RDI Divison of researchd 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.
CD19 Antibodies
Mouse anti-human CD19 antibodies
-4 clones CLB-B4/1,11G11 (purified, FITC & PE)
SJ25-C1 (purified, FITC & PE)
RFB-9 (purified)
BLY3 (purified)
CATALOG# CLONE# Workshop Host Form Price
RDI-M1345clb CLB-B4/1, 11G11 IV mIgG1 purified $375.00
RDI-M1436clb " " " " FITC $375.00
RDI-M1588clb " " " " PE $500.00
RDI-CBL141 SJ25-C1 III mIgG1 purified $375.00
RDI-CBL141FT " " " " FITC $375.00
RDI-CBL141PE " " " " PE $469.00
RDI-CBL495 RFB-9 -- mIgG1 purified $375.00 (Discontinued)
RDI-CBL506 BLY3 -- mIgM purified $375.00
(Cytolytic complement fixing clone)
Product Specification: mouse monoclonal anti-human CD19
PeliCluster CD19
CAT#RDI- M1345clb
Test/vial 200
Clone CLB-172 This clone has been derived from hybridization of SP2/0 cells with spleen cells of a BALB/c mouse immunized with spleen cells of a patient suffering from Hairy Cell Leukaemia This antibody has been clustered to CD19 in one of the international Workshop on Human White Cell differentiation Antigens.
Isotype Mouse, IgG1.
Source Hybridoma supernatant.
Purification 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 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 CD19- antigen (B4-antigen), which is expressed on human B lymphocytes. The monoclonal antibody is B lineage-specific and reacts with early B-cell precursors, pre-pre-B-cells, pre-B- cells, B-cells, intermediate B-cells, mature B-cells and some plasmacytoid cells. Plasma cells were found to be negative. The monoclonal antibody does not react with other haemopoïetic cells. The monoclonal antibody also reacts with pre-B-cell- lines, B lymphoblastoid cell-lines and Burkitt cell- lines, and with 50% of myeloma cell-lines. Virtually all non T-ALL, B-CLL and B-cell lymphomas were found to be positive, myeloma cells were found to be negative.
Molecular mass 95 kDa.
Methods Indirect immunofluorescence staining with analysis by flowcytometry or fluorescence microscopy.
References
1. Rie, M.A. de et al., J. of Immunol. Methods, 102, 187 (1987).
2. Rie, M.A. de Leukaemia Research, 12, 135 (1988).
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
mouse monoclonal CD19,conjugated to Fluorescein
PeliCluster CD19 F CAT#RDI- M1436clb
Test/vial 100
Clone CLB-172 This clone has been derived from hybridization of SP2/0 cells with spleen cells of a BALB/c mouse immunized with spleen cells of a patient suffering from Hairy Cell Leukaemia This antibody has been clustered to CD19 in one of the international Workshop on Human White Cell differentiation Antigens.
Isotype Mouse, IgG1.
Source Hybridoma supernatant.
Purification 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 and 10 mg BSA in PBS.
Preservative Mertiolate (0.001%).
Storage and stability Monoclonal antibodies should be stored 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 CD19- antigen (B4-antigen), which is expressed on human B lymphocytes. The monoclonal antibody is B lineage-specific and reacts with early B-cell precursors, pre-pre-B-cells, pre-B- cells, B-cells, intermediate B-cells, mature B-cells and some plasmacytoid cells. Plasma cells were found to be negative. The monoclonal antibody does not react with other haemopoïetic cells. The monoclonal antibody also reacts with pre-B-cell- lines, B lymphoblastoid cell-lines and Burkitt cell- lines, and with 50% of myeloma cell-lines. Virtually all non T-ALL, B-CLL and B-cell lymphomas were found to be positive, myeloma cells were found to be negative.
Molecular mass 95 kDa.
Application
Methods Direct immunofluorescence staining with analysis by flowcytometry or fluorescence microscopy.
References
1. Rie, M.A. de et al., J. of Immunol. Methods, 102, 187 (1987).
2. Rie, M.A. de Leukaemia Research, 12, 135 (1988).
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 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 VITRO RESEARCH USE ONLY
MOUSE ANTI-HUMAN CD19 + B LYMPHOCYTES
PRODUCT CODE :RDI-CBL141 $375.00/vial 200 Tests=200ug
Also available: FITC conjugated cat#RDI-CBL141FT $375.00/vial 100 Tests
PE conjugated cat#RDI-CBL141PE $469.00/vial 100 Tests
CLONE: SJ25-C1
ISOTYPE: IgG1
SOURCE: Mouse ascitic fluid.
