rev: August 20, 2007
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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).
Research Diagnostics Inc 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.
CD16 Antibodies
Mouse anti-human CD16 antibodies
-7 clones CLB-FcRgran/1,5D2 (purified ,FITC or PE)
CB16 (purified, FITC & PE)
GRM1 (purified, FITC or PE)
CLB-gran/11,5D7 (purified)
3G8 (please inquire for this clone, purified, FITC or F(ab'2) purified)
2H7 (paraffin sections)
CATALOG# CLONE# Wokshop Host Form Price
RDI-M1389clb CLB-FcRgran/1,5D2 IV & V mIgG2a purified $375.00
RDI-M1604clb " " " mIgG2a FITC $300.00
RDI-M1690clb " " " " PE $350.00
RDI-CBL541 GRM1 III mIgG2a purified $300.00
RDI-CBL541FT GRM1 III mIgG2a FITC $300.00
RDI-CBL541PE GRM1 III mIgG2a PE $350.00
RDI-M1390clb 16b CLB-gran/11,5D7 V mIgG2 purified $300.00
Granulocyte form only, GPI linked form FCgRIIIB
RDI-CD16abm-G8 3G8 - mIgG1 purified $250.00
RDI-CD16abm-GFT 3G8 - " FITC $300.00
RDI-CD16abm-GBT " - " Biotin $300.00
RDI-CD16abm-G2 " - " purified F(ab)'2 $1,000.00
RDI-CD16abm-2H7 2H7 -- mIgG2a TCS $450.00/1ml paraffin
Product Specification: murine monoclonal anti-human CD16
PeliCluster CD16
cat#RDI- M1389clb $375.00/vial
Test/vial 200
Clone CLB-FcR-gran/1, 5D2
This clone has been derived from hybridization of SP2/0 cells with spleen cells
of a BALB/c mouse immunized with human granulocytes. This antibody has been
clustered to CD16 in one of the international Workshop on Human White Cell
differentiation Antigens
Isotype Mouse IgG2a.
Source Ascites fluid of tumour
bearing BALB/c mice.
Purification Ammoniumsulphate
precipitation and ion exchange chromatography.
Packing Each vial contains 1 ml with approximately 0.1
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 CD16- antigen (the Fc gamma receptor
III), which is expressed on granulocytes, NK lymphocytes and macrophages (2,
4). The mobility of the CD16-antigen is dependent on the NA1/NA2 allotype of the neutrophil donor
(3). The monoclonal antibody inhibits the binding of human IgG to the Fc gamma receptor III (1, 4).
Molecular mass 45-72 kDa.
Application Enumeration of K -and NK-cell numbers in peripheral blood and
lymphoid tissue.
Methods Indirect immunofluorescence
staining with analysis by flowcytometry or
fluorescence microscopy.
References
1. Miedema F. et al., Eur.J. Immunol., 14, 518 (1984).
2. Tetteroo P.A.T. et al., Reinherz E.L. (editor) et al., Leucocyte
typing II,
3. Werner G. et al., Reinherz E.L. (editor) et al., Leucocyte typing II, Springer-Verlag, New York, 3, 109 (1986).
4. Tetteroo P.A.T. et al.,
Michael Mc A.J. (editor) et al., Leucocyte typing
III, Oxford University Press,
5. Huizinga T.W.J., Nature, 333, 667 (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
Product Specification: murine monoclonal CD16,conjugated to Fluorescein
PeliCluster CD16 F cat#RDI-M1604clb $300.00
Test/vial 100
Clone CLB-Fc-gran/1, 5D2
This clone has been derived from hybridization of
SP2/0 cells with spleen cells of a Balb/c mouse
immunized with human granulocytes. This antibody has been clustered to CD16 in
one of the international Workshop on Human White Cell differentiation Antigens.
Isotype The monoclonal isotype
is of the mouse subclass IgG2a.
Source Ascites fluid of tumour
bearing BALB/c mice.
Purification Ammoniumsulphate
precipitation and ion exchange chromatography.
Conjugation Conjugated with fluorescein thiocyanate isomer 1(FITC).
Molecular F/P ratio 5.5 - 9.5.
Packing Each vial contains 1 ml fluorescein
conjugated monoclonal antibody and 10 mg BSA in PBS.
