Mouse Anti-Human PlGF-2
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| Cat-Nr. | 101-M65A |
| Size | 100 µg |
| Price | 275 € |
| Category | Monoclonal Antibody |
| Clone Nr. | (#A6D1) |
| Isotype | IgG1 |
| Species Reactivity | Human |
| Formulation | lyophilized |
| Buffer | PBS |
| Reconstitution | Centrifuge vial prior to opening. Reconstitute in sterile water to a concentration of 0.1-1.0 mg/ml. |
| Stability and Storage | The lyophilized antibody is stable for at least 2 years at -20°C. After sterile reconstitution the antibody is stable at 2-8°C for up to 6 months. Frozen aliquots are stable for at least 6 months when stored at -20°C. Addition of a carrier protein or 50% glycerol is recommended for frozen aliquots. |
| Preparation | The monoclonal antibody was produced with the help of BALB/c mice using recombinant human PlGF-2 (produced in insect cells) as the immunizing antigen. Mouse IgG1 antibody (clone #3B10) from hybridomas was purified from cell culture supernatant by Protein G chromatography. |
| Antigen | Recombinant human PlGF-2 (RT #300-019) |
| Application | E, WB, IF, IHC |
| Synonyms | PlGF; placental growth factor |
| Description | Placenta Growth Factor-2 (PlGF-2), a 22 kDa protein consisting of 152 amino acid residues is produced as a homodimer. PlGF is a polypeptide growth factor and a member of the platelet-derived growth factor family but more related to vascular endothelial growth factor (VEGF). PlGF acts only as a weak mitogen for those cell types possessing receptors for binding (e.g. vascular endothelial cells). At least one high-affinity receptor for PlGF (FLT-1 or VEGF-R1) has been demonstrated in different primary cell types (e.g. human umbilical vein endothelial cells and monocytes). In addition to its action as a weak mitogen it is also a chemoattractant for monocytes and endothelial cells. Three different proteins are generated by differential splicing of the human PlGF gene: PlGF-1 (131aa native chain), PlGF-2 (152aa) and PlGF-3 (203aa). All 3 mitogens are secretable proteins, but PlGF-2 can bind to heparin with high affinity. PlGF is apparently a homodimer, but preparations of PlGF show some heterogeneity on SDS gels depending of the varying degrees of glycosylation. All dimeric forms possess similar biological activities. Heterodimers between VEGF and PlGF are mitogenic for endothelial cells and have strong angiogenic activity in vivo (e.g. in the CAM assay or in the cornea pocket assay). Different cells and tissues (e.g. placenta) express PlGF-1 and PlGF-2 at different rates. A much related protein of PlGF is VEGF with about 53% homology and VEGF-B with similar biological activities. |
| Uniprot ID | P49763 |
| Protein RefSeq | NP_001193941.1 |
| mRNA RefSeq | NM_001207012.1 |
Figures
Western Blot analysis with human PlGF1 and PlGF2. Purified recombinant human PlGF1 (red: 500ng, non-red: 1000ng) and PlGF2 (red: 250ng, non-red: 500ng) were loaded in 15% SDS-PAGE under reducing and non-reducing conditions. As primary antibody mouse anti-human PlGF2 #D1 [Cat# 101-M65A] was used in a concentration of 0.5µg/ml. The detection was carried out with a secondary, AP-conjugated anti-mouse Fc antibody (Dianova, 1:1000). NOTE: The anti-human PlGF-2 #D1 antibody [Cat# 101-M65A] recognizes solely the PlGF2 isoform.
Western Blot analysis with PlGF. Purified recombinant human PlGF2 (100 ng/lane) derived from insect cells was loaded in 12.5% SDS-PAGE under reducing conditions. As primary antibody mouse anti-human PlGF2 #D1 [Cat# 101-M65A] was used in a concentration of 1µg/ml (A), 0.5µg/ml (B), 0.1µg/ml (C) and 0.05µg/ml (D). The detection was carried out with a secondary, AP-conjugated anti-mouse Fc antibody. NOTE: There is still a signal visible for PlGF2 at an antibody concentration of 50ng/ml.
ELISA with recombinant human PlGF1 and PlGF2 derived from insect cells. Both proteins were coated to a 96-well plate in a concentration of 0.2µg/ml. The mouse anti-human PlGF #331/H12 antibody [Cat# 101-M69] recognizing both isoforms and the mouse anti-human PlGF2 #D1 antibody [Cat# 101-M65A] recognizing specifically PlGF2 were added with increasing concentrations. Detection was carried out with an anti-mouse Biotin- conjugated antibody (Dianova).
Western Blot analysis with PlGF. Purified recombinant human PlGF2 (500 -10 ng/lane), PlGF1 (1000 ng/lane), and mouse and rat PlGF (500ng/lane) all derived from insect cells were loaded in 12.5% SDS-PAGE under reducing conditions. As primary antibody mouse anti-human PlGF2 #D1 [Cat# 101-M65A] was used in a concentration of 2µg/ml. The detection was carried out with a secondary, AP-conjugated anti-mouse Fc antibody. NOTE: There is no cross reaction with human PlGF1, mouse and rat PlGF.
IF staining of human PlGF1 and PlGF2 in Sf9 cells expressing either PlGF1 or PlGF2. Upper lane: mouse anti-human PlGF2 #D1 [Cat# 101-M65A] recognizing only PlGF2 and lower lane: mouse anti-human PlGF #178/G10 [Cat# 101-M67] recognizing both isoforms of PlGF.
ELISA with recombinant human PlGF1 and PlGF2 derived from insect cells. Both proteins were coated to a 96-well plate in increasing amounts. The mouse anti-human PlGF #178/G10 antibody [Cat# 101-M67] recognizing both isoforms and the mouse anti-human PlGF2 #D1 antibody [Cat# 101-M65A] recognizing specifically PlGF2 were added with a concentration of 0.5µg/ml. Detection was carried out with an anti-mouse Biotin antibody (Dianova).
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