Product Datasheet
www.leinco.com
excellence in early discovery research       



Anti-Human Amphiregulin (AR)
Purified No Carrier Protein
(SDGF)
Monoclonal Antibody
Share |


Prod. No.:A372
Clone:31221
Isotype:Mouse IgG1
Pkg. Size:500 µg, 1 mg
Storage:-20°C to -70°C Detailed storage instructions below.
Description
Specificity:
Mouse Anti-Human Amphiregulin (AR) (Clone 31221) recognizes an epitope on Human AR. This monoclonal antibody was purified using multi-step affinity chromatography methods such as Protein A or G depending on the species and isotype.

Background:
Amphiregulin (AR) is a member of the EGF family of cytokines which is comprised of at least ten proteins including EGF, TGF-α, HB-EGF, and the various heregulins. All of these cytokines are synthesized as transmembrane precursors and are characterized by the presence of one or several EGF structural units in their extracellular domain. The soluble forms of these cytokines are released by proteolytic cleavage. Amphiregulin was originally isolated from the conditioned media of a PMA-treated MCF-7 human breast carcinoma cell line. The Amphiregulin mRNA expression can be detected in numerous carcinoma cell lines and the epithelial cells of various human tissues including colon, stomach, breast, ovary, kidney, etc. AR is also a growth inhibitor for various tumor cell lines. In certain colon carcinoma cell lines, AR has been shown to be an autocrine growth factor. Amphiregulin can bind to the EGF receptor. It has been suggested that in certain cell types, AR bioactivity may be mediated through the EGF receptor.

Known Species Cross Reactivity: Hu+
Host Species
Mouse
Immunogen
Purified Recombinant Human AR (>98%)
Formulation
This monoclonal antibody has been 0.2 µm filtered and lyophilized from modified Dulbecco's phosphate buffered saline (1X PBS) pH 7.2 - 7.3 containing 5.0% w/v trehalose with no calcium, magnesium or preservatives present.
Reconstitution
This lyophilized antibody may be aseptically reconstituted with sterile PBS to a final volume of 1 ml. This will result in a final antibody concentration of 500 µg/ml.
Endotoxin
<0.1 EU/µg as determined by the LAL method
Storage and Stability
The lyophilized antibody can be stored desiccated at -20°C to -70°C for up to twelve months. The reconstituted antibody can be stored for at least four weeks at 2-8°C. For long-term storage of the reconstituted antibody, aseptically aliquot into working volumes and store at -20°C to -70°C in a manual defrost freezer. Avoid repeated freeze thaw cycles. No detectable loss of activity was observed after six months.
Reported Applications
ELISA Cap, IHC (NBF/Par.), N, WB
ELISA Sandwich: This antibody is useful as the capture antibody in a sandwich ELISA. The suggested coating concentration is 2-8 µg/ml. A suitable detection antibody is Anti-Human Amphiregulin-Biotin (Antigen Aff. Pur.) (Leinco Prod. No.: A395) at a concentration of approximately 0.1-0.4 µg/ml. A suggested standard for this assay is Recombinant Human Amphiregulin (Leinco Prod. No.: A338).
IHC (NBF/Par.): This antibody should give satisfactory staining results when used at a concentration of 15 µg/ml. The recommended secondary antibody for IHC is Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M114). For chromogenic detection with high signal and low background use 3, 3'-Diaminobenzidine Tetrahydrochloride (DAB) Substrate Kit (Leinco Prod. No.: D100) or Optional Metal Enhanced DAB Substrate Kit (Leinco Prod. No.: K107).
Neutralization: This antibody is useful for neutralization of Human AR bioactivity. The antibody dose required to neutralize 50% (ND50) of the biological activity of Human AR (at 50 ng/ml) is 5-15 µg/ml.
Western Blotting: To detect Human AR this monoclonal antibody can be used at a concentration of 1-2 µg/ml. This monoclonal antibody should be used in conjunction with compatible second-step reagents such as Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M114) and a chromogenic substrate such as TMB HRPO Membrane/IHC Substrate UltraSensitive™ One Component "Ready Use" (Leinco Prod. No.: T343). The detection limit for Human AR is 5 ng/lane under either reducing or non-reducing conditions. The sensitivity of detection may increase up to 50 fold when a chemiluminescent substrate is used. A suitable Western blotting control is Recombinant Human Amphiregulin (Leinco Prod. No.: A338).
Each investigator should determine their own optimal working dilution for specific applications.


Country of Origin
USA
Related Products
Anti-Human Amphiregulin-Biotin (Antigen Aff. Pur.) (Leinco Prod. No.: A395)
Recombinant Human Amphiregulin (Leinco Prod. No.: A338)
Goat Anti-Mouse IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: M114)
3, 3'-Diaminobenzidine Tetrahydrochloride (DAB) Substrate Kit (Leinco Prod. No.: D100)
Optional Metal Enhanced DAB Substrate Kit (Leinco Prod. No.: K107)
TMB HRPO Membrane/IHC Substrate UltraSensitive™ One Component "Ready Use" (Leinco Prod. No.: T343)
Recombinant Mouse Amphiregulin (Leinco Prod. No.: A354)
Anti-Human Amphiregulin (Antigen Aff. Pur.) (Leinco Prod. No.: A362)
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
359 Consort Drive | St. Louis, MO 63011 | (800) 538-1145 | (636) 527-5545 fax
800.538.1145 ® Copyright Leinco Technologies, Inc. 2006 – 2012