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Western-Ligand Blotting Kit for Human<br />(Pig, Cow, Goat, Sheep, Guinea Pig, Rabbit, Donkey) IGFBP´s
Western-Ligand Blotting Kit for Human and other species IGFBP´s
Reacts with Pig, Cow, Goat, Sheep, Guinea Pig, Rabbit, Donkey IGFBP´s
NON-RADIOACTIVE WESTERN-LIGAND BLOTTING KITS FOR THE DETECTION OF IGFBPs
SOME EXAMPLES OF WESTERN-LIGAND BLOTS WITH HUMAN IGF-I, IGF-II AND ANALOGS CAN BE SEEN IN APPLICATION NOTE IGF004:
A COMPARISON OF WESTERN LIGAND BLOT AND IMMUNOBLOT WITH IGFBP-3 AS AN EXAMPLE CAN BE SEEN IN
APPLICATION NOTE IGF0018:
Human western-ligand blotting kit
Detects IGFBP's and IGF-Receptors in samples of human origin. Similar patterns are obtained with samples from bovine, porcine, rabbit, guinea pig, sheep, donkey and goat serum. Only weak bands are obtained with rat and mouse serum on nitrocellulose membranes (see: Mouse/rat western-ligand blotting kit).
The kit is suitable for quatitative analysis. This is offered as a service by Ligandis GbR. Link to Ligandis.
Contents of the kit:
The reagents of the kit are sufficient for the preparation of 250 ml of each reagent in each step. This is sufficient for 500 square-cm of blotting membrane. Each reagent of the kit is also available separately.
| Reagent | Description | Quantity | Product Code | Price € |
| A | 0.5 ml Biotinylated ligand: biotinylated human IGF-II, dilute 1 : 100 to 1 : 500 | 10 ug | BioIGF2-10 | 150.- |
| B | Quenching Buffer, 2-fold conc. | 125 ml | QB-125 | 15.50 |
| C | Blocking/Dilution Buffer, 10 - fold conc. | 75 ml | BDB-75 | 50.- |
| D | Washing Buffer, 20-fold conc. | 125 ml | Product Code: WB-125 | 35.- |
| E | Streptavidin-POD-Conjugate, dilute 1:2500 - 1:5000 | 125 µl | Product Code: SPCON | 67.- |
| Detailed working instructions | Application Note IGF005 | |||
| Human Western-ligand Blot | 1 Kit | Product Code: human-WLB | 260.- |
Our kits are supplied without substrates, you can use your usual substrate
We have tested other substrates successfully, as e.g. KPL, Pierce, GE healthcare, Seramun, Kem-en-tec.
A recipe for a home-made chemiluminescent substrate is available (in German Language) at:
http://www.laborjournal.de/rubric/tricks/tricks/trick81.html
The colorimetric substrate can be used after the chemiluminescent substrate to give a permanent record. The membrane is simply washed for 1 min and the incubated with the colorimetric substrate. The sensitivity of the colorimetric substrate is similar to the chemiluminescent substrate, but incubation times are longer.
NOTES ON USE OF YOUR OWN BUFFERS
We have developped a buffer system, giving an optimal sensitivity on nitrocellulose membranes in our hands and having a prolonged stability, even at room temperature. Other buffers may propably be used, with a few limitations:
- Take care of the quality of bovine serum albumin (BSA), if you use it for blocking and/or dilution. Many BSA preparations contain traces of IGFBPs, that may affect the quality (Background/signal ratio) of the WLB's.
- In literature it is described that milk contains biotinylated proteins, that might affect the quality (background/signal ratio) of the WLB's. We have tested non-fat dry milk from different suppliers and from our local supermarket and found no biotinylated proteins.
Reference:
Pircher et al.: A new method for the purification of bioactive insulin-like growth factor-binding protein-3. Protein Expression and Purification 71 (2010) 160–167.
Link to Abstract:
http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6WPJ-4YC80SN-2&_user=10&_coverDate=06%2F30%2F2010&_rdoc=1&_fmt=high&_orig=search&_sort=d&_docanchor=&view=c&_searchStrId=1428943115&_rerunOrigin=google&_acct=C000050221&_version=1&_urlVersion=0&_userid=10&md5=f7f1e043cbca904e86719b7143f20bf9
Rev. 2013-02
Datasheet for Western-Ligand Blotting Kit for Human and other species IGFBP´s