rev: July 26, 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.
CD20
Antibodies
Mouse anti-human CD20 antibodies
-5clones
CATALOG# CLONE# Workshop Host Form Price
RDI-M1644clb IH4 -- mIgG1 purified $375.00
RDI-M1646clb " " " " FITC $375.00
RDI-M1710clb " " -- " PE $438.00
RDI-CD20abm-2HPE 2H7 -- mIgG2b PE $500.00/102T
RDI-CD20abm-2HFT 2H7 -- " FITC $375.00/120T
RDI-CBL456 MEM 97 V mIgG1 purified $438.00
RDI-CBL161 (CD20/c) BC-1 + III mIgM purified $375.00
RDI-CBL161FT " " " " FITC $375.00
RDI-CBL143 B9E9 -- mIgG2a purified $375.00
RDI-CBL143FT " -- " FITC $375.00
RDI-CBL143PE " -- " PE $438.00
RDI-CD20abm-7D1 7D1 -- mIgG1 TCS $469.00 (paraffin & Wb)
RDI-CD20abm-L26 L26 -- mIgG1 TCS $469.00 (paraffin & WB)
RDI-CD20abm-MJ1 MJ1 -- mIgG1 TCS $469.00 (paraffin )
Product Specification: Mouse anti-human CD20
PeliCluster CD20
CAT#RDI-M1644clb $375.00/vial
Test/vial 200
Clone NKI-IH4
This clone has been derived from hybridization of SP2/0 cells with spleen cells of a BALB/c mouse immunized with mouse fibroblast cells transfected with plasmid DNA for CD20. This antibody has been clustered to CD20 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 affinity chromatography.
Packing Each vial contains 1 ml with approximately 0.05 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 CD20- antigen,
which is expressed on human B lymphocytes. Normal T-cells, polymorph nuclear
cells, monocytes and platelets were found to be negative.
Molecular mass 35-37 kDa.
Application Enumeration of B lymphocytes and their precursors in periphal
blood and lymphoid tissue.
Characterization of non-T (common) Acute Lymphoblastic Leukaemias.
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.
- microwell 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 microwell wells or tubes.
4 Add 5 µl monoclonal antibody to the microwell wells or tubes and mix gently.
5 Incubate for 30 minutes at 2-8 C.
6 Add 100 µl buffer to the microwell 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 microwell 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 CD20 F
CAT#RDI-M1646clb
Test/vial 100
Clone NKI-IH4
This clone has been derived from hybridization of SP2/0 cells with spleen
cells of a BALB/c mouse immunized with mouse fibroblast cells transfected
with plasmid DNA for CD20. This antibody has been clustered to CD20 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 affinity 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 CD20- antigen,
which is expressed on human B lymphocytes. Normal T-cells, polymorph nuclear
cells, monocytes and platelets were found to be negative.
Molecular mass 35-37 kDa.
Application Enumeration of B lymphocytes and their precursors in periphal
blood and lymphoid tissue.
Characterization of non-T (common) Acute Lymphoblastic Leukaemias..
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
- microwell 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 microwell wells or tubes.
3 Add 10 µl of the antibody to the microwell wells or tubes and mix gently.
4 Incubate for 30 minutes at 2-8°C.
5 Add 150 µl buffer to the microwell 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 microwell 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 microwell 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 lymphocyte 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
FOR IN RESEARCH VITRO USE ONLY
PRODUCT CODE :RDI-CBL161 $375.00/vial
also available: FITC labeled, cat#RDI-CBL161FT $375.00/vial 100T
CLONE: B - C1
ISOTYPE: IgM
SOURCE: Mouse ascitic fluid
PURIFICATION METHOD: Ammonium sulphate precipitation followed by gel
filtration.
SPECIFICITY: 37/35 kDa molecular weight membrane embedded non-glycosylated
phosphoprotein made up of two polypeptide chains which is expressed on B
cells.
APPLICATIONS:
* Studies in the regulation of B cell activation.
* Leukaemia and lymphoma phenotyping.
REFERENCES: Leucocyte Typing III, Oxford University Press. (1987)
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
PRODUCT CODE :RDI-CBL456 $438.00/vial
CLONE: MEM 97
ISOTYPE: IgG1
SOURCE: Mouse ascitic fluid.
PURIFICATION METHOD: Ion exchange chromatography following ammonium sulphate
precipitation.
SPECIFICITY: 37/35kDa molecular weight membrane embedded non-glycosylated
phospho-protein made up of two polypeptide chains and expressed on pan
B-cells.
