Angiopoietin ANTISERA-Human IgG Subclasses Book Section 5.3 & 5.4

Angiopoietin ANTISERA

-Angiopoietin-1 is a secreted ligand for Tie-2, a cell membrane receptor. Preliminary biochemical analysis of Ang-1 reveal potential fibrinogen-like domain at the carboxy terminus and coiled-coil regions in the amino terminus. Ang-1 is an angiogenic factor that is thought to be involved in endothelial development. A related protein, angiopoietin-2 (ang-2), has been identifiec s a naturally occurring antagonist of Ang-1 avtivation of Tie-2. In adult tissue, Ang-2 expression seems to be restricted to sites of vascular remodeling.

ANG-1 is prominently expressed in the myocardium of atrium and ventricle, mesenchymal and smooth muscle cells surrounding most blood vessels, an dlung. ANG-2 expression is abundant in the dorsal aorta and major aortic branches. ANG-2 transcripts in fetal liver were restricted to cells at the lumen of hepatic vessels. In the adult, ANG-1 was also expressed in the heart and liver. In contrast, ANG-2 was found in tissues that may undergo vascular remodeling (ovary, placenta and uterus).

Expected size of Ang-1 is ~55 kDa (although glycosylated forms have been seen at approx ~ 70 kDa , glycosylated, secreted protein in the conditioned media from cells expressing recombinant Ang-1. ). The predicted size of Ang-2 is ~55 kDa. Like Ang-1, its size may higher due to glycosylation  see Maisonpierre C et al (1997) Science 277, 55.

 

The following antibodies are purified to homogeneity and supplied at 200 ug/ml in PBS with 0.2% gelatin and 0.1% azide


Rabbit anti-ANG-1 (Angiopoietin-1)  HUMAN/MOUSE/RAT

cat#RDI-ANGIOP1XabR   $400.00/100ug vial

-rabbit polyclonal antibody IgG, peptide affinity purified

-A 20 amino acid peptide sequence near the amino-terminus of the mouse Ang-1

-The 20 AA Ang-1 immunogenic peptide sequences is 90% (18/20 aa)

homologous with human Ang-1. Ang-1 control peptide has no significant sequence homology with Ang-2. Antibody crossreactivity in various species is not established

-specific for Angiopoietin-1

-mouse and human reactive

-suitable for western blotting (approx 1ug/ml with chemiluminescence detction ,band approx 55-70kda)

-not tested for histochemistry (would recommend acetone fixed frozens or PFA fixed sections, titer from 10ug/ml with ABC/DAB detection, positive control atrial tissue)

-control/blocking peptide: cat#RDI-ANGIOP1X-CX $150.00/100ug in 0.1ml PBS, pH 7.4 and 0.02% sodium merthiolate


Rabbit anti-ANG-2 (Angiopoietin-2) MOUSE (human?)

cat#RDI-ANGIOP2XabR   $400.00/100ug vial

-rabbit polyclonal antibody IgG, peptide affinity purified

-A 21 amino acid peptide sequence near the amino-terminus of the  mouse Ang-2 was used to generate anti- Ang-2. The peptide was coupled with KLH (carrier protein)

-The 21 AA Ang-2 immunogenic peptide sequences is 71% (15/21 aa) homologous with human Ang-2. Ang-2 control peptide has no substantial homology with Ang-1

-specific for Angiopoietin-2

-mouse and with less reactivty human reactive

-suitable for western blotting (approx 1ug/ml with chemiluminescence detction ,band approx 55kda)

-not tested for histochemistry (would recommend acetone fixed frozens or PFA fixed sections, titer from 10ug/ml with ABC/DAB detection, positive control dorsal aorta tissue

-control/blocking peptide: cat#RDI-ANGIOP2X-CX $150.00/100ug in 0.1ml PBS, pH 7.4 and 0.02% sodium merthiolate


Goat anti-human Angiopoietin-2

cat#RDI-ANG218NabG   $425.00/100ug

-Goat polyclonal antibody IgG, peptide affinity purified

-An 18 amino acid peptide sequence near the amino-terminus of the human Ang-2 was used to generate anti- Ang-2. The peptide was coupled with KLH (carrier protein)

-specific for Angiopoietin-2

-human reactive (other species not tested)

-suitable for western blotting (approx 1ug/ml with chemiluminescence detction ,band approx 55kda)

-not tested for histochemistry (would recommend acetone fixed frozens or PFA fixed sections, titer from 10ug/ml with ABC/DAB detection, positive control dorsal aorta tissue)

-control/blocking peptide: cat#RDI-ANG218N-CPX $150.00/100ug in 0.1ml PBS, pH 7.4 and 0.02% sodium merthiolate


