rev:   February 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 monoclonal 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.

CD10 Antibodies

Mouse anti-human CD10 antibodies

-4 clones     CLB-CALLA/1,4F9  (purified, FITC & PE)

                    Mem 78 (purified & PE)

                    RF-AL3 (purified & FITC)

                    56C6

CATALOG#                 CLONE#                Workshop  Host         Form        Price

RDI-M1337clb              CLB-CALLA/1,4F9       IV        mIgG2a    purified     $375.00

RDI-M1603clb                 " "                                "              "           FITC        $375.00

RDI-M1692clb                 " "                                "              "           PE            $438.00

RDI-CBL142                 Mem78                          IV         mIgG1     purified     $438.00

RDI-CBL142PE               " "                                "              "           PE            $531.00

RDI-CBL152FT                " "   **                         --             "           FITC        $375.00

             ** (Cytolytic complement fixing clone)

RDI-CD10abm-270         56C6                            --           mIgG1      TCS        $750.00/1ml (paraffin)

(not discountable)

PeliCluster CD10

cat#RDI- M1337clb  $375.00/vial

Test/vial 200

Clone CLB-CALLA/1, 4F9

This clone has been derived from hybridization of SP2/0 cells with spleen cells of a (BALB/c x A/J) mouse immunized with cells of a patient with Acute Lymphocytic Leukaemia of the c-ALL type. This antibody has been clustered to CD10 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 CD10- antigen (CALL-antigen), which is expressed on human lymphoblasts. The antibody reacts with early B lymphocytes (stem-cell, pre-B) and with the stem-cell of the lymphocyte lineage and immature thymocytes. Lymphoblasts of a patient with an Acute Lymphocytic Leukaemia of the c-ALL type were found to be positive. Normal B- and T lymphocytes, monocytes and platelets were found to be negative.

Molecular mass 100 kDa.

Application Characterization of non-T (common) Acute Lymphoblastic Leukaemias.

Analysis of early stages of haemopoietic differentiation.

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

References 1. Reinherz, E.L., Haynes, B.F., Nadler, L.M., Bernstein, I.D., Leukocyte Typing II, Springer Verlag, 2, New York (1985).

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

RDI-M1692clb                 " "                                "              "           PE            $438.00

ANTI-HUMAN CD10 CALLA ANTIGEN

PRODUCT CODE :RDI-CBL142       $438.00/vial 200ug
Also available PE labeled cat#RDI-CBL142PE    $531.00/vial 100 tests


CLONE: MEM 78

ISOTYPE: IgG1

SOURCE: Mouse ascitic fluid.

PURIFICATION METHOD: Protein A affinity chromatography.

SPECIFICITY: 95 kD antigen (CALLA) expressed on common acute lymphoblastic leukaemia cells which is seen on early B-lymphocytes and with stem cells of the lymphocyte lineage and immature thymocytes. The antigen is a neutral endopeptidase.

Antigen distribution: Granulocytes > 95%

                               cALL > 95%

                               Peripheral blood lymphocytes < 1%

                               T-cells (Ig+) < 1%

                               B-cells (Ig-) < 1%

                               Monocytes < 1%

                               Thymocytes < 1%

                               NK-cells (CD16+) < 1%

APPLICATIONS:

* The antibody can be used to define malignant cells designated as common acute lymphoblastic leukaemia cells

* The antibody works on fixed cells (not paraffin sections)

REFERENCES: Leucocyte Typing Workshop IV, Oxford University Press (1990).

                          Horejsi, V. et al. Fol. Biol. (Prague) 34, 23-24 (1988).

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

cat# RDI-CD10abm-270   $750.00/vial

Specificity: Human CD10 antigen

Clone: 56C6

Ig Class: IgG1

Antigen: prokaryotic recombinant protein corresponding to a portion of the extracellular domain of CD10 glycoprotein

Hybridoma partner: Mouse myeloma (p3-NS1-Ag4-1).

Preparation: Lyophilized tissue culture supernatant containing 15mM sodium azide. Reconstitute with 1ml of sterile distilled water.

USE: Effective on paraffin wax embedded tissue using 0.01M citrate buffer solution combined with the high temperature antigen unmasking technique

         frozen sections:acetone fixation

Protocol: Immunohistochemistry:Typical working dilution 1:40-1:80 after high temperature antigen unmasking technique using 0.01M Citrate pH6.0. 60 Minutes incubation at 25'C. Standard ABC technique.