Preservative Mertiolate (0.001%)
Storage 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 CD16- antigen (the Fc gamma Receptor
III), which is expressed on granulocytes, NK lymphocytes and macrophages (2,
4). The mobility of the CD16-antigen is dependent on the NA1/NA2 allotype of the neutrophil donor
(3). The monoclonal antibody inhibits the binding of human IgG to the Fc gamma Receptor III (1, 4)
Molecular mass 45 - 72 kDa
Application Enumeration of K- an NK-cell numbers in pheripheral
blood and lymfoid tissue.
Methods Direct immunofluorescence
staining with analysis by FACS or fluorescence microscopy.
References
1 Miedema, F et al., Eur. J. Immunol., 14, 518 (1984).
2 Tetteroo, P.A.T. et al., Reinherz, E.L. et al. (editor), Leucocyte typering II, Springr-Verlag, New York, 3, 27 (1986).
3 Werner, G. et al., Reinherz, E.L. et al. (editor), Leucocyrte typing II, Springer-Verlag, New York, 3, 109 (1986).
4 Tetteroo, P.A.T. et al., Mc Michael, A.J. et al., (editor), Leucocyte typing III, Oxford University Press, Oxford, 702 (1987).
5 Huizinga, T.W.J., Nature, 333, 667 (1988).
Application in direct Immunofluorescence techniques
Method with ficoll purified cells
Reagents and materials
- Fluorescein-conjugated monoclonal antibodies, diluted 1:2.5 in buffer
- Buffer: Phosphate Buffered Saline (PBS), containing 0.2% Bovine Serum Albumine (BSA)
- PFA: Para-Frmaldehyde 1%, pH 7.2,
containing 0.2% BSA
- Microtiter plates - 96 wells - V bottom or tybes.
Procedure
1 Prepare a mononucleair cell suspension with a concentratation of 5 x 10 4 cells/ml.
2 Ad 25 µl of cell suspension to microtiter wells or tubes
3. Add 25 µl of the diluted 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 granulocyte 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
- Fluorescein-conjugated monoclonal antibodies.
- Lysing solution (NH 4Cl), pH 7.2, freshly prepared from the stock solution.
- Buffer: Phosphate Buffered Saline (PBS), containing 0.2 % Bovine Serum Albumin (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 Analyze 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 - lynphocytes 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 Fluorescein-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
FOR IN VITRO RESEARCH USE ONLY
Product Specification: mouse monoclonal anti-human CD16b
PeliCluster CD16b CAT#RDI- M1390clb
Test/vial 200
Clone CLB-gran/11, 5D7
This clone has been derived from hybridization of
SP2/0 cells with spleen cells of a BALB/c mouse immunized with human
granulocytes. This antibody has been clustered to CD16b in the fifth
international Workshop on Human White Cell differentiation Antigens in
Isotype
Mouse IgG2a.
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 FcRIII receptor, (allo-antigen
NA1), which is expressed on human granulocytes and NA1-positive granulocytes.
Molecular mass 50-65 kDa.
Application NA1 phenotyping of
granulocytes.
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
MOUSE ANTI-HUMAN CD16 ANTIGEN FITC Conjugated
PRODUCT CODE :RDI-CBL525FT
$300.00/100T vial-
also available non labeled cat#RDI-CBL525 $300.00/200ug vial
PE labeled cat#RDI-CBL525PE $375.00/100 Tests
CLONE: CB16
ISOTYPE: IgG1
SOURCE: mouse ascitic fluid
PURIFICATION METHOD: Ion exchange chromatography.
SPECIFICITY: The CD16 molecule has been described as the low affinity Fc receptor (FcRIII) for complexed IgG which may exist either as a transmembranous form or as glycosyl
phosphatidylinositol form. It is expressed on NK
cells, granulocytes (PMN) and macrophages. The GPI-linked form is also
expressed on neutrophils.
Antigen distribution: Granulocytes >98%
LGL cells >85%
Peripheral blood lymphocytes 12 ± 8%
Thymocytes < 1%
Monocytes < 1%
B-cells (CD20+) < 1%
T-cells (CD3+) < 1%
APPLICATIONS:
* Identification of CD16+ leucocytes by direct immunofluorescence staining of viable cell suspensions.
* Identification of CD16+ leucocytes present within cryostat sections of frozen tissue.
* Studies on NK-characterization and chronic myeloid leukaemia.
REFERENCES: Leucocyte Typing V,
PRESENTATION: Each monoclonal is delivered in a sealed vial at a concentration
of not less than 100 Tests in 1ml 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. Use approx 10ul of conjugate per 1
million cells.
