Rabbit Anti Human SDF-1a polyclonal, antigen affinity

///Rabbit Anti Human SDF-1a polyclonal, antigen affinity

Rabbit Anti Human SDF-1a polyclonal, antigen affinity

$150.00$190.00

SKU: RKAP243 Category: Tags: , , , , , , , , , , ,

accession P48061


Applications

ELISA:This antibody can be used at 2 μg/mL jointly with biotinylated Human SDF-1a antibody to detect Human SDF-1a. The detection limit for recombinant Human SDF-1a 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 SDF-1a. The detection limit for recombinant Human SDF-1a is approximately 0.2 ng/lane.

Source This antibody was produced in rabbits immunized with recombinant Human SDF-1a.
Species reactivity Human
Purification The specific antibody was purified from Rabbit sera by using immobilized recombinant Human SDF-1a 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 P15172
Biological Process Chemotaxis
Molecular function Cytokine
Molecular function Growth-factor

Methods

CXCL12 ELISA

  • HBMEC or HUVECs (wild type or infected with viruses as described in results) were plated onto gelatin or Matrigel coated dishes and maintained in EGM-2MV media for 24 hours.
  • The media was then changed to Serum Free DMEM.
  • After 48 hours of additional incubation, culture supernatants were collected and concentrated (100×) for CXCL12 detection by indirect ELISA assay.
  • Briefly, CXCL12 standards, culture supernatants from endothelial cells and DMEM from Matrigel alone were incubated in a 96-well plate overnight in 20 mM sodium bicarbonate buffer (pH 9.5).
  • The plate was blocked with 2% BSA for 1 hour and then incubated with rabbit anti-hCXCL12 (1∶200 ) for 1 hour.
  • OPD substrate was added to the plate following incubation of goat anti-rabbit IgG HRP (1∶2500) for 1 hour.
  • The reaction was stopped by 3M HCL and OD at 490 nm was measured with a microplate reader .
  • CXCL12 concentrations were calculated with reference…
  • HBMEC or HUVECs (wild type or infected with viruses as described in results) were plated onto gelatin or Matrigel coated dishes and maintained in EGM-2MV media for 24 hours.
  • The media was then changed to Serum Free DMEM.
  • After 48 hours of additional incubation, culture supernatants were collected and concentrated (100×) for CXCL12 detection by indirect ELISA assay.
  • Briefly, CXCL12 standards, culture supernatants from endothelial cells and DMEM from Matrigel alone were incubated in a 96-well plate overnight in 20 mM sodium bicarbonate buffer (pH 9.5).
  • The plate was blocked with 2% BSA for 1 hour and then incubated with rabbit anti-hCXCL12 (1∶200 ) for 1 hour.
  • OPD substrate was added to the plate following incubation of goat anti-rabbit IgG HRP (1∶2500) for 1 hour.
  • The reaction was stopped by 3M HCL and OD at 490 nm was measured with a microplate reader .
  • CXCL12 concentrations were calculated with reference to an ELISA standard curve.
  • All samples were analyzed in duplicate.

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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