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AMBP | Bikunin

405 €

AS04 039  |  clonality: polyclonal  |  host: rabbit  |  reactivity: rat

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Item No:
AS04 039

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product information
Background

Bikunin is a Kunitz-type protease inhibitor synthesized in the liver. The AMBP gene resides in the lipocalin cluster and encodes alpha-1-microglobulin together with a Kunitz-type proteinase inhibitor, bikunin. The gene is translated into the alpha-1-microglobulin-bikunin precursor, which is subsequently cleaved and the two proteins are secreted to the blood separately. Bikunin exhibits anti-inflammatory activity in protection against cancer and inflammation.

Immunogen

Recombinant rat bikunin purified from baculovirus infected insect cell cultures

Host Rabbit
Clonality Polyclonal
Clone
Purity Serum
Format Lyophilized
Quantity 200 ĩl
Reconstitution For reconstitution add 200 ĩl of sterile water
Storage

store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

Tested applications Western blot (WB), ELISA (ELISA), immunohistochemistry (IHC)
Related products

AS04 040 | anti-mice bikunin

AS06 173 | anti-human bikunin

Secondary antibodies

Additional information

This antibody is also reacting with the AMBP precursor.

application information
Recommended dilution

1: 2000 (WB), 1: 2000 (ELISA ), use at an assay dependent dilution (IHC)

Expected | apparent MW

Free bikunin appears as a smear between 60-80 kDa, because the protein core carries sulfated proteoglycans.

The most common bikunin complexes are called pre-alpha-inhibitor (PaI) and inter-alpha-inhibitor (IaI) and appears as approximately 120 and 180 kDa

Confirmed reactivity

rat

Predicted reactivity

mouse

Not reactive in

no confirmed exceptions from predicted reactivity known in the moment

Additional information

to be added when available

Selected references

to be added when available


Application example

western blot using anti/rat bikunin antibodies

5 ug human (1), rat (2) or mouse (3) bikunin, expressed and purified from recombinant insect cells as described (ref 1), 8 µg of total protein from human (4), rat (5) or mouse (6) serum were mixed in sample buffer, containing 2% SDS and 1% mercaptoethanol, and denatured at 100°C for 2 min, and then separated on pre-cast 4-20 % gels bought from Bio-Rad (Mini-PROTEAN Tris-Glycine, Stain-Free, 4-20%) and blotted PVDF using semi-dry transfer. The membranes were blocked with 1% gelatin buffer 20 min at room temperature (RT) with agitation, and then incubated in the primary antibody at a dilution of 1: 2000 in 0.5% gelatin and 0.25% Tween-20 overnight at 4°C with agitation. The antibody solution was decanted and the membrane was rinsed 4x7 min with in 0.5% gelatin and 0.25% Tween-20, and incubated in secondary antibody (goat anti-rabbit IgG-Alexa 647, from Molecular Probes Inc) diluted to 10 ug/ml in 0.5% gelatin and 0.25% Tween-20 for 1h at RT with agitation. The blot was washed as above and developed using a ChemiDoc™ MP System (Bio-Rad) with the Alexa 650-settings.

Free bikunin appears as a smear between 60-80 kDa, because the protein core carries sulfated proteoglycans.
The most common bikunin complexes are called pre-alpha-inhibitor (PaI) and inter-alpha-inhibitor (IaI) and appears as  approximately 120 and 180 kDa, respectively. Zho et al. (2004). Inter-alpha-trypsin inhibitor, a covalent protein-glycosaminoglycan-protein complex. J Biol Chem. 2004 Sep 10;279(37):38079-82.

Reference 1. Falkenberg et al. (2001). Expression of a functional proteinase inhibitor capable of accepting xylose: bikunin. Arch Biochem Biophys 387: 99-106

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