rev: November 8, 1999

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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.

CD49f Antibodies

Mouse anti-human CD49f  antibodies

-2 clones 

CATALOG#                 CLONE#                  Workshop    Host         Form        Price

RDI-M1566clb              CLB-GOH3                  IV            ratIgG2a    purified     $375.00

RDI-M1672clb                 "                                  "                 "              FITC       $375.00

RDI-CBL458                4F10                              --            mIgG2b      purified     $375.00

RDI-CBL458FT              " "                                --              "               FITC       $375.00

RDI-CBL458PE             " "                                 --              "               PE           $469.00

Product Specification: rat monoclonal  anti-human CD49f.

PeliCluster  CD49f

cat#RDI- M1566clb

Test/vial 200

Clone NKI-GoH3

This clone has been derived from hybridization of SP2/0 cells with spleen cells of Sprague Dawley rats immunized with cells from mammary tumours induced in a BALB/c mouse by the C3H mouse mammary tumour virus. This antibody has been clustered to CD49f in the fifth international Workshop on Human White Cell differentiation Antigens in Boston (1993).

Isotype Rat IgG2a.

Source Culture supernatant..

Purification 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 CD49f- antigen (GP Ic or VLA-6 alpha chain), which can form distinct complexes with either the CD29-antigen (GP IIa or VLA beta chain), resulting in the VLA-6 (alpha-6 beta- 1) complex, which is expressed on human platelets, or with the beta-4 integrin resulting in the alpha-6 beta-4 complex, which is expressed on various human epithelial cells. The monoclonal antibody reacts with platelets, megakaryocytes, T lymphocytes and common acute lymphoblastic leukaemia cells (alpha-6 beta-1). In immunohistology the monoclonal antibody reacts with epithelial cells of a variety of tissues, peripheral nerves, microvascular endothelial cells, placenta cyto- and syncytotrophoblasts.

Molecular mass 30/31, 120 kDa.

Application Functional studies on cells.

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

References

1. Sonnenberg, A. et al., J.Biol.Chem., 262, 10376 (1987).

2. Hemler, M.E. et al., J.Biol.Chem., 263, 7660 (1988).

3. Sonnenberg, A. et al., J.Biol.Chem., 263, 14030 (1988).

4. Hemler, M.E. et al., J.Biol.Chem., 264, 6529 (1989).

5. Sonnenberg, A. et al., Nature, 336, 487 (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.

- microplate 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 microplate wells or tubes.

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

5  Incubate for 30 minutes at 2-8 C.

6  Add 100 µl buffer to the microplate 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 microplate 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 RESEARCH USE ONLY

Mouse ANTI-HUMAN CDw49F ANTIGEN (Laminin receptor)

PRODUCT CODE : RDI- CBL 458     $375.00/vial 200ug

-also available FITC labeled cat#RDI-CBL458FT   $375.00/100 test vial

                          PE labeled cat#RDI-CBL458PE   $469.00/100 test vial

CLONE: 4F10

ISOTYPE: IgG2b

SOURCE: Tissue culture supernatent.

PURIFICATION METHOD: Protein A affinity chromatography.

SPECIFICITY: 4F10 is specific to the integrin a-6 chain which is expressed on epithelial and endothelial basement membrane. This VLA a-6 chain, which consists of a single disulphide linked 120 kDa subunit and a small doublet subunit of 30/31 kDa, is non covalently associated with the VLA ß-chain in VLA-6 or a6/ß1 complex or with integrin ß4 chain(a6/ß4).

APPLICATIONS:

* Immunohistological studies on frozen sections.

* Immunoprecipitation of the laminin receptor.

PRESENTATION: Each monoclonal is delivered in a sealed vial at a concentration of not less than 200µg/2ml in phosphate buffered saline containing 10mM sodium azide and 1mg/ml of bovine serum albumin. 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

also available in bulk with or without BSA and preservative.

-$1062.00/1 milligram in bulk. Larger quantities quoted on request. order cat#RDI-CBL459-1XP 1 milligram, NO BSA/NO AZIDE.

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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