rev: February 1, 2002

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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.

CD27 & CD27L Antibodies (monoclonals and polyclonals)

see also our compact Elisa kit to measure sCD27

PeliCluster CD27

CAT#RDI- M1455clb  $375.00

-also availabel FITC conjugated:   cat#RDI-M1764clb  $375.00/100T

Test/vial 200

Clone CLB-CD27/1, 9F4
This clone has been derived from hybridization of SP2/0 cells with spleen cells of a (BALB/c x A/J) mouse immunized with E-rosette positive human lymphocytes. This antibody has been clustered to CD27 in one of the international Workshop on Human White Cell differentiation Antigens.

Isotype Mouse IgG2a.

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 CD27- antigen (TP 55-antigen), which is expressed on a large subpopulation (75%) of peripheral blood T lymphocytes and most medullary thymocytes. Little binding was observed to normal human B-cells, CD3- negative leukaemic cells, B-cell derived and myeloid leukaemia.

Molecular mass 32, 55 kDa.

Application Identification of two functionally distinct subpopulations within the CD4+ T-cell subset..

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

References

1. Lier, R.A.W. van, et al., J.Immunol., 139, 1589 (1987).

2. Lier, R.A.W. van, et al., Eur.J.Immunol., 18, 811 (1988).

3. Borst, J., et al., Eur.J.Immunol., 19, 357 (1989).

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-Not for use in diagnostics

Mouse ANTI-CD27 ANTIGEN

PRODUCT CODE :RDI-CBL455   $375.00/vial 200ug - DISCONTINUED

CLONE: LT- 27

ISOTYPE: IgG2a

SOURCE: Mouse ascitic fluid.

PURIFICATION METHOD: Ion exchange chromatography following ammonium sulphate precipitation.

SPECIFICITY: The antibody precipitates a molecule of MW55kDa (120kDa in non-reduced conditions) designated the CD27 antigen. This antigen is expressed on activated T lymphocytes and belongs to a family of proteins associated with the nerve growth factor receptor. It is possible that the CD27 antigen is an analogue of the CD40 protein on activated B-cells..

APPLICATIONS: * Indirect immunofluorescence and FACS analysis for thymocytes and                                           peripheral T cells.

                               * EBV-transformed B-cell lines and a major T-cell subset.

REFERENCES Leucocyte Typing IV, Oxford University Press (1990)

PRESENTATION: Each monoclonal is delivered in a sealed vial at a concentration of 200µg/2ml in phosphate buffered saline solution containing 10mM sodium azide 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 -200C. 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 (1mg or more) available on request with or without sodium azide. $1125.00/1 mg. order cat#RDI-CBL455-1XP NO BSA/NO AZIDE

also available FITC labeled cat#RDI-CBL455FT $438.00/vial 100T

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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