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Mouse RANKL Recombinant Rabbit Monoclonal Antibody [PSH07-92] - BSA and Azide free (Detector)

Usd: 649

Specification

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Catalog# HA722924

Mouse RANKL Recombinant Rabbit Monoclonal Antibody [PSH07-92] - BSA and Azide free (Detector)

Application

  • ELISA(Det)

Reactivity

  • Mouse

Pair: Capture
Pair: Detector
Pair: Protein
Range

  • 8.2-2,000 pg/mL

PI

  • 6.46

Conjugation

  • unconjugated

Overview

Product Name

Mouse RANKL Recombinant Rabbit Monoclonal Antibody [PSH07-92] - BSA and Azide free (Detector)

Antibody Type

Recombinant Rabbit monoclonal Antibody

Immunogen

Recombinant protein within Mouse RANKL aa 158-316 (HA210558).

Species Reactivity

Mouse

Validated Applications

ELISA(Det)

Positive Control

Recombinant Mouse RANKL protein (HA210558).

Conjugation

unconjugated

Clone Number

PSH07-92

Product Features

Form

Liquid

Concentration

Storage Instructions

Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.

Storage Buffer

PBS (pH7.4).

Isotype

IgG

Purification Method

Protein A affinity purified.

Application Dilution

  • ELISA(Det)

  • Use at an assay dependent concentration. Can be paired for Sandwich ELISA with Rabbit monoclonal [PSH07-90] to Mouse RANKL antibody (Capture) (HA722922) or Rabbit monoclonal [PSH07-91] to Mouse RANKL antibody (Capture) (HA722923) and recombinant Mouse RANKL protein (HA210558) as the standard. The reference range value is 82.3-20,000 pg/ml (HA722922) or 8.2-9,000 pg/ml (HA722923).

Target

Function

Cytokine that binds to TNFRSF11B/OPG and to TNFRSF11A/RANK. Osteoclast differentiation and activation factor. Augments the ability of dendritic cells to stimulate naive T-cell proliferation. May be an important regulator of interactions between T-cells and dendritic cells and may play a role in the regulation of the T-cell-dependent immune response. May also play an important role in enhanced bone-resorption in humoral hypercalcemia of malignancy. Induces osteoclastogenesis by activating multiple signaling pathways in osteoclast precursor cells, chief among which is induction of long lasting oscillations in the intracellular concentration of Ca 2+ resulting in the activation of NFATC1, which translocates to the nucleus and induces osteoclast-specific gene transcription to allow differentiation of osteoclasts. During osteoclast differentiation, in a TMEM64 and ATP2A2-dependent manner induces activation of CREB1 and mitochondrial ROS generation necessary for proper osteoclast generation.

Background References

1. Mahoney D.J., Mikecz K., Ali T., Mabilleau G., Benayahu D., Plaas A., Milner C.M., Day A.J., Sabokbar A. TSG-6 regulates bone remodeling through inhibition of osteoblastogenesis and osteoclast activation. J. Biol. Chem. 283:25952-25962 (2008)

2. Decker C.E., Yang Z., Rimer R., Park-Min K.H., Macaubas C., Mellins E.D., Novack D.V., Faccio R. Tmem178 acts in a novel negative feedback loop targeting NFATc1 to regulate bone mass. Proc. Natl. Acad. Sci. U.S.A. 112:15654-15659 (2015)

Subcellular Location

Secreted.

UNIPROT

SwissProt: O35235 Mouse

Synonyms

CD254 antibody

hRANKL2 antibody

ODF antibody

OPGL antibody

OPTB2 antibody

Osteoclast differentiation factor antibody

Osteoprotegerin ligand antibody

Rank Ligand antibody

RANKL antibody

Receptor activator of nuclear factor kappa B ligand antibody

Expand

Images

  • Sandwich ELISA analysis of mouse RANKL matched pair antibodies<br /><br />Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (<a href="/products/HA722922" style="font-weight: bold;text-decoration: underline;">HA722922</a>) diluted in carbonate/bicarbonate buffer, at a concentration of 5 µg/ml overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1%BSA blocking buffer, and incubated with serial diluted Recombinant Mouse RANKL protein (<a href="/products/HA210558" style="font-weight: bold;text-decoration: underline;">HA210558</a>) starting from 8000 pg/ml to 0 pg/ml and detect antibody (<a href="/products/HA722924" style="font-weight: bold;text-decoration: underline;">HA722924</a>, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