Western blotting: 1:50-100 with chemiluminescence detection

Positive controls: small intestine or kidney

Staining pattern: Membrane

Storage and stability: Store unopened lyophilized antibody at 4'C. Under these conditions, there is no significant loss in product performance up to the expiry date indicated on the vial label. The reconstituted antibody is stable for at least two months when stored at 4'C. For long term storage, aliquot in non frost freze freezer, Avoid repeated freeze thaw cycles. Prepare fresh working dilutions daily.

ref: -American Journal of Pathology. 154(1):77-82 (1999)

Stricly for in vitro research use only-Not for use in or on humans or animals-not for use in diagnostics

CD10 is a 100kd cell surface metalloendopeptidase which inactivates a varitey of biologically active peptides. It was initially identified as the common acute lymphoblastic leukaemia antigen (CALLA) and considered to be tumour-specific. Sunsequent studies, however, have been shown that CD10 is expressed on the surface of a wide variety of normal and neoplastic cells. In other lymphoid malignancies, CD10 is expressed on cells of lymphobladtic, Burkitt's and follicular germinal centre lymphopmas, and on cells from patients with chronic myelocytic leukemia (CML) in lymphoid blast crisis. CD10 has been identified on the surface of normal early lymphoid progenitor cells, immature B cells within adult bone marrow and germinal centre B cells within lymphoid tissue. It is also expressed in various non-lymphoid cells and tissues, such as breast myoepithelial cells, bile canaliculi, fibroblasts, with especially high expression on the brush border and gut epithelial cells.

Sample High Temperature Unmasking Technqiue for Paraffin Sections:  0.01M sodium Citrate Buffer (pH 6.0)

1) Cut and mount sections on slides coated with "apes"

2) Deparrafinize sections and rehydrate to distilled water

3) Bring *or 0.01M sodium citrate buffer (pH 6.0) to a boil in a Prestige/Presto stainless steel pressure cooker, using a hot plate. Cover but do not lock lid.

4) Position slides into metal staining racks and lower into pressure cooker   ensuring slides are well immersed in buffer Lock the lid. The small valve   will rise.

5) When the pressure indicator valve (the large one) has risen after about 4-5   minutes, Incubate sections for 1 minute.

6) Remove pressure cooker from heat source and run under cold water with lid   on. When the small valve sinks open lid and remove slides and place   immediately into distilled water. DO NOT OPEN LID UNTIL THE SMALL VALVE   Sinks or Vent Cover Lock Drops (see manual with your particular unit).

7) Wash sections PBS buffer (10mM phosphate, 0.15M NaCl pH 7.5) for 1 X 5   minutes (or 100mM TRIS, 0.15M NaCl, pH 7.5)

8) Place sections in 1.5% hydrogen peroxide/methanol for 10 minutes (or use other appropriate peroxidase blocking procedure).

9) Wash sections in distilled water for 2 X 5 minutes, then wash sections in PBS buffer for 2 X 5 minutes.

10) Place sections in normal serum for 20 minutes.

11) Cover sections with primary antibody. (Optimum dilution, incubation time and temperature must be determined for each antibody and in each lab).

12) Wash in PBS buffer for 2 X 5 minutes

13) Incubate sections in secondary antibody for 30 minutes

14) Wash in PBS buffer for 2 X 5 minutes

15) Incubate slides in ABC complex for 30 minutes

16) Wash in PBS buffer for 2 X 5 minutes

17) Incubate slides in DAB or other suitable peroxidase substrate

18) Wash in distilled water for 2 X 5 minutes

19) Counterstain with haematoxylin (if required), dehydrate, coverslip, and mount

Material:

APES (3-aminopropyltriethoxysilane) cat#A3648 Sigma

To avoid sections becoming detached, sections should be mounted on "APES"  covered slides, then dried at 37 DEG C overnight followed by drying at 56 DEG  C for 60 minutes.

0.01M Sodium Citrate Buffer (ph 6.0): Add 2.94g of tri-sodium citrate to 1  liter of distilled water. Adjust pH 6 using 1.0M HCL.

Tris: 10mmM Tris, 0.15M NaCl, pH 7.5.

PBS: 10mM Phosphate, 0.15M MaCl pH 7.5

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

Ordering terms

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