Rabbit Anti Human BMP2 polyclonal, antigen affinity

///Rabbit Anti Human BMP2 polyclonal, antigen affinity

Rabbit Anti Human BMP2 polyclonal, antigen affinity

$150.00$190.00

SKU: RKAP017 Category: Tags: , , , ,

accession P12643


Applications

ELISA:This antibody can be used at 2 μg/mL jointly with biotinylated Human Mouse Rat BMP2 antibody to detect Human Mouse Rat BMP2. The detection limit for recombinant Human Mouse Rat BMP2 is approximately 0.2 ng/well.

Western Blot:This antibody can be used at 0.2 μg/mL with the appropriate secondary reagents to detect Human Mouse Rat BMP2. The detection limit for recombinant Human Mouse Rat BMP2 is approximately 0.2 ng/lane.

Source This antibody was produced in Rabbit immunized with recombinant Human Mouse Rat BMP2.
Species reactivity Human Mouse Rat
Purification The specific antibody was purified from Rabbit sera by using immobilized recombinant Human Mouse Rat BMP2 affinity chromatography.
Presentation Lyophilized from PBS, pH 7.2.
Storage The lyophilized antibody is stable for at least 1 year from date of receipt at -20° C. Upon reconstitution, this antibody can be stored in working aliquots at 2° - 8° C for one month, or at -20° C for 12 months without detectable loss of activity. Avoid repeated freeze/thaw cycles.
Usage This antibody product is for research purposes only.It may not be used for therapeutics or diagnostic purposes.


Interactor O88273
Interactor P27037 AVR2A_HUMAN
Interactor Q13873 BMPR2_HUMAN
Interactor P27038 AVR2A_MOUSE
Interactor P36894 BMR1A_HUMAN
Interactor P37023 ACVL1_HUMAN
Interactor P97798
Biological Process Chondrogenesis
Biological Process Differentiation
Biological Process Osteogenesis
Molecular function Cytokine
Molecular function Developmental-protein
Molecular function Growth-factor

Methods

Western Blot

  • Ten µg of protein extract was loaded on acrylamide gel for SDS-PAGE electrophoresis.
  • Western blotting was carried out on nitrocellulose membrane blocked with PBS/0.05% Tween-20/2.5% BSA and probed with rabbit anti-BMP2 , mouse anti-BMP4 , goat anti-BMP5 , goat anti-BMP6 , rabbit anti-BMP7 overnight in PBS/0.05% Tween-20, 1% BSA.
  • Membranes were washed three times for 10 min each with PBS/0.05% Tween-20 and incubated with the appropriate secondary IgGs coupled to HRP.
  • Membranes were visualized using the ECL Plus detection kit on Biomax film .
  • To assess equal protein loading, membranes were stripped and re-probed with anti-GAPDH .

Production of recombinant growth factors in Nicotiana benthamiana

  • The sequences employed (TGFβ1 GenBank: X02812.1 TGFβ1, BMP2 NM_001200.2, BMP7 NM_001719.2) were optimized for expression in plants and synthesized by Genscript.com for cloning.
  • All sequences contained a 6xHis tag at the 3′end and were flanked with BsaI sites and cloned in the pICH31070 vector.
  • Orientation and codon in-frame for all constructs were confirmed by restriction analysis and sequencing.
  • All expression vectors were kindly provided by Dr. Yuri Gleba [

Sandwich ELISA

  • 96-well maxisorp plates were coated with the capture antibody overnight, then blocked with 5% FCS in PBS for 1 hour, then washed.
  • Then, the protein standard was added, and plates were incubated for 2 hrs at 37°C.
  • Plates were washed and incubated with a biotinylated detection antibody for 1 hour at 37°C.
  • After washing, plates were incubated with HRP-conjugated streptavidin for 30 min at room temperature and washed again.
  • Plates were developed using the tetramethylbenzidine peroxidase substrate system and absorbance was measured at 450-615 nm using an automated plate reader.

Western Blot

  • The Recombinant immunogen was migrated by 12% SDS-PAGE and transferred to a 0.2 um PVDF membrane.
  • After blocking the membrane in blocking buffer (5% milk powder in 20 mM Tris-HCl pH 7.5, 500 mM NaCl, 0.1% (v/v) Tween 20 , the membrane was incubated with primary antibody ( at 4°C overnight.
  • Peroxidase-linked secondary anti-rabbit were used to detect the bound primary antibodies.
  • Enhanced chemiluminescence (ECL) reagents were used to visualize