Recombinant human RANKL protein (Active)

• FuncS

Unknown

Functional Studies - Recombinant human RANKL protein (Active) (AB157289)

Binding of ab157289 to Osteoprotegerin. ab157289 binds to Osteoprotegerin : Fc. Method : Ligand binding assay : 96 well ELISA plates were coated O/N with 50 ng OPG-Fc per well. After a blocking step, the indicated concentrations of ab157289 were added for 1 hour. Bound ligand was revealed with an anti-FLAG antibody at 1 µg/ml for 30 minutes, followed by a rabbit anti-mouse IgG-HRP at 1/1000 dilution for 30 minutes. OPD was used as a substrate for the peroxidase. Absorbance was measured at 490 nm in an ELISA reader.

• FuncS

Unknown

Functional Studies - Recombinant human RANKL protein (Active) (AB157289)

Mononuclear cells differentiate into Osteoclasts in the presence of M-CSF and ab157289. Method : Human CD14+ mononuclear cells isolated from adult peripheral blood were cultured in control medium (upper panel), in 25ng/ml M-CSF (middle panel), or in 25ng/ml M-CSF plus 50ng/ml ab157289 (lower panel). Osteoclasts were identified as Tartrate-Resistant Acid Phosphatase (TRAP)-positive multinucleated cells. Osteoclasts were detected exclusively in presence of ab157289 and in these culture conditions (M-CSF + ab157289), cells fused and generated multinucleated dark red TRAP-positive cells (arrows). Nuclei were stained with haematoxylin.

• WB

Lab

Western blot - Recombinant human RANKL protein (Active) (AB157289)

All lanes:

Western blot - Anti-DDDDK tag (Binds to FLAG® tag sequence) antibody [6F7] (<a href='/en-us/products/primary-antibodies/ddddk-tag-binds-to-flag-tag-sequence-antibody-6f7-ab213519'>ab213519</a>) at 1 µg/mL

Lanes 1 and 3:

Western blot - Recombinant human RANKL protein (Active) (ab157289) at 0.5 µg with Milk in TBS-T

Lanes 2 and 4:

Western blot - Recombinant human RANKL protein (Active) (ab157289) at 0.1 µg with Milk in TBS-T

Lane 3:

Western blot - Recombinant human TRAIL protein (<a href='/en-us/products/proteins-peptides/recombinant-human-trail-protein-ab157345'>ab157345</a>) at 0.5 µg with Milk in TBS-T

Lane 4:

Western blot - Recombinant human TRAIL protein (<a href='/en-us/products/proteins-peptides/recombinant-human-trail-protein-ab157345'>ab157345</a>) at 0.1 µg with Milk in TBS-T

Secondary

All lanes:

Goat Anti-Rat IgG H&L (IRDye® 800CW ) (ab253031) at 1/20000 dilution

false

• SDS-PAGE

Unknown

SDS-PAGE - Recombinant human RANKL protein (Active) (AB157289)

SDS-PAGE analysis of ab157289. Lane 1 : MWt Marker, Lane 2 : ab157289 1µg.