rev:  September 9, 2003

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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.


CD13 Antibodies


Mouse anti-human CD13 antibodies

-3 clones     CLB-mon-gran/2,Q20 (purified , FITC or PE)

                    WM15 (purified, FITC & PE, & RPE-CY5)  22A5 (purified, FITC, PE or Biotin)


CATALOG#                 CLONE#                  Workshop    Host         Form        Price

RDI-M1387clb              CLB-mon-gran/2,Q20     III           mIgG2a     purified   $375.00

RDI-M1568clb               " "                                   " "             "             FITC      $375.00

RDI-M1726clb               " ":                                   " "            "              PE          $438.00 

RDI-CBL169PC5            ""                                     "              "             RPE-CY5 $500.00

RDI-CD13abm-A5         22A5                               --            mIgG2a     purified    $375.00

RDI-CD13abm-A5FT        "                                    --              "              FITC        $375.00

RDI-CD13abm-A5PE        "                                   --              "              PE           $500.00

RDI-CD13abm-A5BT       "                                    --              "              Biotin      $375.00


Product Specification: mouse monoclonal anti-human CD13


PeliCluster  CD13


cat#RDI- M1387clb    $375.00/vial


Test/vial 200

Clone CLB-mon-gran/2, Q20

This clone has been derived from hybridization of SP2/0 cells with spleen cells of a BALB/c mouse immunized with human myeloblasts of a chronic myeloïd leukaemia patient in blast crisis. This antibody has been clustered to CD13 in the third International Workshop on Human White Cell differentiation Antigens in Oxford (1986).


Isotype Mouse, IgG2a.

Source Ascites fluid of tumor bearing BALB/c mice.

Purification Ammoniumsulphate precipitation and ion exchange chromatography.

Packing Each vial contains 1 ml with approximately 0.1 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 CD13-antigen, which is expressed on human monocytes, granulocytes and their precursors. The monoclonal antibody reacts with monocytes, granulocytes and with a large number of acute myeloïd leukaemias (the positivity of this marker relates to worse clinic prognosis)


Molecular mass 150 kDa.

Application Enumeration of granulocytes, monocytes and their precursors in peripheral blood.

Identification of granulocytes, monocytes and their precursors in lymphoid tissue.

Characterization of subtypes of leukaemias and lymphomas.

Research of calcium influx with monocytes.



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

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.

FOR IN VITRO RESEARCH USE ONLY


Product Specification: mouse monoclonal CD13 conjugated to Fluorescein


PeliCluster CD13 F



cat#RDI- M1568clb

Test/vial 100

Clone CLB-mon-gran/2, Q20


This clone has been derived from hybridization of SP2/0 cells with spleen cells of a BALB/c mouse immunized with human myeloblasts of a chronic myeloïd leukaemia patient in blast crisis. This antibody has been clustered to CD13 in the third International Workshop on Human White Cell differentiation Antigens in Oxford (1986).

Isotype Mouse, IgG2a.

Source Ascites fluid of tumor bearing BALB/c mice.

Purification Ammoniumsulphate precipitation and ion exchange chromatography.


Conjugation Conjugated with fluorescein iso tiocyanate 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 CD13-antigen, which is expressed on human monocytes, granulocytes and their precursors. The monoclonal antibody reacts with monocytes, granulocytes and with a large number of acute myeloïd leukaemias (the positivity of this marker relates to worse clinic prognosis)


Molecular mass 150 kDa.


Application Enumeration of granulocytes, monocytes and their precursors in peripheral blood.

Characterization of subtypes of leukaemias and lymphomas.

Research of calcium influx with monocytes.


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


Application in direct Immunofluorescence techniques

Method with ficoll purified cells


Reagents and materials

- FITC-conjugated monoclonal antibodies.

- Buffer: Phosphate Buffered Saline (PBS), containing 0.2% Bovine Serum Albumine (BSA)

- PFA: Para-Formaldehyde 1%, pH 7.2,  containing 0.2% BSA

- Microtiter plates (96 wells, V bottom) or tybes.


Procedure

1 Prepare a mononuclear cell suspension with a concentratation of 1 x 10 7 cells/ml.

2 Ad 40 µl of cell suspension to microtiter wells or tubes.

3 Add 10 µl of the antibody to the microtiter wells or tubes and mix gently.

4 Incubate for 30 minutes at 2-8°C.

5 Add 150 µl buffer to the microtiter wells or 2 ml buffer to the tubes and centrifuge at 500 x g for 5 minutes.

6 Aspirate the supernatant from the cell pellet and resuspend the cells.

7 Add 200 µl buffer to the microtiter wells or 2 ml buffer to the tubes and centrifuge at 500 x g for 5 minutes.

8 Aspirate the supernatant from the cell pellet and resuspend the cells.

9 Cytoflow fluorometer analysis:

    Add 200 µl buffer to the microtiter wells and transfer this final cell suspension to appropriate testtubes, or add 200 µl             buffer to the tubes.

