Rabbit Anti Human SCF polyclonal, antigen affinity

///Rabbit Anti Human SCF polyclonal, antigen affinity

Rabbit Anti Human SCF polyclonal, antigen affinity

$150.00$190.00


accession P21583


Applications

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

Source This antibody was produced in rabbits immunized with recombinant Human SCF.
Species reactivity Human
Purification The specific antibody was purified from Rabbit sera by using immobilized recombinant Human SCF 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.


Biological Process Cell-adhesion
Molecular function Growth-factor

Methods

Treatment schema

  • When the tumors were about 5 mm in length (after about 6 days of 4T1 injections), mice were injected (IP) with neutralizing rat anti-mouse cKit-receptor antibody (50 μg) (eBioscience) or 20 μg goat anti-mouse-SCF antibody once a week for 4 weeks.
  • The mice were euthanized 24 hours after the last injections at about 35 days after tumor challenge.
  • For treatment experiments, 4T1 cells transfected with green fluorescent protein (4T1-GFP) were injected.

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