Related Product(s):

RDI-MANGIOP1abrX     rabbit anti-angiopoietin 1 (mouse specific) $400.00/0.2ml

RDI-MANGIOP2ABRX  rabbit anti-Angiopoietin 2 (Mouse specific) (Rabbit) $400.00/0.2ml

RDI-MANGIOP1R-CP   CONTROL PEPTIDE For Anti-ANGIOPOIETIN 1 (Mouse) $150.00

RDI-MANGIOP2R-CP   CONTROL PEPTIDE For Anti-ANGIOPOIETIN 2 (Mouse) $150.00


Rabbit anti--Mouse Angiopoietin 1 NEW!

Code: RDI-MANGIOP1ABRX    $400.00/0.2ml

Presentation: 0.2ml neat rabbit sera in 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 with 0.01% sodium azide

Lot #: see sheet with shipment

Protein Concen: 85 mg/ml (by Refractometry)

Storage Conditions: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening.

Background Information: Angiopoeitin-1 (Ang-1) and Antiopoietin-2 (Ang2) are important for development of the endothelium, by regulating tyrosine phosphorylation of the membrane receptor Tie-2/Tek. Ang-1 binding to Tie-2/Tek causes phosphorylation of the receptor. Ang-2 competes for this binding, and thus blocks receptor phosphorylation. Ang-1 has potential fibrinogen-like domain at the carboxy terminus and coiled-coil regions in the amino terminus. Ang-1 is prominently expressed in the myocardium of atrium and ventricle, mesenchymal and smooth muscle cells surrounding most blood vessels, and lung. In the adult, Ang-1 is also expressed in the heart and liver. Anti-Ang 1 and anti-Ang 2 antibodies recognize a band above 55 kDa. Both antibodies show banding at 75 kDa which resembles the original size, because these are soluble highly glycosylated proteins, which should run higher that the calculated molecular weight of 55 kD.

Application Note(s): Suitable for western blotting and other antibody based assays. A 1:500 dilution is recommended for western blotting. The reaction of this antiserum directly with cell supernatants may result in high background due to reactivity of components in the serum. This can be alleviated by first immunoprecipitating the antibody:antigen complex and then detecting the antigen. This method results in very clean and strong signal.

Recommended Dilutions: This product was assayed by immunoblot against supernatants of angiopoietin-expressing endothelial cells from human and mouse sources. A single band of the expected apparent molecular weight was observed at a 1:500 dilution. Researchers should determine optimal titers for other applications.

Purity and Specificity: This antiserum is directed against mouse angiopoietin 1 and shows no reactivity with angiopoietin 2 on mouse derived proteins. A 75 kDa band is detected when assayed by immunoblot. This reagent cross reacts with human angiopoietin 1. Partially cross reactivity is noted with human angiopoietin 2.

Immunogen: This whole rabbit serum was prepared by repeated immunizations with a synthetic peptide N-Q-R-R-N-P-E-N-G-G-R-R-Y-N-R-I-Q-H-G-Q corresponding to a region (aa 21-40) near the N-terminus of mouse angiopoietin 1 protein conjugated to KLH using maleimide. A residue of cysteine was added to the amino terminal end to facilitate coupling.

General References:

Suri, C., Jones, P.F., Patan, S., Bartunkova, S., Maisonpierre, P.C., Davis, S., Sato, T.N. and Yancopoulos G.D. (1996) Requisite role of angiopoietin-1, a ligand for the TIE2 receptor, during embryonic angiogenesis. Cell, 87(7): 1171-1180.

Davis, S., Aldrich, T.H., Jones, P.F., Acheson, A., Compton, D.L., Jain, V., Ryan, T.E., Bruno, J., Radziejewski, C., Maisonpierre, P.C. and Yancopoulos, G.D. (1996) Isolation of angiopoietin-1, a ligand for the TIE2 receptor, by secretion-trap expression cloning. Cell 87:7, 1161-1169.

Maisonpierre, P.C., Suri, C., Jones, P.F., Bartunkova, S., Wiegand, S.J., Radziejewski, C., Compton, D., McClain, J., Aldrich, T.H., Papadopoulos, N., Daly, T.J., Davis, S., Sato, T.N. and Yancopoulos, G.D. (1997) Angiopoietin-2, a natural antagonist for Tie2 that disrupts in vivo angiogenesis. Science:277:5322, 55-60.