    Sandwich ELISA analysis of mouse RANKL matched pair antibodies
    
    Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA722922) diluted in carbonate/bicarbonate buffer, at a concentration of 5 µg/ml overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1%BSA blocking buffer, and incubated with serial diluted Recombinant Mouse RANKL protein (HA210558) starting from 8000 pg/ml to 0 pg/ml and detect antibody (HA722924, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

  • Sandwich ELISA analysis of mouse RANKL matched pair antibodies<br /><br />Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (<a href="/products/HA722923" style="font-weight: bold;text-decoration: underline;">HA722923</a>) diluted in carbonate/bicarbonate buffer, at a concentration of 5 µg/ml overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1%BSA blocking buffer, and incubated with serial diluted Recombinant Mouse RANKL protein (<a href="/products/HA210558" style="font-weight: bold;text-decoration: underline;">HA210558</a>) starting from 8000 pg/ml to 0 pg/ml and detect antibody (<a href="/products/HA722924" style="font-weight: bold;text-decoration: underline;">HA722924</a>, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

    Sandwich ELISA analysis of mouse RANKL matched pair antibodies
    
    Elisa assay was performed by coating wells of a 96-well plate with 100 µl per well of capture antibody (HA722923) diluted in carbonate/bicarbonate buffer, at a concentration of 5 µg/ml overnight at 4℃. Wells of the plate were washed, blocked with 150 µl 0.05% tween-20 1%BSA blocking buffer, and incubated with serial diluted Recombinant Mouse RANKL protein (HA210558) starting from 8000 pg/ml to 0 pg/ml and detect antibody (HA722924, Biotin, 0.2 µg/ml) for 1 hour at 30℃ with shaking. Then the plate was washed and incubated with 100 µl per well of SA-HRP for 0.5 hour at 30℃ with shaking. Detection was performed using an Ultra TMB Substrate for 10 minutes at room temperature in the dark. The reaction was stopped with sulfuric acid and absorbances were read on a spectrophotometer at 450 nm.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Interpolated concentrations of native RANKL in mouse spleen cell culture supernatant treated or untreated with concanavalin A for 3days .<br /><br />Interpolated concentration of native RANKL was measured in duplicate at different sample concentrations and interpolated from the RANKL standard curves. Undiluted samples were 100% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean RANKL concentration was determined to be 3,251 pg/mL in neat mouse spleen cell treated supernatant, undetectable in untreated mouse spleen cell supernatant.

    ☑ Cell treatment (CT)

    Interpolated concentrations of native RANKL in mouse spleen cell culture supernatant treated or untreated with concanavalin A for 3days .
    
    Interpolated concentration of native RANKL was measured in duplicate at different sample concentrations and interpolated from the RANKL standard curves. Undiluted samples were 100% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean RANKL concentration was determined to be 3,251 pg/mL in neat mouse spleen cell treated supernatant, undetectable in untreated mouse spleen cell supernatant.

  • <span style="font-weight: bold;">☑ Cell treatment (CT)</span><br /><br />Interpolated concentrations of native RANKL in mouse spleen cell culture supernatant treated or untreated with concanavalin A for 3days .<br /><br />Interpolated concentration of native RANKL was measured in duplicate at different sample concentrations and interpolated from the RANKL standard curves. Undiluted samples were 100% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean RANKL concentration was determined to be 3,455 pg/mL in neat mouse spleen cell treated supernatant, undetectable in untreated mouse spleen cell supernatant.

    ☑ Cell treatment (CT)

    Interpolated concentrations of native RANKL in mouse spleen cell culture supernatant treated or untreated with concanavalin A for 3days .
    
    Interpolated concentration of native RANKL was measured in duplicate at different sample concentrations and interpolated from the RANKL standard curves. Undiluted samples were 100% cell supernatant. The interpolated dilution factor corrected values were plotted (mean +/- SD, n=2). The mean RANKL concentration was determined to be 3,455 pg/mL in neat mouse spleen cell treated supernatant, undetectable in untreated mouse spleen cell supernatant.

  • Interpolated concentrations of spiked RANKL in human cell culture media samples.<br /><br />The concentrations of RANKL were measured in duplicates, interpolated from the RANKL standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

    Interpolated concentrations of spiked RANKL in human cell culture media samples.
    
    The concentrations of RANKL were measured in duplicates, interpolated from the RANKL standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

  • Interpolated concentrations of spiked RANKL in cell culture media samples.<br /><br />The concentrations of RANKL were measured in duplicates, interpolated from the RANKL standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

    Interpolated concentrations of spiked RANKL in cell culture media samples.
    
    The concentrations of RANKL were measured in duplicates, interpolated from the RANKL standard curves and corrected for sample dilution. Undiluted samples are as follows: cell culture media 50%. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2).

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