rev:   May 24, 2004

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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.

CD25 Antibodies

Mouse anti-human CD25 antibodies

-3 clones  

RDI-M1335clb              CLB-IL2R/1,TB-30       IV            mIgG2b     purified     $375.00

RDI-M1680clb                 " "                                "                "              FITC        $375.00

RDI-M1728clb                 " "                                "                "              PE             $438.00 RDI-CBL151BT               " "                                "               "               Biotin        $375.00

RDI-CBL151PC5            ""                                  "               "               R-PE/CY5  $469.00

RDI-cd25abm-B6           7G7B6                          IV           mIgG2a       purified, FITC, BT or PE

Catalog#: RDI-CD25abm-B6FT    $375.00/vial FITC label

Package Size: 120 tests FITC labeled in 0.2ml 50mM sodium phosphate pH 7.5, 500mM Potassium Chloride, 150mM NaCl, 5% glycerol, 0.2% BSA and 0.04% NaN3.

Species: mouse IgG2ak

Antigen: stimulated PBMC

Source: purified from tissue culture supernatant by Protein A, then conjugated to FITC (further purified by desalting column). Antibody conjugate is approx 0.25mg/ml F:P 13

LOT#

CLONE: 7G7B6

Activity: Antibody 7G7B6 recognizes epitope B of the IL-2 alpha receptor of about 55 kd (1,6). Antibody 7G7B6 does not block IL-2 binding to its receptor (1). Expression of human CD25 (the alpha subunit of the IL-2 receptor complex) is dramatically upregulated on activated T cells, B cells, NK cells and macrophages (3). Antibodies to CD25 are being studied for potential use as an immunodirected therapy to selectively attack (kill) CD25 expressing cells (4).

Application: direct flow cytometry

-titers must be determined for each application

Batch data: Dilution 1:50, use 80-ul/test as below: -cultured human CEM were stimulated overnight in the presence of PMA (10ng/ml) and ionomycin (1uM). 500K cells per tube were washed and incubated 45 minutes on ice with with 80ul of anti-CD25/FT at a dilution factor of 1:50. Cells were washed three times, fixed and analyzed on a BD FACS Calibur. cells stained positive with a mean shift of 1.5 log10 fluorescent units when compared to a Mouse IgG2aFT negative control (cat#RDI-MIGG2aFTX2 $188.00) at a similar concentration. continued on back side Binding was 83% blocked when cells were preincubated 10 minutes with 20ul of a 0.5mg/ml anti-CD25 antibody.

Storage: Store at 4 Deg C. DO NOT FREEZE. Protect from light.

Precautions: For In vitro research Use Only. Not for use in or on humans or animals or for diagnostics.

References:

1. L.A. Rubin, et al, (1985) Hybridoma 4:91-102.

2. B. Boutin, et al, (1989) Neuropediatrics 20: 202-206.

3. T.A. Waldmann, et al, (1989) Annu Rev Biochem 58: 875-911.

4. T.A. Waldmann, et al, (1995) Blood 86: 4063-4075.

5. U.S. Patent No. 4,707,443.

6. Leukocyte Typing IV (W. Knapp, et al, eds.) Oxford University Press, Oxford, (1989) p. 403-406.

It is the responsibility of the user to comply with all local/state and Federal rules in the use of this product. We are not responsible for any patent infringements that might result with the use of or derivation of this product.

Recommended FITC Isotype Control cat#RDI-MIGG1FTX2 $188.00/vial 100ug

Bulk Quotes on Request

mouse anti-human CD25   clone 7G7B6  , mIgG2a

cat#RDI-CD25abm-B6        $312.00/100ug purified

cat#RDI-CD25abm-B6PE    $469.00/120T PE conjugated

cat#RDI-CD25abm-B6FT     $375.00/120T FITC conjugated

cat#RDI-CD25abm-B6BT    $375.00/100ug Biotin conj

-unconjugated forms: suitable for flow cytometry, immunoprecipitation, acetone fixed frozen sections

PeliCluster  CD25

CAT#RDI- M1335clb

Test/vial 200

Clone CLB-IL2R, TB-30

This clone has been derived from hybridization of SP2/0 cells with spleen cells of a BALB/c mouse immunized with PHA stimulated T lymphocytes. This antibody has been clustered to in one of the international Workshop on Human White Cell differentiation Antigens

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 CD25- antigen (Interleukin-2 receptor).

The monoclonal antibody binds selectively to activated human lymphoblasts.

Resting human blood cells were found to be negative.

The monoclonal antibody is able to inhibit IL-2 driven T- cell proliferation.

Molecular mass 58 kDa.

Application Research on T- and B-cell activation.

Detection of cells infected with human T-cell leukaemia viruses I and II.

Studies of ligand receptor interactions.