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. Dilute working solutions are best
prepared on the day of use. DO NOT fREEZE CONJUGATED
ANTIBODY. Protect from Light.
For research use only. Not supplied for use in human
diagnostic or therapeutic procedures
ANTI-Human CD16
PRODUCT CODE :RDI-CBL541 $300.00/200ug vial
-also available FITC labeled cat#RDI-CBL541FT $300.00/100T vial
PE labeled cat#RDI-CBL541PE $350.00/100 Tests
CLONE: GRM1
ISOTYPE: IgG2a
SOURCE: tissue culture supernatant
PURIFICATION METHOD: Protein A affinity
chromatography.
SPECIFICITY: The CD16 molecule has been described as the low affinity Fc receptor (FcRIII) for complexed IgG which may exist either as a transmembranous form or as glycosyl
phosphatidylinositol form. It is expressed on NK
cells, granulocytes (PMN) and macrophages. The GPI-linked form is also
expressed on neutrophils.
Antigen distribution: Granulocytes >98%
LGL cells >85%
Peripheral blood lymphocytes 12 ± 8%
Thymocytes < 1%
Monocytes < 1%
B-cells (CD20+) < 1%
T-cells (CD3+) < 1%
APPLICATIONS: * Identification of CD16+ leucocytes by direct immunofluorescence staining of viable cell suspensions.
* Identification of CD16+ leucocytes present within cryostat sections of frozen tissue.
* Studies on NK-characterization and chronic myeloid leukaemia.
-western blotting
REFERENCES: Leucocyte Typing V,
PRESENTATION: Each monoclonal is delivered in a sealed vial at a concentration of 200ug in 2ml 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 aliquoted and stored at 4 DEG C (up to one month) and then stored at -20 DEG C (long term). Avoid frequent freeze/thaw cycles. 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 (no BSA) cat#RDI-CBL541-1X $900.0/1mg, or NO BSA/NO AZIDE (cat#RDI-CBL541-1XP)
-Larger sizes upon request
DATA SHEET : Anti-human CD16 Monoclonal Antibody (Paraffin Sections/Western blot)
cat# RDI-CD16abm-2H7 $300.00/vial
Specificity: Human CD16 antigen
Clone: 2H7
Ig Class: IgG2a
Antigen: prokaryotic recombinant protein corresponding to the external domain
of the CD16 molecule common to both the transmembrane
form and the GPI-linked form.
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 suitable
Protocol: Immunohistochemistry:Typical working
dilution 1:20-1:40 after high temperature antigen unmasking technique using
0.01M Citrate pH6.0. 60 Minutes incubation at 25'C. Standard ABC technique.
Western blotting: 1:25-1:50 with chemiluminescence detection
Positive controls: tonsil
Staining pattern: Membrane staining of macrophages, neutrophils,
NK cells and granulocytes
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.
ref: -APMIS 101:319-329 (1993)
-Journal of Pathology. 172:189-197 (1994)
Stricly for in vitro research use only-Not for use in
or on humans or animals-not for use in diagnostics
CD16 antigen has a molecular weight of 50 to 70 kd and is a low affinity Fc
receptor for complexed IgG-Fc
gamma RIII expressed on natural killer (NK) clels,
granulocytes, activated macrophages and a subset of T cells expressing
alpha-beta or gamma-delta T cell antigen receptors. The CD16 anitgen exists both as a glycosyl-
phosphatidylinositol (GPI) anchored protein in polymorphonuclear cells and as a transmembrane
achored protein in NK cells.
Sample High Temperature Unmasking Technqiue for Paraffin Sections: 0.01M sodium Citrate Buffer (pH 6.0)
1) Cut and mount sections on slides coated with "apes"
2) Deparrafinize sections and rehydrate
to distilled water
3) Bring *or 0.01M sodium citrate buffer (pH 6.0) to a boil in a
Prestige/Presto stainless steel pressure cooker, using a hot plate. Cover but
do not lock lid.
4) Position slides into metal staining racks and lower into pressure cooker
ensuring slides are well immersed in buffer Lock the lid. The small valve will
rise.
5) When the pressure indicator valve (the large one) has risen after about 4-5
minutes, Incubate sections for 1 minute.
6) Remove pressure cooker from heat source and run under cold water with lid
on. When the small valve sinks open lid and remove slides and place immediately
into distilled water. DO NOT OPEN LID UNTIL THE SMALL VALVE Sinks or Vent Cover
Lock Drops (see manual with your particular unit).
