49e- antigen (GP Ic' or VLA-5 alpha chain), which can form distinct complexes with either the CD29-antigen (GP IIa or VLA beta chain), resulting in the VLA-5 (alpha-5 beta- 1) complex which is expressed on human platelets.

The monoclonal antibody reacts with platelets and monocytes.

The monoclonal antibody reacts weakly with T-cells and granulocytes.

The monoclonal antibody also reacts with various cell types of non-haemopoietic origin.

Molecular mass 17, 130 kDa.

Application Functional studies on cells.

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

References 1. Velde, A. te, et al., J. of Immunology, 140, 1548 (1988).

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. M1586

FOR IN VITRO RESEARCH USE ONLY

MOUSE ANTI-Human CDw49E monoclonal

PRODUCT CODE :RDI-CBL497                 $375.00/vial 200ug

available FITC labled cat#RDI-CBL497FT     $438.00/vial 100 tests

                  PE labeled cat#RDI-CBL497PE   $500.00/vial 100 tests

CLONE: Sam-1

ISOTYPE: IgG2b

Immunogen: U937 cells

Fusion Partner: SP2/0

SOURCE: Mouse ascitic fluid

PURIFICATION METHOD: Ammonium sulphate precipitation followed by ion exchange chromatography.

SPECIFICITY: The monoclonal antibody is directed against the CD49E-antigen (GP Ic or VLA-5 alpha chain) which can form distinct complexes with either the CD29antigen (GP IIa or VLA beta chain) resulting in the VLA-5 (alpha-5-beta-1) complex which is expressed on human platelets. The monoclonal antibody reacts with platelets and monocytes and also weakly with T cells and granulocytes. It also reacts with various cell types of non-haemopoietic oorigin.

APPLICATIONS:

Functional studies on cells of the platelet VLA-5 which functions as a fibronectin receptor and platelet adhesion to fibronectin is inhibited by Sam-1.

Indirect immunofluorescence staining with analysis by FACS or fluorescence microscopy.

REFERENCES: A. te Velde et al, J. of Immunology 140, 1548, 1988.

PRESENTATION: Each monoclonal is delivered in a sealed vial at a concentration of 200 ug/2ml with 10mm NaN3 and 1mg/ml BSA. We recommend that each laboratory determine an optimum working titre for use in its particular application.

STORAGE: Purified monoclonal antibodies are generally stable for long periods at 4oC. However repeated warming to room temperature and re-cooling may result in a loss of activity and hence effective working titre. It is recommended that monoclonal antibodies are aliquotted upon receipt and then stored at -20C. After thawing they should then be used as a stock solution for up to 60 days. Dilute working solutions are best prepared on the day of use.

For research use only. Not supplied for use in human diagnostic or therapeutic procedures

BULK material, no BSA, no azide , cat#RDI-CBL497-1XP $1125.00/1 milligram

Larger sizes and custom material quoted on request.

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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