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

References

1 Lier, R.A.W. van, et al., J. Immunology, 139, 2837 (1987).

2 Lier, R.A.W. van, et al., Eur. J. Immunology, 18, 167 (1988).

3 Lier, R.A.W. van, et al.; J. Immunology, 139, 1589 (1987).

4 Dongen, J.J.M. van, et al.; Neth. J. of Med., 33, 298 (1988).

5 Lier, R.A.W. van, et al., J. Immunology, 68, 45 (1987).

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

PeliCluster  CD25 F

CAT#RDI- M1680clb

Test/vial 100

Clone CLB-IL2R, TB-30

This clone has been derived from hybridization of SP2/0 cells with spleen cells of a BALB/c mouse immunized with PHA stimulated T lymphocytes. This antibody has been clustered to in one of the international Workshop on Human White Cell differentiation Antigens

Isotype Mouse, IgG2b.

Source Ascites fluid of tumour bearing BALB/c mice.

Purification Ammoniumsulphate precipitation and ion exchange chromatography.

Conjugation Conjugated with fluorescein iso thiocyanate isomer 1 (FITC).

Molecular F/P ratio between 5.5 - 9.5.

Packing Each vial contains 1 ml FITC conjugated monoclonal antibody and 10 mg BSA in PBS.

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 CD25- antigen (Interleukin-2 receptor).

The monoclonal antibody binds selectively to activated human lymphoblasts.

Resting human blood cells were found to be negative.

The monoclonal antibody is able to inhibit IL-2 driven T- cell proliferation.

Molecular mass 58 kDa.

Application Research on T- and B-cell activation.

Detection of cells infected with human T-cell leukaemia viruses I and II.

Studies of ligand receptor interactions.

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

References

1 Lier, R.A.W. van, et al., J. Immunology, 139, 2837 (1987).

2 Lier, R.A.W. van, et al., Eur. J. Immunology, 18, 167 (1988).

3 Lier, R.A.W. van, et al.; J. Immunology, 139, 1589 (1987).

4 Dongen, J.J.M. van, et al.; Neth. J. of Med., 33, 298 (1988).

5 Lier, R.A.W. van, et al., J. Immunology, 68, 45 (1987).

For research only! Not for human use! M1680\01 0708961448

FOR IN VITRO RESEARCH USE ONLY

PRODUCT CODE :RDI-CBL151  $375.00/vial

Also available FITC labeled cat#RDI-CBL151FT     $375.00/vial 100 tests

                          PE labeled cat#RDI-CBL151PE     $469.00/vial 100 tests

MOUSE MONOCLONAL

CLONE             Tu69

ISOTYPE          IgG1

IMMUNIZING ANTIGEN    Activated human T-cells

FUSION PARTNER    Sp2/0 myeloma cell line

SOURCE     Tissue culture supernatant

PURIFICATION METHOD    Protein A affinity chromatography

SPECIFICITY

This antibody reacts with the b chain of the interleukin-2 receptor (55 kDa glycoprotein), which is associated with human alloactivated T lymphocytes (FC2) kept in conditioned medium. The antigen is expressed on activated T and B lymphocytes and monocytes/macrophages.

Antigen:

Con A activated lymphocytes 67 + 27%

PHA activated lymphocytes 67 + 27%

Thymocytes 12+ 5%

B cells (E-) < 1%

T cells (E+) < 1%

Monocytes < 1%

Granulocytes < 1%

APPLICATIONS

* T cell leukaemia marker

* Early indications of acute rejection in renal transplantation

* Activity marker in some autoimmune diseases

* Biological analysis of the IL-2 receptor.

* Study of endocytosis and of the influence of IL-2 ligand on T cell physiology.

* Inhibition of the IL-2 dependent proliferation of T lymphocytes

*Identification of CD25 positive cells on acetone fixed frozen (NOT FOR FORMALIN FIXED PARAFFIN) paraffin wax embedded tissue sections

(using approx 20ug/ml, with pronase pretreatment) ABC detection, Lymphocytes scattered through the germinal centre, mantle and Tzones were labeled together with what looked like some monocytes. Pos control human tonsil. Antigen is susceptible to fixation and tissue processing.

* Identification of CD25 positive cells on acetone fixed cryostat sections (no pretreatemnt required, approx 1:100 dilution)

REFERENCES

Leucocyte Typing III, Oxford University Press (1987)

Leucocyte Typing IV, Oxford University Press (1990)

Waldemann, T. A. et al. Ann. Rev. Biochem. 58, 875-911 (1989

PRESENTATION The monoclonal is presented at a concentration of 200µg/2ml in phosphate buffered saline containing 10mM sodium azide and 1mg/ml bovine serum albumin. We recommend that each laboratory determine an optimum working titre for use in its particular application.

STORAGE For use within 1 month of purchase store at +4oC, for long term storage aliquot antibody into small volumes and store at -20oC.

Also available: Fitc labeled    cat#RDI-CBL151FT     $375.00/100T

                          PE lableled     cat#RDI-CBL151PE     $469.00/100T

                      Biotin labeled     cat#RDI-CBL151BT     $375.00/100T

                              R-PE/CY5   cat#RDI-CBL151PC5   $469.00/100T

Also available  in bulk milligram sizes-quotes 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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