10 If analysis within 8 hours is not possible add at no. 9, instead of buffer, 200 µl PFA 1%.



Fig. 1

Fluorescense profile of ficoll purified normal cells

(Scatter gates set on the granulocyte fraction)


WHOLE blood method

Due to sophisticated laser flowcytometry we are now able to distinguish populations of human lymphocytes, monocytes and granulocytes by their characteristics in different light scattering. By gating on the appropriate cell cluster (Fig.2) we can easily determine mononuclear cells, which have been immunofluorescently stained with monoclonal antibodies, and enumerate them.

Reagents and materials

- FITC-conjugated monoclonal antibodies.

- Lysing solution (NH 4Cl, pH 7.2).

- Buffer: PBS, containing 0.2 % BSA.

- PFA: Para-Formaldehyde 1%, pH 7.2, containing 0.2 % BSA.

- Tubes.


Procedure

1 Draw blood into a blood collection tube containing EDTA.

2 Deliver 100 µl (*) of well mixed whole blood to the bottom of the testtube.

3 Add 10 µl of the undiluted antibody to the bottom of the testtube, and mix gently.

4 Incubate for 30 minutes at 2-8°C or in an ice-water bath.

5 Mix the tubes and add 2 ml of lysing solution.

6 Incubate for 3-5 minutes at room temperature.

7 When lysing is completed, centrifuge the tubes at 500 x g for 5 minutes.

8 Aspirate the supernatant from the cell pellet, resuspend the cells in 1 ml buffer.

Place the tubes in the ice-water bath.

9 Analyse the samples within 8 hours.

If analysis within 8 hours is not possible centrifuge the tubes at 500 x g. Aspirate the supernatant from the cell pellet and resuspend the cells in 1 ml PFA 1%.

Fig. 2:

Scatter patern of a whole normal blood sample

R2 - lymphocytes R3 - monocytes R4 - granulocytes

Note:Use our special negative control to determine background fluorescence produced due to Fc binding capacities by mononuclear cells.The concentration and F/P ratio of our control has been adjusted to our FITC-conjugated monoclonal antibodies.

FOR IN VITRO RESEARCH USE ONLY


Mouse ANTI-human CD13 ANTIGEN


PRODUCT CODE :RDI-CBL169     $438.00/vial

  Also available: FITC labeled cat#RDI-CBL169FT    $395.00/vial 100 Tests

                             PE labeled cat#RDI-CBL169PE    $500.00/vial\

                             RPE-CY5 cat#RDI-CBL169PC5   $500.00/100T

CLONE    WM 15

ISOTYPE    IgG1

SOURCE    Mouse ascitic fluid

PURIFICATION METHOD   Ion exchange chromatography

SPECIFICITY

The CD13 antigen is present on many non-haemopoietic tissues and represents an integral single chain membrane glycoprotein of 150 kDa. Present on monocytes and weakly expressed on granulocytes, it is only present on a small proportion of normal bone marrow cells. These comprise a major portion of the normal granulocyte-macrophage progenitor population. CD13 is a type III glycoprotein, the extracellular region of which contains the catalytic unit of aminopeptidase-N which is a zinc binding metalloprotease.


APPLICATIONS

*Phenotyping of leukaemias

*Immunoprecipitation studies of the CD13 moiety

*This antibody may be used to deplete plasma of its aminopeptidase-N activity since the plasma enzyme may be the soluble form of membrane derived CD13

*Studies of serum alanine aminopeptidase in hepatobiliary disease since the enzyme is an isoform of the CD13 antigen

KNOWN SPECIES CROSS REACTIVITY       recognizes rat

REFERENCES

Leucocyte Typing III, Oxford University Press (1987)

Leucocyte Typing IV, Oxford University Press (1989)

Bradstock, K. F. et al. Br. J. Haem. 61, 11-20 (1985)

Favaloro, E. J. et al. Clin. Chim. Acta 220, 81-90 (1993)

Favaloro, E. J. et al. Exp. Haematol. 21, 1695-1701 (1993)

LOT SPECIFIC DATA

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-CBL169FT $395.00/100T

PE labeled cat#RDI-CBL169PE $500.00/100T


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


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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