rev: March 9, 2004
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Anti-MOUSE
CD CLUSTERED
ANTIBODIES
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.
Anti-MOUSE CD119 Antibodies
see 2 monoclonals and polyclonals (also see below anti-mouse IFNGR beta chain)
see also anti-IFNGR
abs from the receptor data pack
cat# RDI-MCD119-GRBT 100ug Biotin labeled $344.00
Package Size: 100ug purified in 0.2ml (0.5mg/ml) buffer and 0.09% 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.)
Species: rat IgG2a (brown Norway)
CLONE: GR20
Immunogen: IFNG receptor affinity enriched from membranes of the
monomyelocytic cell line WEHI-3
Activity: Reacts with mouse CD119, the alpha subunit of the interferon gamma
receptor. This receptor is expressed by mouse leukocytes, endothelial and
epithelial cells but not mature erythrocytes. Clone GR20 Mab recognizes an
epitope susceptible to certain fixatives in the ligand-binding site of the
receptor, therefore it block binding of mouse IFN-G. It has been reported
that GR20 antibody can prevent the ability of mouse IFNG to prime macrophages
for tumor cell killing. IFNG has been reported to induce the expression of
LY-6A/E antigen on cell lines and normal B cells. It has been observed that
GR20 Mab inhibits IFN-G induced expression of Ly-6A/E on the mouse B lymphoma
line A20. Binding of the GR20 ab to CD119 is inhibited in the presence of
IFN-G since the GR20 antibody recognizes an epitpe in the ligand binding
site of the IFNGR. This antibody reportedly does not have any agonist
effect.
Application: -Indirect immunofluorescence cell surface staining (flow cytometry)
-immunoprecipitation (purified /non conjugated ab)
-western blot (non reducing conditions)
-immunocytochemistry
-ELISA
-purification of detergent solubilized receptor
References: 1) Proc Natl Acad Sci USA 85:6282-6286
2) J. Leukoc Biol 51:507-516
3) J Biol Chem 265:4064-4071
4)
J. Immunol 144:2197-2204
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.
CAT# RDI-PB32580 $375.00/vial
Description: Rabbit Anti-Mouse Interferon Gamma Receptor Chain 1,
non-neutralizing.
Quantity Shipped: 1 vial of 50 ug in 50 ul.
Concentration: 1 mg/ml.
Bioactivity: Binds to but does not neutralize mouse interferon gamma
receptor.
Usage: Western blot; immunoprecipitation; immunohistochemistry; flow cytometry.
Shipping Temperature: ice pack
Physical State of Product During Shipping: Frozen
Special Conditions/Comments: After receipt, this product should bekept at -20'C or below for retention of full activity. When thawing, the contents of the tube should be apportioned in separate tubes so that freezing and thawing is kept to a minimum.
-For In Vitro Research Use Only-
1 (RMMGR-1) non-neutralizing.
CAT# RDI-PB22580 $375.00/vial
Quantity Shipped: 1 vial of 50 ug in 50ul.
Concentration: 1.0 mg/ml.
Buffer: Phosphate buffered saline (PBS)
Isotype: To be determined.
Bioactivity: Binds to but does not neurtralize mouse interferon gamma receptor.
Usage: Western blot; immunoprecipitation; immunohistochemistry; flow cytometry.
Shipping Temperature: ice pack
Physical State of Product During Shipping: Frozen
Special Conditions/Comments: After receipt, this product should be kept at -20'C or below for retention of full activity. When thawing, the contents of the tube should be apportioned in separate tubes so that freezing and thawing is kept to a minimum.
-For In Vitro Research Use Only-
(CD119) MONOCLONAL ANTIBODY (Nonblocking Antibody)
cat# RDI-MCD119-2E2 $500.00
Presentation: 0.5mg in 1mg (0,5mg/ml) buffer with 0.09% sodium azide
Clone: 2E2
Immunogen: purified preparation of soluble recombinant mouse IFN-GRalpha chain protein.
Isotype: Armenian Hamster IgG
Purification: purified from tissue culture supernatant via Protein A affinity chromatography
Storage: The antibody solution should be stored undiluted at 4°C.
Use: -FACS (1ug/million cells):clone 2E2 MAB is a nonblocking antibody that can be used for the unobstructed immunofluorescent staining and flow cytometric analysis of cells in systems where the ligand (i.e., IFN-G ) for IFN-G receptors is present. A three-layer staining protocol is recommended for maximizing the detection IFN-GRa chains expressed by cells .
-IP: The 2E2 antibody has been reported to be useful for the immunoprecipitation of IFN-GRa chains from lysates of cloned mouse T cells.
Background: The 2E2 antibody recognizes the extracellular region of the 90 kDa alpha chain subunit of the mouse interferon- receptor (IFN- R ; aka, CD119). The functionally active-form of the mouse IFN- receptor consists of two (or more) subunits, with IFN- R responsible for IFN- binding and both the IFN- R and IFN- R chains required for the transduction of biologic responses. IFN- R is expressed by a variety of cell lines and normal mouse cells (except mature erythrocytes) including T cells, B cells, NK cells, monocytes, neutrophils, fibroblasts, epithelial and endothelial cells. The 2E2 antibody is a non-neutralizing antibody; it does not block the binding of IFN-G to its receptor.
For In Vitro Research Use Only-not responsible for patent infringement or other violations that may occur with the use of our products.