APPLICATIONS:
* Studies in activation and regulatory signals in T & B-cells
* Studies in the regulation of B cell activation.
* Leukaemia and lymphoma phenotyping.
PRESENTATION: Each monoclonal is delivered in a sealed vial at a concentration
of 200µg/2ml in phosphate buffered saline solution containing 10mM sodium
azide. 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
PRODUCT CODE :RDI-CBL143 $375.00/vial 200ug
-also available FITC labeled cat#RDI-CBL143FT $375.00/vial 100T
PE labeled cat#RDI-CBL143PE $469.00/100TCLONE: B9E9
ISOTYPE: IgG2a
SOURCE: Tissue Culture superntanat
Immunizing Anitgen: Daudi cell line
PURIFICATION METHOD: Protein A affinity chromatography.
SPECIFICITY: 37/35 kDa molecular weight membrane embedded non-glycosylated
phosphoprotein with four membrane spanning domains as well as serine and
threonine rich cytoplasmic N-terminal and C-terminal domains. Only a minor
portion of the molecule, consisiting of one short and one longer loop is
exposed on the cell surface. CD20 is expressed on pre-B cells, resting activated
and neoplastic B cells but is absent from pro-B cells, plasma cells or mycioma
cells. It is also expressed on some non T-ALL cells and possibly on follicular
dendritic cells but not on monocytes, macrophages, neutrophils, red blood
cells or platelets. In normal lymph nodes and tonsils, CD20 is detected on
B cells in germinal centers while lower density expression is seen on cells
in the mantle zone. CD20 is associated with transmembrane CA 2+ transport
and is involved in the regulation of B cell proliferation and
differentiation.
APPLICATIONS: * Studies in flow cytometry of CD20 and immunocytochemistry
on frozen tissue sections.
REFERENCES: Leucocyte Typing V, Oxford University Press. (1995)
Tedder, T. et al. J. immunol 141, 4388 (1988)
Tedder, T. et al. Proc Natld Acad Sci 85, 208 (1988)
Tedder,,T. et al. J. Immunol 142, 2560 (1989)
Ledbetter, J et al, Human Immunology 15, 30 (1986)
Tedder, T. et al, Immunology Today 15, 450 (1994)
Landay, A et al AIDS 4, 479 (1990)
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 -200C. 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
BULK material (NO BSA) cat#RDI-CBL143-1X $1062.00/mg
NO BSA/NO AZIDE cat#RDI-CBL143-1XP $1062.00/1mg
cat# RDI-CD20abm-MJ1 $469.00/vial
Background: CD 20 is expressed in almost all lymphoid cells of B lineage
and has become the most widely used phenotypic marker for typing of
malignant lymphomas and other lymphoproliferative disorders in paraffin
sections. It is important to use a panel of antibodies when attempting
to type lymphomas
Specificity: Human CD20. Reactive with a cytoplasmic epitope localized on
the transmembrane protein (CD20). This 33 kD antigen is present on most B
lymphocytes.
Clone: MJ1
Ig Class: IgG1
Antigen: c terminal intracellular domain of a prokaryotic fusion protein.
Hybridoma partner: Mouse myeloma (p3-NS1-Ag4-1).
Preparation: Lyophilized tissue culture supernatant containing 15mM sodium
azide. Reconstitute with 1ml of sterile DI water.
USE: Frozen sections, and paraffin sections (with high temp unmasking
technique-especially required for tissue decalcified with acidic chelating
agents).
Protocol: Immunohistochemistry:Typical working dilution 1:25-1:50. High temperature antigen unmasking technique using 0.01M Na Citrate. 60 Minutes incubation at 25'C. Standard ABC technique.
Western blotting: must be determined (recommend clone 7D1 cat#RDI-CD20abm-7D1
$469.00/1ml (salso for paraffin sections & western blot)
Positive controls: human Tonsil, B cell immunoblastic and lymphoblastic
lymphomas.
Staining pattern: B lymphocyte membrane associated staining centers and mantle
zone areas of tonsil. Also, occasional B lymphocyte staining in interfollicular
regions and some B cell lymphomas.
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 freeze freezer, Avoid repeated freeze thaw cycles. Prepare fresh working dilutions daily.
Refs: Clin. Exp Immunol, 58:183-192 (1984)
American J. Pathology, 129:54-63 (1987)
Modern Pathology. 1:274-278(1988)
Strictly for in vitro research use only-Not for use in or on humans or
animals-not for use in diagnostics
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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