Note: For Research Use Only-not for use in or on humans or animals


Rabbit Anti-Mouse Angiopoietin 2 (Rabbit)   NEW

Cat# RDI-MANGIOP2abrx    $400.00/0.2ml

Presentation: 0.2ml neat rabbit sera in 0.02 M Potassium Phosphate, 0.15 M Sodium Chloride, pH 7.2 with 0.01% sodium azide

Lot #: see sheet with shipment

Protein Concen: 85 mg/ml (by Refractometry)

Storage Conditions: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening.

Storage Conditions: Store vial at -20° C prior to opening. Aliquot contents and freeze at -20° C or below for extended storage. Avoid cycles of freezing and thawing. Centrifuge product if not completely clear after standing at room temperature. This product is stable for several weeks at 4° C as an undiluted liquid. Dilute only prior to immediate use. Expiration date is one (1) year from date of opening.

Background Information: Angiopoeitin-1 (Ang-1) and Antiopoietin-2 (Ang2) are important for development of the endothelium, by regulating tyrosine phosphorylation of the membrane receptor Tie-2/Tek. Ang-1 binding to Tie-2/Tek causes phosphorylation of the receptor. Ang-2 competes for this binding, and thus blocks receptor phosphorylation. Ang-2 expression occurs at sites of vascular remodelling: dorsal aorta and major aortic branches, ovary, placenta and uterus.

Anti-Ang 1 and anti-Ang 2 antibodies recognize a band above 55 kDa. Both antibodies show banding at 75 kDa which resembles the original size, because these are soluble highly glycosylated proteins, which should run higher that the calculated molecular weight of 55 kD.

Application Note(s): Suitable for western blotting and other antibody based assays. A 1:500 dilution is recommended for western blotting. The reaction of this antiserum directly with cell supernatants may result in high background due to reactivity of components in the serum. This can be alleviated by first immunoprecipitating the antibody:antigen complex and then detecting the antigen. This method results in very clean and strong signal.

Recommended Dilutions: This product was assayed by immunoblot against supernatants of angiopoietin-expressing endothelial cells from human and mouse sources. A single band of the expected apparent molecular weight was observed at a 1:500 dilution. Researchers should determine optimal titers for other applications.

Purity and Specificity: This antiserum is directed against mouse angiopoietin 2 and shows no reactivity with angiopoietin 1 on mouse derived proteins. A 75 kDa band is detected when assayed by immunoblot. This reagent cross reacts with human angiopoietin 2. Partially cross reactivity is noted with human angiopoietin 1.

 

Immunogen: This whole rabbit serum was prepared by repeated immunizations with a synthetic peptide N-N-F-R-K-S-V-D-S-T-G-R-R-Q-Y-Q-V-Q-N-G-P corresponding to a region (aa 21-40) near the N-terminus of mouse angiopoietin 2 protein conjugated to KLH using maleimide. A residue of cysteine was added to the amino terminal end to facilitate coupling.

 

Related Product(s):

RDI-MANGIOP1abrX rabbit anti-angiopoietin 1 (mouse specific) $400.00/0.2ml

RDI-MANGIOP2abrx rabbit anti-Angiopoietin 2 (Mouse specific) (Rabbit) $400.00/0.2ml

RDI-MANGIOP1R-CP CONTROL PEPTIDE For Anti-ANGIOPOIETIN 1 (Mouse) $150.00

RDI-MANGIOP2R-CP CONTROL PEPTIDE For Anti-ANGIOPOIETIN 2 (Mouse) $150.00

 

General References:

Suri, C., Jones, P.F., Patan, S., Bartunkova, S., Maisonpierre, P.C., Davis, S., Sato, T.N. and Yancopoulos G.D. (1996) Requisite role of angiopoietin-1, a ligand for the TIE2 receptor, during embryonic angiogenesis. Cell, 87(7): 1171-1180.

Davis, S., Aldrich, T.H., Jones, P.F., Acheson, A., Compton, D.L., Jain, V., Ryan, T.E., Bruno, J., Radziejewski, C., Maisonpierre, P.C. and Yancopoulos, G.D. (1996) Isolation of angiopoietin-1, a ligand for the TIE2 receptor, by secretion-trap expression cloning. Cell 87:7, 1161-1169.

Maisonpierre, P.C., Suri, C., Jones, P.F., Bartunkova, S., Wiegand, S.J., Radziejewski, C., Compton, D., McClain, J., Aldrich, T.H., Papadopoulos, N., Daly, T.J., Davis, S., Sato, T.N. and Yancopoulos, G.D. (1997) Angiopoietin-2, a natural antagonist for Tie2 that disrupts in vivo angiogenesis. Science:277:5322, 55-60.

 

Note: For Research Use Only-not for use in or on humans or animals