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Goat Anti-Human Angiopoietin-4 (ANG-4)
Antigen Affinity Purified
(ANGPT4, AGP4, ANG-3, MGC138181)
Polyclonal Antibody
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Prod. No.:A368
Host:Goat
Pkg. Size:100 µg, 1 mg
Storage:-20°C to -70°C Detailed storage instructions below.
Description
Specificity:
Goat Anti-Human Angiopoietin-4 (ANG-4) recognizes Human ANG-4. 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:
Human angiopoietin-4 (Ang-4), alternatively named Ang-3, is a secreted glycoprotein belonging to the angiopoietin family. It has the characteristic structural motifs of angiopoietins including the coiled-coiled domain near the amino-terminus and a fibrinogen-like domain at the C-terminus. It shares 45%, 47% and 54% aa sequence identity with human Ang-1, human Ang-2 and mouse Ang-3, respectively. Although the sequence homology is much higher between the human and mouse counterparts for Ang-1 (97%) and Ang-2 (85%), mouse Ang-3 is believed to be an ortholog of human Ang-4 based on chromosomal localization studies. Human Ang-4 is highly expressed in lung and in cultured human umbilical vein endothelial cells (HUVECs). In contrast, mouse Ang-3 is expressed multiple mouse tissues. Human Ang-4 is an agonist that can bind and activate Tie-2, a receptor tyrosine kinase with immunoglobulin and epidermal growth factor homology domains expressed primarily on endothelial cells and early hematopoietic cells. Mouse Ang-3 has been reported to be a Tie-2 antagonist. It is likely that mouse Ang-3, like Ang-2, may exert agonist or antagonist activities depending on the cell context.

Known Species Cross Reactivity: Hu+
Immunogen
NS0-Derived Recombinant Human ANG-4
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 µl of sterile 1X PBS pH 7.2-7.3. This will result in a final antibody concentration of 0.1 mg/ml.
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, B
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-4 is approximately 1.2 ng/well.
IHC (NBF/Par.): This antibody should give satisfactory staining results when used at a concentration of 10 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-4 this polyclonal antibody can be used at a concentration of 0.1-0.2 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 detection limit for Human ANG-4 is 50 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.
Blocking: Approximately 1-3 μg/mL of this antibody will block 50% of 50 ng/mL of rhAng-4 to immobilized rhTie-2/Fc (100 μL of a 4 μg/mL solution was coated in each well) in a functional ELISA. At 50 μg/mL, this antibody will achieve approximately 85% blocking in the assay.
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 Angiopoietin-4 (Leinco Prod. No.: A353)
Anti-Human Angiopoietin-4–Biotin (Antigen Aff. Pur.) (Leinco Prod. No.: A360)
Anti-Human Angiopoietin-4 (Leinco Prod. No.: A384)
Products are for research use only. Not for use in diagnostic or therapeutic procedures.
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