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.
CD31 Antibodies
Mouse anti-human CD31 antibodies
-4 clones
CATALOG# CLONE# Workshop Host Form Price
RDI-M1536clb CLB-HEC/75 IV mIgG1 purified $375.00
RDI-M1670clb " " " " FITC $375.00
RDI-CD31-HEC7 HEC7 VI mIgG2a purified $562.00 (blocking)
RDI-CBL473 WM59 IV mIgG1 purified $438.00
RDI-CD31abm-B3 158-2B3 -- mIgG1 see below forms available
RDI-CBL468 HC1/6 V mIgG1 purified $375.00 (paraffin)
RDI-CBL468FT " " " " FITC $375.00
RDI-CBL468PE " " " " PE $469.00
RDI-CD31abm-1A 1A10 -- mIgG1 TCS $500.00 (paraffin sections )
Mouse anti- human CD31(PECAM-1)/R-PE* clone 158-2B3
cat# RDI-CD31abm-BPE $500.00/120T
non conjugated: RDI-CD31abm-B3 $312.00/100ug
FITC conjugated: RDI-CD31abm-BFT $375.00/120T
Biotin conjugated: RDI-CD31abm-BBT $375.00/100ug
Package Size: 120T in 0.2ml 50mM sodium phosphate pH 7.5, 500mM Potassium Chloride, 150mM NaCL, 15% glycerol, 0.2% BSA and 0.04% NaN3.
Species: mouse IgG1
mAb name/Clone: 158-2B3
Immunogen: Stimulated human leukocytes
PRODUCTION: Protein A purified antibody from tissue culture supernatant was conjugated to R-Phycoerythrin through a sulfo-ester linkage. Unconjugated antibody was removed using size exclusion chromatography. Antibody conjugate is at 0.5mg/ml with an A565/A280 ratio of 3.19 (see batc sheet for bacth specific data)
WORKING DILUTION: 1:50 (80ul/test)
INFORMATION: Human CD31 is an adhesion molecule expressed on platelets, endothelial cells, leukocytes and their bone marrow precursors. CD31 plays a role in homophilic adhesion and heterophilic transendothelial migration (3).Antibody 158-2B3 reacts with domain 1 of CD31 and blocks homophilic interaction and heterophilic transendothelial migration (1).
Reference: 1) Leukocyte Typing VI (T. Kishimoto, et al, eds.) Garland Publishing, Inc., New York (1997) p. 362-372. 2) P.J. Newman, (1997) J Clin Invest 99: 3-10. 3) K.L. Yong, et al, (1998) Blood 91: 1196-1205.
STORAGE : Store at 2 - 5oC. Do not freeze! Protect from light.
PERFORMANCE: 500K cultured Jurkat cells were washed and incubated 45 minutes on ice with 80 ul of anti-CD31/R-PE at a 1:50 dilution (10 ug/ml). Cells were washed three times, fixed and analyzed using a BD FACS Calibur. Cells stained continued on back side positive with a mean shift of 1.87 log10 fluorescent units when compared to a Mouse IgG1/R-PE negative control (Catalog #RDI- MIGG1XPE $188.00) at a similar concentration. Binding was 100% blocked when pre incubated 10 minutes with 20 ml of 0.5 mg/ml anti-CD31 antibody (Catalog RDI-CD31abm-B3).
R-PE is covered under patents US 4,520,110; European 76,695 and Canadian 1,179.942.
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.
For Research Use Only
Product Specification: mouse monoclonal CD31
PeliCluster CD31
CAT#RDI- M1536clb $375.00/vial
Test/vial 200
Clone CLB-HEC/75 This clone has been derived from hybridization of SP2/0 cells with spleen cells of a BALB/c mouse immunized with human endothelial cells. This antibody has been clustered to CD31 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 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 CD31- antigen (gpIIa', PECAM-1), which is expressed on human thrombocytes. The monoclonal antibody reacts with thrombocytes, monocytes/macrophages, granulocytes and B-cells. In immunohistology the monoclonal antibody reacts with endothelial cells histiocytes (weak) and glomeruli.
Molecular mass 140 kDa.
Application Functional studies on cells.
Methods Indirect immunofluorescence staining with analysis by flowcytometry or fluorescence microscopy.
References
1. Mourik, J.A. et al., J. Biol. Chem., 260, 11300 (1985).
2. Newman, P.J. et al., Science, 247, 1219 (1990).
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
Product Specification: mouse monoclonal CD31,conjugated to Fluorescein
PeliCluster CD31 F
CAT#RDI- M1670clb
Test/vial 100
Clone CLB-HEC/75
This clone has been derived from hybridization of SP2/0 cells with spleen cells of a BALB/c mouse immunized with human endothelial cells. This antibody has been clustered to CD31 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 Affinity chromatography.
Conjugation Conjugated with fluorescein iso thiocyanate isomer 1 (FITC).
Molecular F/P ratio between 6.0 - 10.0.
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 CD31- antigen (gpIIa', PECAM-1), which is expressed on human thrombocytes. The monoclonal antibody reacts with thrombocytes, monocytes/macrophages, granulocytes and B-cells. In immunohistology the monoclonal antibody reacts with endothelial cells histiocytes (weak) and glomeruli.
Molecular mass 140 kDa.
Application Functional studies on cells.
Methods Direct immunofluorescence staining with analysis by flowcytometry or fluorescence microscopy.
