rev:  January 8, 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.

CD38 Antibodies

CATALOG#                 CLONE#                  Workshop    Host         Form        Price

RDI-M1642clb               CLB-1D5                       --           mIgM        purified     $375.00

RDI-CD38HIT2              HIT2                               III            mIgG1k   purified    $315.00

RDI-CD38abm-290      SPC32                             ---           mIgG1      TCS        $594.00 (paraffin sections)       

RDI-CD38abm-AT         AT13/5                         ---           mIgG1      purified    $594.00 (paraffin)

Purification Ammoniumsulphate precipitation followed by ionexchange chromatography and and affinity 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 CD38 antigen, which is strongly expressed on plasma cells. The CD38-antigen is also expressed on activated T-cells, terminally differentiated B-cells, early B- cells, monocytes, multiple myelomas, most cases of ALL (both T and B lineage), and some AML. The CD38-antigen is a single-chain Type II transmembrane protein.

Molecular mass 45kDa.

Application Identification of plasma cells.

Immunotyping of Leukeamias and Lymphomas.

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.

The expression is maximal on thrombin (1 U/ml) stimulated washed platelets.

FOR IN VITRO RESEARCH USE ONLY

Catalog#: RDI-CD38HIT2    $315.00/vial

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 IgG1k

CLONE: HIT2

Workshop Code: Workshop III, 155

Activity: Reacts with the 45kd tyep II single chain transmembrane glycoprotein present on thymocytes, activated T cells and terminally differentiated B cells (plasma cells). Other reactive cells include monocytes, macrophages, dendritic cells and some epithelial cells. The CD38 antigen acts as a NAD glycohydrolase in T lymphocytes.

Application: -Indirect immunofluorescence cell surface staining ,

-acetone fixed frozen tissue sections (with ABC detection, control tissue normal human thymus shows staining of thymocytes in cortex and medulla)

Titer for each application

References: 1) Br J. Haematol. 94:76 (1996)

                   2) Leucocyte Typing ZIII:Whie Cell Differentiation   Antigens. Oxford univ Press, New Yrk (1987)

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.

also available FITC conj: cat#RDI-CD38HIT2-FT    $438.00/100T

cat# RDI-CD38abm-290   $594.00

Presentation: 1ml lyophilized tissue culture supernatant containing 15mM sodium azide. Reconstitute with 1 ml of sterile distilled water , let set at least 30 minutes, gently mix before use.

Clone        SPC32

Ig Class    mIgG1

Antigen: Prokaryotic recombinant protein corresponding to the extracellular domain of the CD38 molecule.

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

Specificity: Human CD38 antigen (other species not tested)

Use: -frozen tissue with acetone fixation recommended.

        -formalin fixed paraffins ectiosn (with citrate buffer antigen release) approx 1:50-1:100 with 60 minutes primary antibody incubation at 25 o C. Standard ABC technique.

-not recommended for western blot

Positive ctrl: Tonsil

Staining: Membrane

Storage: Store unopened lyophilised antibody at 4 o C.Under these conditions,there is no significant loss in product performance up to the expiry date indicated onthe vial label.The reconstituted antibody is stable for at least two months when stored at 4 DEG C .For long term storage,it is recommended that aliquots of the antibody are frozen at -20 DEG C (frost-free freezers are not recommended).Repeated freezing and thawing must be avoided.Prepare working dilutions on the day of use.

For Research Use Only-not for use in or on humans or animals, not for use in diagnostics

Background: The CD38 molecule is a type II single transmembrane   glycoprotein with a molecular weight of 46kD.It is an ectoenzyme with the activities of ADP-ribosyl cyclase,cyclic ADP-ribose hydrolase,NAD glycohydrolase and is involved in both the formation and hydrolysis of cADPR,a second messenger that regulates the mobilisation of intracellular Ca 2+ ions.Although the CD38 molecule was originally identified as a T lymphocyte differentiation antigen,it is expressed in a wide range of cells and tissues.CD38 antigen can deliver potent growth and differentiation signals to lymphoid and myeloid cells.It is found on immature cells of the B and T cell lineages but not on most mature resting peripheral lymphocytes.It is also present on thymocytes,pre-B cells,germinal centre B cells,mitogen-activated T cells,Ig-secreting plasma cells, monocytes,NK cells,erythroid and myeloid progenitors in the bone marrow and brain cells. This Mab may be of value for phenotyping acute leukaemias and in research studies on the role of activated T cells in autoimmune and immunodeficiency diseases.It may also be used as a selective marker for germinal centre human B cells and for the detection of CD38 antigen in neurofibrillary tangles,the pathological indicator of Alzheimer ’s disease that occurs in the neuronal perikarya and proximal dendrites.

References

Deaglio S,Dianzani U,Horenstein A L,et al..Human CD38 ligand:a 120-KDA protein predominantly expressed on endothelial cells.The Journal of Immunology.156 :727-734 (1996).

Ellis J H,Barber K A,Tutt A,et al..Engineered anti-CD38 monoclonal antibodies for immunotherapy of multiple myeloma.The Journal of Immunology.155 :925-937 (1995).

Lund F,Solvason N,Grimaldi J C,et al..Murine CD38:an immunoregulatory ectoenzyme.Immunology Today.16(10):469-473 (1995).

Mizuguchi M,Otsuka N,Sato M,et al..Neuronal localization of CD38 antigen in the human brain.Brain Research.697 :235-240 (1995).

Malavasi F,Funaro A,Roggero S,et al..Human CD38:a glycoprotein in search of a function.Immunology Today.15(3):95-97 (1994).

also available clone AT13/5 (suitable for paraffin with EDTA buffer release)

cat#RDI-CD38abm-AT   $594.00/1ml

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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