rev:  October 3, 2001

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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.


CD36 Antibodies


Mouse anti-human CD36 antibodies

-3 clones  


CATALOG#                 CLONE#                  Workshop    Host         Form        Price

RDI-M1537clb               CLB-1VC7                    --           mIgG1      purified     $375.00

RDI-M1613clb                 " "                                 --             "             FITC        $375.00

RDI-CBL168                  SM0                               IV         mIgM        purified     $375.00

RDI-CBL168FT               " "                                  "             "             FITC        $375.00

RDI-CD36NL07              NL07 (CB38)               IV          mIgM       purified     $281.00


Product Specification: mouse monoclonal anti-human CD36

PeliCluster CD36

CAT#RDI- M1537clb

Test/vial 200

Clone CLB-IVC7

This clone has been derived from hybridization of SP2/0 cells with spleen cells of a BALB/c mouse immunized with human monocytes. This antibody has been clustered to CD36 in one of the international Workshop on Human White Cell differentiation Antigens.


Isotype Mouse IgG1.


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 CD36- antigen (gpIV also known as gpIIIb), which is expressed on human thrombocytes. The monoclonal antibody reacts with thrombocytes, monocytes, macrophages, erythroblasts and weak with B- cells. In immunohistology the monoclonal antibody reacts with some endothelial cells, adipocytes and the granular layer of the skin.


Molecular mass 90 kDa.

Application Functional studies on cells.

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

References

1. Asch, A.S. et al., J. Clin. Invest., 79, 1054 (1987).

2. Ockenhouse, C.F. et al., Science, 243, 1469 (1989).

3. Tandon, N.N. et al., J. Biol. Chem., 264, 7576 (1989).

4. Tandon, N.N. et al., J. Biol. Chem., 264, 7570 (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. M1537


FOR IN VITRO RESEARCH USE ONLY


Product Specification: mouse monoclonal CD36,conjugated to Fluorescein

PeliCluster  CD36 F

CAT#RDI- M1613clb

Test/vial 100

Clone CLB-IVC7

This clone has been derived from hybridization of SP2/0 cells with spleen cells of a BALB/c mouse immunized with human monocytes. This antibody has been clustered to CD36 in one of the international Workshop on Human White Cell differentiation Antigens.

Isotype Mouse IgG1.

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 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 CD36- antigen (gpIV also known as gpIIIb), which is expressed on human thrombocytes. The monoclonal antibody reacts with thrombocytes, monocytes, macrophages, erythroblasts and weak with B- cells. In immunohistology the monoclonal antibody reacts with some endothelial cells, adipocytes and the granular layer of the skin.


Molecular mass 90 kDa.

Application Functional studies on cells.

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


References

1. Asch, A.S. et al., J. Clin. Invest., 79, 1054 (1987).

2. Ockenhouse, C.F. et al., Science, 243, 1469 (1989).

3. Tandon, N.N. et al., J. Biol. Chem., 264, 7576 (1989).

4. Tandon, N.N. et al., J. Biol. Chem., 264, 7570 (1989).


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 monocyte 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. * This method was developed for blood samples with a normal white count. It may be necessary to adjust the quantity of blood for samples with very high or low white counts M1613/02 070896141

FOR IN VITRO RESEARCH USE ONLY


MOUSE ANTI-HUMAN CD36 ANTIGEN

PRODUCT CODE :RDI-CBL168     $375.00/vial 200ug purified
Also available FITC labeled cat#RDI-CBL168FT $375.00/100 Tests


CLONE: SMO

ISOTYPE: IgM

SOURCE: Mouse ascitic fluid

PURIFICATION METHOD: Ammonium sulphate precipitation followed by gel filtration.

SPECIFICITY: 90kDa molecular weight single chain membrane glycoprotein (GPIIIb) that is found on monocytes/macrophages and platelets.


APPLICATIONS:

* Studies of platelet and monocyte receptor for thrombospondin

* Studies of platelet collagen adhesion receptor

REFERENCES: Hogg, N. Immunol. 53, 753 (1985)

                           Leucocyte Typing IV, Oxford University Press (1989)

                          Greenwalt,D.E. et al. Blood 80 (5), 1105 - 15 (1992)


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 4 DEG C. After thawing they should then be used as a stock solution for up to 30 days. Dilute working solutions are best prepared on the day of use. DO NOT FREEZE IgM Isotype Antibody.

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

Available in bulk, with or without BSA and azide$1062.00/milligram order cat#RDI-CBL168-1XP Larger sizes quoted upon request


DATA SHEET:    mouse anti-human CD36 clone CB38/NL07

Catalog#: RDI-CD36NL07     $281.00/vial     $250.00/vial 3 or more


Package Size: 100ug purified in 0.2ml (0.5mg/ml) 50mM Tris-Hcl (pH 8.0), 0.15M NaCl and 0.1% sodium azide. (sodium azide is toxic, dilute azide compounds with copious amounts of water when disposing in drain to avoid accumulation of explosive deposits-avoid contact with acids.)

Prep: purified from tissue culture supernatant via Protein G affinity chromatography.

Species: mouse IgM

CLONE: CB38 also known as NL07

Workshop Code: Workshop IV, P106

Activity: Reacts with the 88kD glycoprotein IV (GPIV) the receptor for extracellular matrix proteins such as collagen and thrombospondin. CD36 is known to mediate the adhesion of Plasmodium falciparum. CD36 antigen is expressed on monocytes, platelets, endothelial cells, and some human tumor cell lines but not on lymphocytes and granulocytes. It is a very early marker of erythroid differentiation. CD36 antibody induces degranulation, release of ATP and   serotonin, increase in CA2+, and tyrosine phosphorylation of a substrate protein of 130kD.


Application: -Indirect immunofluorescence cell surface staining , Titer for each application


References: 1) Leucocyte Typing IV

                  2) Blood 82:3637

                  3) Current Studies in Hematology and Blood Transfusion 58:182

Storage: Store at 4 DEG C.

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



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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