7) Wash sections PBS buffer (10mM phosphate, 0.15M NaCl
pH 7.5) for 1 X 5 minutes (or 100mM TRIS, 0.15M NaCl,
pH 7.5)
8) Place sections in 1.5% hydrogen peroxide/methanol for 10 minutes (or use
other appropriate peroxidase blocking procedure).
9) Wash sections in distilled water for 2 X 5 minutes, then
wash sections in PBS buffer for 2 X 5 minutes.
10) Place sections in normal serum for 20 minutes.
11) Cover sections with primary antibody. (Optimum dilution, incubation time
and temperature must be determined for each antibody and in each lab).
12)
13) Incubate sections in secondary antibody for 30 minutes
14) Wash in PBS buffer for 2 X 5 minutes
15) Incubate slides in ABC complex for 30 minutes
16) Wash in PBS buffer for 2 X 5 minutes
17) Incubate slides in DAB or other suitable peroxidase
substrate
18) Wash in distilled water for 2 X 5 minutes
19) Counterstain with haematoxylin
(if required), dehydrate, coverslip, and mount
Material:
APES (3-aminopropyltriethoxysilane) cat#A3648 Sigma
To avoid sections becoming detached, sections should be mounted on "APES" covered slides, then dried at 37 DEG C overnight followed by drying at 56 DEG C for 60 minutes.
0.01M Sodium Citrate Buffer (ph 6.0): Add 2.94g of tri-sodium citrate to 1 liter of distilled water. Adjust pH 6 using 1.0M HCL.
Tris: 10mM Tris, 0.15M NaCl, pH 7.5.
PBS: 10mM Phosphate, 0.15M NaCl pH 7.5
Monoclonal anti-human CD16 (Fc gamma RIII)F(ab')2
cat# RDI-CD16abm-G2 $800.00/100ug
Presentation: 100ug in 0.ml ( mg/ml see spec sheet
with shipment) 50 mM Sodium Phosphate pH 7.5, 100 mM Potassium Chloride, 150mM NaCl.
Product was sterile filtered and vialed under aseptic
conditions. No preservative added.
Clone: 3G8
Isotype: Mouse IgG1
Immunogen: Human polymorphonuclear
leukocytes
PRODUCTION: Antibody was Protein A purified from (low FBS containing) tissue culture
supernatant. Immunoglobulin was enzymaticly cleaved
using Immobilized Ficin. F(ab')2 was separated from intact antibody and Fc fragments by protein A and Size exclusion
chromatography. Purity was >90% F(ab')2 by SDS-PAGE.
INFORMATION: Human CD16 is a low affinity receptor for complexed
IgG and occurs in two distinct forms, as a glycosyl phosphtidyl inositol (GPI)
anchored protein in granulocytes, and as an integral membrane protein on NK
cells and macrophages. Antibody 3G8 recognizes both allelic forms of CD16.
Antibody 3G8 blocks binding of complexed IgG to CD16.
References: H.B. Fleit, et al, (1982) Proc Natl Acad Sci
STORAGE: Store at 2 - 5oC. Open under aseptic conditions. Freeze/Thawing is not recommended.
PERFORMANCE: Five x 105 ficoll prepared human peripheral blood lymphocytes were washed and pre incubated ~5 minutes with 20 ul of 250 ug/ml human Ig (To block non specific binding) after which they were incubated 45 minutes on ice with 80 ul of anti-CD16 F(ab')2 at 5 ug/ml. Cells were washed twice and incubated with 2o reagent Goat anti-Mouse IgG/FITC , after which they were washed three times, fixed and analyzed using a BD FACS Calibur. A net 8.3% sub population of the cells stained positive with a mean shift of 1.72 log10 fluorescent units when compared to a Mouse IgG1 negative control at a similar concentration.
For Research Use Only
Also available: (intact)
non conjugated cat#RDI-CD16abm-G8 $250.00/100ug
FITC conjugated: cat#RDI-CD16abm-GFT $300.00/120T
Biotin conjugated: cat#RDI-CD16abm-GBT $300.00/100ug
PE conjugated: cat#RDI-CD16abm-GPE $400.00/100T
BULK QUOTES ON REQUEST
Research Diagnostics Inc
phone (800) 631-9384 or (973) 584-7093
fax (973) 584-0210
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