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Mouse Osteoactivin/GPNMB DuoSet ELISA

Catalog #: DY2330
11 Citations Datasheet / COA / SDS
Catalog # Availability Size / Price Qty
DY2330
Ancillary Products Available
DY008B: DuoSet ELISA Ancillary Reagent Kit 2
DY999B: Substrate Reagent Pack
DY999: Substrate Reagent Pack
Mouse Osteoactivin / GPNMB ELISA Standard Curve
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Citations (11)
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Summary Product Features Kit Content Other Reagents Required Data Examples Product Datasheets Preparation and Storage Background

Mouse Osteoactivin/GPNMB DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Sample Volume Required
100 µL
Assay Range
0.3 - 20 ng/mL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant mouse Osteoactivin/GPNMB. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet.

 

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Scientific Data

N/A Mouse Osteoactivin / GPNMB ELISA Standard Curve View Larger

Mouse Osteoactivin / GPNMB ELISA Standard Curve

Product Datasheets

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Product Datasheet
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SDS

Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Osteoactivin/GPNMB

Osteoactivin, also known as transmembrane glycoprotein NMB (GPNMB) and dendritic cell-associated transmembrane protein (DC-HIL), is a type I transmembrane glycoprotein that is found either on the plasma membrane or on membranes of the endosome/lysosome compartment. The extracellular region of Osteoactivin contains a heparin-binding motif and an integrin recognition RGD domain, while the cytocylasmic region contains an immunoreceptor tyrosine-based activation motif (ITAM) and lysosomal targeting motif.

Long Name:
Glycoprotein Non-Metastatic Melanoma Protein B
Entrez Gene IDs:
10457 (Human); 93695 (Mouse); 113955 (Rat); 102133517 (Cynomolgus Monkey)
Alternate Names:
DC-HIL; glycoprotein (transmembrane) nmb; glycoprotein NMB; GPNMB; HGFIN; HGFINglycoprotein nmb-like protein; NMBosteoactivin; Osteoactivin; Transmembrane glycoprotein HGFIN; transmembrane glycoprotein NMB

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

 

Citations for Mouse Osteoactivin/GPNMB DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

11 Citations: Showing 1 - 10
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  1. ASO-mediated knockdown of GPNMB in mutant- GRN and Grn -deficient peripheral myeloid cells disrupts lysosomal function and immune responses
    Authors: Wallings, RL;Gillett, DA;Staley, HA;Mahn, S;Mark, J;Neighbarger, N;Kordasiewicz, H;Hirst, WD;Tansey, MG;
    bioRxiv : the preprint server for biology
    Species: Mouse
    Sample Types: Cell Culture Supernates
  2. Intracerebroventricular Treatment with 2-Hydroxypropyl-&beta-Cyclodextrin Decreased Cerebellar and Hepatic Glycoprotein Nonmetastatic Melanoma Protein B (GPNMB) Expression in Niemann-Pick Disease Type C Model Mice
    Authors: M Fukaura, Y Ishitsuka, S Shirakawa, N Ushihama, Y Yamada, Y Kondo, T Takeo, N Nakagata, K Motoyama, T Higashi, H Arima, Y Kurauchi, T Seki, H Katsuki, K Higaki, M Matsuo, T Irie
    International Journal of Molecular Sciences, 2021-01-05;22(1):.
    Species: Mouse
    Sample Types: Serum
  3. Glycoprotein NMB: a novel Alzheimer's disease associated marker expressed in a subset of activated microglia
    Authors: M Hüttenrauc, I Ogorek, H Klafki, M Otto, C Stadelmann, S Weggen, J Wiltfang, O Wirths
    Acta Neuropathol Commun, 2018-10-19;6(1):108.
    Species: Mouse
    Sample Types: Tissue Homogenates
  4. HEPES activates a MiT/TFE-dependent lysosomal-autophagic gene network in cultured cells: A call for caution
    Authors: MJ Tol, MJC van der Li, TL Gabriel, JJ Hagen, S Scheij, T Veenendaal, J Klumperman, WE Donker-Koo, AJ Verhoeven, H Overkleeft, JM Aerts, CA Argmann, M van Eijk
    Autophagy, 2018-02-17;0(0):1-13.
    Species: Mouse
    Sample Types: Cell Culture Supernates
  5. Validating glycoprotein non-metastatic melanoma B (gpNMB, osteoactivin), a new biomarker of Gaucher disease
    Authors: Vagishwari Murugesan
    Blood Cells Mol. Dis, 2016-12-13;0(0):.
    Species: Mouse
    Sample Types: Serum
  6. Elevation of glycoprotein nonmetastatic melanoma protein B in type 1 Gaucher disease patients and mouse models
    FEBS Open Bio, 2016-07-30;6(9):902-13.
    Species: Mouse
    Sample Types: Plasma
  7. Inhibition of Phosphatase Activity Follows Decline in Sulfatase Activity and Leads to Transcriptional Effects through Sustained Phosphorylation of Transcription Factor MITF
    Authors: S Bhattachar, L Feferman, JK Tobacman
    PLoS ONE, 2016-04-14;11(4):e0153463.
    Species: Mouse
    Sample Types: Cell Lysates
  8. Gpnmb Is a Potential Marker for the Visceral Pathology in Niemann-Pick Type C Disease.
    Authors: Marques A, Gabriel T, Aten J, van Roomen C, Ottenhoff R, Claessen N, Alfonso P, Irun P, Giraldo P, Aerts J, van Eijk M
    PLoS ONE, 2016-01-15;11(1):e0147208.
    Species: Mouse
    Sample Types: Plasma
  9. Identification of a biomarker in cerebrospinal fluid for neuronopathic forms of Gaucher disease.
    Authors: Zigdon, Hila, Savidor, Alon, Levin, Yishai, Meshcheriakova, Anna, Schiffmann, Raphael, Futerman, Anthony
    PLoS ONE, 2015-03-16;10(3):e0120194.
    Species: Mouse
    Sample Types: CSF
  10. Lysosomal stress in obese adipose tissue macrophages contributes to MITF-dependent Gpnmb induction.
    Authors: Gabriel T, Tol M, Ottenhof R, van Roomen C, Aten J, Claessen N, Hooibrink B, de Weijer B, Serlie M, Argmann C, van Elsenburg L, Aerts J, van Eijk M
    Diabetes, 2014-05-01;63(10):3310-23.
    Species: Mouse
    Sample Types: Plasma
  11. Mutation in osteoactivin decreases bone formation in vivo and osteoblast differentiation in vitro.
    Authors: Abdelmagid S, Belcher J, Moussa F, Lababidi S, Sondag G, Novak K, Sanyurah A, Frara N, Razmpour R, Del Carpio-Cano F, Safadi F
    Am J Pathol, 2014-01-23;184(3):697-713.
    Species: Mouse
    Sample Types: Serum

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