References
1. Mourik, J.A. et al., J. Biol. Chem., 260, 11300 (1985).
2. Newman, P.J. et al., Science, 247, 1219 (1990).
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 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
FOR IN VITRO RESEARCH USE ONLY
DATA SHEET: mouse anti-human CD 31 (PECAM-1)
cat#RDI-CD31HEC7 $562.00/vial
Package Size: 200ug purified, sterile filtered, preservative, carrier free in 200ul PBS buffer
Species: mouse IgG2a
CLONE: HEC7
Activity: Reacts with human CD31 (PECAM-1)
Application: 1) Indirect immunofluorescence cell surface staining.
2) Blocking
3) immunohistochemistry
4) immunoprecipitation
Storage: Store at 4 DEG C. Avoid frequent freeze thaw cycles. Maximum long term stability with aliquoting and storage at -20 DEG C.
Precautions: For In vitro research Use Only. Not for use in or on humans or animals or for diagnostics.
References: -J. Immunol 1996, 156:326-335
-J. EXP MED ,AUGUST 1989,170:399-414
-Leucocyte Typing VI, Garland -Publishing, NY, 1997 pg 370-372
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.
MOUSE ANTI-HUMAN CD31 ANTIGEN (PECAM-1)
PRODUCT CODE :RDI-CBL 473 $438.00/vial 200ug
CLONE WM59
ISOTYPE IgG1
IMMUNIZING ANTIGEN Human myeloid leukaemic cells
FUSION PARTNER NS1 myeloma cell line
SOURCE Mouse ascitic fluid
PURIFICATION METHOD Ammonium sulphate precipitation followed by hydrophobic interaction chromatography
SPECIFICITY :The antibody reacts with a 130-140 kDa transmembranous glycoprotein that is expressed widely amongst cells of the vascular compartment. CD31 is expressed in platelets, cultured endothelial cells, monocytes, normal tissue macrophages in tonsil, lung and kidney as well as in skin biopsies from conditions such as sarcoidosis and lepromatous leprosy.
APPLICATIONS
*This antibody reacts with the platelet gpIIa molecule (also called PECAM-1) which is also expressed on endothelial cells
*Suitable for immunoprecipitation studies
*Suitable for flow cytometry by indirect labelling
REFERENCES
Leucocyte Typing IV, Oxford University Press (1989)
Leucocyte Typing V, Oxford University Press (1995)
DeLisser, H. M. et al. Immunol. Today 15, 490-95 (1994)
Newman, P. J. et al. Science 247, 1219-22 (1990)
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.
For Research Use Only. Not supplied for use in diagnostic or therapeutic procedures.
Mouse ANTI-HUMAN CD31 ANTIGEN
PRODUCT CODE :RDI-CBL468 $375.00/vial 200ug
Also available FITC labeled: cat#RDI-CBL468FT $375.00/100 Tests
PE labeled: cat#RDI-CBL468PE $469.00/100 Test
CLONE HC1/6
ISOTYPE IgG1
IMMUNIZING ANTIGEN PMA - treated U937 cells
FUSION PARTNER P3-x63Ag8.653 myeloma cell line
SOURCE Tissue culture supernatant
PURIFICATION METHOD Protein A affinity chromatography
SPECIFICITY The antibody reacts with a 130-140 kDa transmembranous glycoprotein that is expressed widely amongst cells of the vascular compartment. CD31 is expressed on platelets, cultured endothelial cells, monocytes, normal tissue macrophages in tonsil, lung and kidney as well as in skin biopsies from conditions such as sarcoidosis and lepromatous leprosy.
APPLICATIONS * Studies of the role of CD31 in leucocyte migration into inflammatory sites and in the process of angiogenesis* This antibody will stain frozen tissue sections* Suitable for immunoprecipitation studies* Suitable for flow cytometry either indirectly or directly when labelled with FITC or R-PE* Suitable for use on formalin fixed paraffin embedded sections after protease pre-treatment
REFERENCES Carbanas, C. et al. Eur. J. Immunol 19, 1373-1378 (1989)Leucocyte Typing V, Oxford University Press (1995)Stockinger, H. et al. J. Immunol. 145, 3889-97 (1990)DeLisser, H. M. et al. Immunol Today 15, 490-95 (1994)
LOT SPECIFIC DATA
PRESENTATION The monoclonal is presented at a concentration of 200µg/2ml in phosphate buffered saline containing 10mM sodium azide. 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.
-Bulk quotes on request-
DATA SHEET :Anti-human CD31 (PECAM-1) Monoclonal Antibody
(Paraffin Sections)
cat# RDI-CD31abm-1A $500.00/vial
Specificity: Human CD31
Clone: 1A10
Ig Class: IgG1
Antigen: prokaryotic recombinnat fusion protein corresponding to a portion of the extracellular domain downstream of the signal sequence of the CD31 (PEACAM-1) molecule
Hybridoma partner: Mouse myeloma (p3-NS1-Ag4-1).
Preparation: Lyophilized tissue culture supernatant containing 15mM sodium azide. Reconstitute with 1ml of sterile distilled water.
USE: Effective on paraffin wax embedded tissue using 0.01M citrate buffer solution combined with the high temperature antigen unmasking technique
frozen sections:Not effective
Protocol: Immunohistochemistry:Typical working dilution 1:50. High temperature antigen unmasking technique using 0.01M Citrate pH6.0. 60 Minutes incubation at 25'C. Standard ABC technique.
Positive controls: Breast carcinoma with vascular invasion
Staining pattern: Membrane
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 freze freezer, Avoid repeated freeze thaw cycles. Prepare fresh working dilutions daily.
Refs: Anatomic Pathology, 103 (4):443-448 (1995)
Stricly for in vitro research use only-Not for use in or on humans or animals-not for use in diagnostics