PURIFICATION METHOD: Ammonium sulphate precipitation followed by DEAE ion exchange chromatography.
SPECIFICITY: 90-95 Kd MW lymphocyte surface antigen identified by monoclonal antibodies belonging to the CD19 cluster. Expressed from the earliest stages of B-progenitor development and on all peripheral B cells including germinal centre B cells, all B cell lines tested and B cell leukaemias tested. The antigen is lost on B cell maturation to plasma cells.
APPLICATIONS:
* Identification of CD19 positive B lymphocytes and malignant B cells by indirect immunofluorescence staining
* Leukaemia and lymphoma phenotyping
* Identification of normal and malignant CD19 positive B lymphocytes in cryostat sections of frozen tissue
* Isolation or removal of B lymphocytes by FACS or panning techniques
* Immunoprecipitation of the CD19 antigen
REFERENCES: Leucocyte Typing III (Oxford University Press) 1987, pp305-308
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 30 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
ANTI-HUMAN CD19 + B LYMPHOCYTES
PRODUCT CODE RDI-CBL495 $375.00/vial 200ug
Also available: FITC labeled cat#RDI-CBL495FT $375.00/vial 100 tests
PE labeled cat#RDI-CBL495PE $469.00/vial 100 tests
CLONE: RFB-9
ISOTYPE: IgG1
SOURCE: Tissue culture supernatant
PURIFICATION METHOD: Ammonium sulphate precipitation followed by DEAE ion exchange chromatography.
SPECIFICITY: 90-95 Kd MW lymphocyte surface antigen identified by monoclonal antibodies belonging to the CD19 cluster. Expressed from the earliest stages of B-progenitor development and on all peripheral B cells including germinal centre B cells, all B cell lines tested and B cell leukaemias tested. The antigen is lost on B cell maturation to plasma cells.
APPLICATIONS:
* Identification of CD19 positive B lymphocytes and malignant B cells by indirect immunofluorescence staining
* Leukaemia and lymphoma phenotyping
* Identification of normal and malignant CD19 positive B lymphocytes in cryostat sections of frozen tissue
* Isolation or removal of B lymphocytes by FACS or panning techniques * Immunoprecipitation of the CD19 antigen
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
MOUSE ANTI-HUMAN CD19 + B LYMPHOCYTES (CYTOLYTIC CLONE)
RODUCT CODE RDI-CBL506 $375.00/vial 200ug
Also available in BULK, without BSA and without azide CUSTOM QUOTES WELCOME
CLONE: BLY3
Isotype: IgM
PURIFICATION METHOD: Ammonium sulphate precipitation followed by DEAE ion exchange chromatography.
SPECIFICITY: The CD19 antigen (90kDa) is expressed from the earliest stage of B progenitor development, on all peripheral B cells including germinal centre B cells, and all B cell lines and B cell leukaemias tested. T cell and monocytic cell lines are negative and the antigen is lost on B cell maturation to plasma cells. The antigen is a type I integral membrane glycoprotein whose in vitro inhibition will influence B cell activation and proliferation.
APPLICATIONS: * Identification of CD19 positive B lymphocytes and malignant B cells by indirect immunofluorescence staining
* Leukaemia and lymphoma phenotyping
* Identification of normal and malignant CD19 positive B lymphocytes in cryostat sections of frozen tissue
* Isolation or removal of B lymphocytes by FACS or panning techniques * Immunoprecipitation of the CD19 antigen
* Lysis of CD19+ cells in the presence of complement (tested with rabbit complement)
- studies of CD19/Cd21 complex in B cell activation and the control of B cell development.
References: -Tedder, T.F et al. Immunology Today 15, 437-42 (1994)
-Tedder, T.F. & Isaacs, C.M.J. Immunol 143, 712-17 (1989)
-Kehrl, J. H. et al. Immunol Toady 15, 432-36 (1994)
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 IgM isotype monoclonal antibodies are aliquotted into small volumes and store at 4 DEG C or add an equal volume of glycerol and store at -20 DEG C. Dilute working solutions are best prepared on the day of use.