AMBP | Bikunin (50 µl)

AS06 173 | Clonality: Polyclonal | Host: Rabbit | Reactivity: Human

Product Information

Immunogen

human recombinant bikunin, UniProt: P02760, cleavage product no. PRO_0000017887

Host Rabbit

Clonality Polyclonal

Purity Serum

Format Lyophilized

Quantity 50 µl

Reconstitution For reconstitution add 50 µl of sterile water

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

Tested applications ELISA (ELISA), Western blot (WB)

Recommended dilution 1 : 2000 (ELISA), 1 : 2000 (WB)

Expected | apparent MW

free bikunin 60-80 kDa

Reactivity

Confirmed reactivity Human

Predicted reactivity Bovine, Pig, Primates

Not reactive in No confirmed exceptions from predicted reactivity are currently known

Application examples

Application examples Application example

western blot using anti-human bikunin AMBP antibodies

Panel A: Coomasie stained gel. Panel B: 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) 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.

Additional information

Additional information This antibody is also reacting with the AMBP precursor

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 is found in blood and tissues both in free form and associated with heavy chains forming the inter-alpha-inhibitor family of proteins. Bikunin is a protease inhibitor, a component of extracellular matrix and exhibits anti-inflammatory activity in protection against cancer and inflammation.

Product citations

Selected references Olsson et al. (2007) Up-regulation of alpha1-microglobulin by hemoglobin and reactive oxygen species in hepatoma and blood cell lines.Free Radic Biol Med. 42(6):842-51.

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 is found in blood and tissues both in free form and associated with heavy chains forming the inter-alpha-inhibitor family of proteins. Bikunin is a protease inhibitor, a component of extracellular matrix and exhibits anti-inflammatory activity in protection against cancer and inflammation.
All references:	Olsson et al. (2007) Up-regulation of alpha1-microglobulin by hemoglobin and reactive oxygen species in hepatoma and blood cell lines.Free Radic Biol Med. 42(6):842-51.
calculated \| apparent molecular mass [kDa]:	free bikunin 60-80 kDa
Clonality:	Polyclonal
Format:	Lyophilized
Host:	Rabbit
immunogen:	human recombinant bikunin, UniProt: P02760, cleavage product no. PRO_0000017887
Purity:	Serum
Quantity:	50 µl
recommended dilution:	1 : 2000 (ELISA), 1 : 2000 (WB)
Reconstitution:	For reconstitution add 50 µl of sterile water
storage:	Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
tested applications:	ELISA (ELISA), Western blot (WB)
Picture (footer):	Application example Panel A: Coomasie stained gel. Panel B: 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) 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.
additional information:	This antibody is also reacting with the AMBP precursor
Confirmed reactivity:	Human
predicted reactivity:	Bovine, Pig, Primates
not reactive in:	No confirmed exceptions from predicted reactivity are currently known

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