REFERENCES:
1. Bach, E. A., S. J. Szabo, A. S. Dighe, A. Ashkenazi, M. Aguet, K. M. Murphy, and R. D. Schreiber. 1995. Ligand-induced autoregulation of IFN-gamma receptor beta chain expression in T helper cell subsets. Science 270:1215-1218.
Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
see also hamster anti-mouse IFNGRB clone MOB-47 cat#RDI-MIFNGRB-47 $500.00/0.5mg
Sample staining protocol:
BALB/c spleen cells (including erythrocytes) were preincubated (~15 min., 4°C) with purified 2.4G2 antibody [rat anti-mouse CD16 /CD32 (Fc II); 1 µg antibody/10 6 cells]. The cells were stained (30 min., 4°C) with purified 2E2 antibody (0.5 µg mAb/10 6 cells; After washing, the cells were incubated (30 min., 4°C) with a biotin-conjugated cocktail of mouse anti-hamster antibodies (Clones G70-204 + G94-56; Cat. No. RDI-12102D $375.00; 0.5 µg mAb cocktail/10 6 cells). Finally, the cells were washed and incubated with R-PE-conjugated streptavidin 0.015 µg PE-SA/10 6 cells) and FITC-anti-CD4 (clone RM4-5;) and FITC-anti- CD8 (clone 53-6.7; ), both at 0.06 µg mAb/10 6 cells. After washing, the cells were analyzed with a FACScanZ Flow Cytometer. The data indicates that mouse erythrocytes are uniformly negative for IFN-GR alpha expression whereas most T cells (i.e., CD4 + /CD8 + cells) and CD4-CD8-. lymphocytes constitutively express medium levels of IFN-GR alpah chains.
cat# RDI-MIFNGRB-47 $500.00
Presentation: 0.5mg in 1ml (0.5mg/ml) buffer with 0.09% sodium azide
Clone: MOB-47
Isotype: Armenian Hamster IgG
Immunogen: purified soluble extracellular domain of the mouse IFN-GR beta chain protein.
Purification: purified from tissue culture supernatant via Protein A affinity chromatography.
Storage: The antibody solution should be stored undiluted at 4°C.
Use: -FACS:1ug/million cells (1 µg antibody/10 6 cells) A three-layer staining protocol is recommended for maximizing the detection IFN- R chains expressed by cells
-IP/WB: The MOB-47 antibody has been reported to be useful for the immunoprecipitation and Western blotting of IFN-GR beta chains from lysates of cloned mouse T cells.
background: The MOB-47 antibody recognizes the 60-65 kD beta chain subunit of the mouse interferon- receptor (IFN- R ). The IFN- receptor beta chain is expressed by a variety of normal mouse cell types and cell lines that are responsive to IFN- including T and B lymphocytes, NK cells, monocytes, macrophages, granulocytes and fibroblasts. The IFN- R receptor epitope recognized by the MOB-47 antibody is blocked by bound IFN-G .
Note: The IFN-GRbeta receptor epitope recognized by the MOB-47 antibody is blocked by bound IFN-G . Thus, it has been reported that cells which have bound IFN- 1 can be pretreated with dilute acid to remove the bound ligand before they are stained.
REFERENCES:
1. Bach, E. A., S. J. Szabo, A. S. Dighe, A. Ashkenazi, M. Aguet, K. M. Murphy, and R. D. Schreiber. 1995. Ligand-induced autoregulation of IFN-gamma receptor beta chain expression in T helper cell subsets. Science 270:1215-1218.
Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
For Research Use only
sample staining protocol:
BALB/c spleen cells and A20 cells (mouse B cell lymphoma) were preincubated (~15 min., 4°C) with purified 2.4G2 antibody [rat anti-mouse CD16 1 µg antibody/10 6 cells] to block nonspecific staining due to immunoglobulin Fc receptors. The cells were stained (45 min.,4°C) with purified MOB-47 antibody (either at 0.5 µg mAb/10 6 BALB/c cells or at 0.125 µg mAb/10 6 A20 cells; . After washing, the cells were incubated (30 min., 4°C) with a biotin-conjugated cocktail of mouse anti-hamster antibodies (Clones G70-204 + G94-56; Cat. No.RDI-12102D; 0.5 µg $375.00 mAb cocktail/10 6 cells). Finally, the cells were washed and incubated with R-PE-conjugated streptavidin (0.015 µg PE-SA/10 6 cells). After washing, the cells were analyzed with a FACScanZ Flow Cytometer.
CHAIN 2 (Beta) (non neutralizing)
CAT# RDI-PB32585 $375.00
Quantity Shipped: 1 vial of 50 ug in 53 ul.
Concentration: 1 mg/ml.
Bioactivity: Binds to but does not neutralize mouse interferon gamma
receptor.
Usage: Western blot; immunoprecipitation; immunohistochemistry; flow cytometry
Shipping Temperature: ice packs
Physical State of Product During Shipping: Frozen
Special Conditions/Comments: After receipt, this product should be kept at -20'C or below for retention of full activity. When thawing, the contents of the tube should be apportioned in separate tubes so that freezing and thawing is kept to a minimum.
-For In Vitro Research Use Only-
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RDI Divison of researchd Industries Intl
San Jose, 95123 CA Snell ave 658
USA
or 408-780-0908
EMAIL:margaret@cellular-research.com