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Goat Anti-Human Angiogenin (ANG)
Antigen Affinity Purified
Polyclonal Antibody
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Prod. No.:A363
Host:Goat
Pkg. Size:100 µg, 1 mg
Storage:-20°C to -70°C Detailed storage instructions below.
Description
Specificity:
Goat Anti-Human Angiogenin (ANG) recognizes Human ANG. This antigen affinity purified polyclonal antibody was purified using a proprietary chromatographic technique that includes covalently immobilizing the antigen proteins or peptides to agarose based beads. This purification method enhances specificity, reduces nonspecific binding of extraneous IgG and provides you with the most reliable reagent available for your early discovery research.

Background:
Angiogenin (Ang) is a small polypeptide that is implicated in angiogenesis (formation of new blood vessels) in tumor growth . However, angiogenin is unique among the many proteins that are involved in angiogenesis in that it is also an enzyme with an amino acid sequence 33% identical to that of bovine pancreatic ribonuclease (RNase). Moreover, although Ang has the same general catalytic properties as RNase A - it cleaves preferentially on the 3' side of pyrimidines and follows a transphosphorylation/hydrolysis mechanism - its activity differs markedly both in magnitude and in specificity.

Although angiogenin contains counterparts for the key catalytic residues of bovine pancreatic RNase A, it cleaves standard RNase substrates 105 - 106 times less efficiently than does RNase A. Despite this apparent weakness, the enzymatic activity of Ang appears to be essential for biological activity: replacements of important active site residues invariably diminish ribonuclease and angiogenesis activities in parallel, and a substitution that increases enzymatic activity also enhances angiogenic potency.

Angiogenin may function as a tRNA-specific ribonuclease that binds to actin on the surface of endothelial cells; once bound, angiogenin is endocytosed and translocated to the nucleus, thereby promoting the endothelial invasiveness necessary for blood vessel formation. Angiogenin induces vascularization of normal and malignant tissues, and abolishes protein synthesis by specifically hydrolyzing cellular tRNAs.

Alternative splicing results in two transcript variants encoding the same protein. This gene and the gene that encodes ribonuclease, RNase A family, 4 share promoters and 5' exons. Each gene splices to a unique downstream exon that contains its complete coding region.

Immunogen
E. coli-Derived Human ANG
Formulation
This antigen affinity purified polyclonal 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
We suggest aseptically reconstituting this lyophilized, antigen affinity purified polyclonal antibody with 1.0 ml of sterile 1X PBS pH 7.2-7.3. This will result in a final antibody concentration of 0.1 mg/ml.
Endotoxin
<0.1 EU/µg as determined by the LAL method
Storage and Stability
The lyophilized antigen affinity purified polyclonal antibody can be stored desiccated at -20°C to -70°C for twelve months from date of receipt. 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
Indirect ELISA, IHC (NBF/Par.), WB
Indirect ELISA: This antibody can be used in an indirect detection ELISA at 0.5-1.0 µg/ml with a suitable second step reagent such as Donkey Anti-Goat IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: G505). The detection sensitivity of this indirect ELISA for Human ANG is approximately 0.06 ng/well.
IHC (NBF/Par.): This antibody should give satisfactory staining results when used at a concentration of 2-5 µg/ml. The recommended secondary antibody for IHC is Donkey Anti-Goat IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: G505). 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).
Western Blotting: To detect Human ANG this polyclonal antibody can be used at a concentration of 0.1 µg/ml. This polyclonal antibody should be used in conjunction with compatible second-step reagents such as Donkey Anti-Goat IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: G505) and a chromogenic substrate such as TMB HRPO Membrane/IHC Substrate UltraSensitive™ One Component "Ready Use" (Leinco Prod. No.: T343). The sensitivity of detection may increase up to 50 fold when a chemiluminescent substrate is used. A suitable Western blotting control is Recombinant Human Angiogenin (Leinco Prod. No.: A339).
Each investigator should determine their own optimal working dilution for specific applications.


Country of Origin
USA
Related Products
Donkey Anti-Goat IgG (H&L)-Horseradish Peroxidase (Leinco Prod. No.: G505)
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 Human Angiogenin (Leinco Prod. No.: A339)
Anti-Human Angiogenin (Leinco Prod. No.: A373)
Anti-Human Angiogenin-Biotin (Antigen Aff. Pur.) (Leinco Prod. No.: A396)
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