Human GPNMB/Osteoactivin ELISA Kit PicoKine®

Osteoactivin/GPNMB ELISA kit for Human

Human GPNMB/Osteoactivin ELISA Kit PicoKine™ (96 Tests). Quantitate Human GPNMB in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml.

Product Info Summary

SKU: EK1251
Size: 96 wells/kit, with removable strips.
Reactive Species: Human
Application: ELISA
Sample Types: cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Product Name

Human GPNMB/Osteoactivin ELISA Kit PicoKine®

View all Osteoactivin/GPNMB ELISA kits

SKU/Catalog Number

EK1251

Size

96 wells/kit, with removable strips.

*Question: How many samples can I assay/run in this kit?

Description

Human GPNMB/Osteoactivin ELISA Kit PicoKine™ (96 Tests). Quantitate Human GPNMB in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml.

Storage & Handling

Store at 4°C for 6 months, at -20°C for 12 months. Avoid multiple freeze-thaw cycles (Ships with gel ice, can store for up to 3 days in room temperature. Freeze upon receiving.)

Cite This Product

Human GPNMB/Osteoactivin ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1251)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: NS0; Immunogen sequence: K23-P486

Sensitivity

<10 pg/ml

Assay Range

156 pg/ml - 10,000 pg/ml

Standard Dilution Instructions

serial dilution instructions image

Add 100ul of sample diluent in well #2-#8. Add 200ul standard stock solution to well #1, and serial dilute for well #2-#7 to make a standard curve row. Leave well #8 as blank See datasheet of EK1251 for more details

Cross-reactivity

There is no detectable cross-reactivity with other relevant proteins.

Reactive Species

EK1251 is reactive to GPNMB in Human samples

Validated Sample Types

cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Application Guarantee

EK1251 is guaranteed for ELISA in Human by Boster Guarantee

See how Boster Bio validate our ELISA kits: ELISA Validation Information

Background of Osteoactivin/GPNMB

Transmembrane glycoprotein NMB is a protein that in humans is encoded by the GPNMB gene. In osteoblast progenitor cells, GPNMB works as a positive regulator of osteoblast differentiation during later stages of matrix maturation and mineralization that is mediated at least in part by BMP-2 in a SMAD1 dependent manner to promote osteoblast differentiation. GPNMB can enhance the repairing process in bone fracture, demonstrated by its high expression during chondrogenesis (soft callus) and osteogenesis (hard callus) compared to the intact femurs that is why Osteoactivin (OA) could be a novel therapeutic agent used to treat generalized osteoporosis or localized osteopenia during fracture repair by stimulating bone growth and regeneration. Similarly, GPNMB expression increases during osteoclast differentiation and it is functionally implicated in this process, possibly by promoting the fusion of osteoclast progenitor cells.

Kit Components

Catalog Number Description Quantity
EK1251-CAP Anti-Human GPNMB Pre-coated 96-well strip microplate 1
EK1251-ST Human GPNMB Standard 2 vials, 10 ng/tube
EK1251-DA Human GPNMB Biotinylated antibody (100x) 100ul
AR1103 Avidin-Biotin-Peroxidase Complex (100x) 100ul
AR1106-1 Sample Diluent 30ml
AR1106-2 Antibody Diluent 12ml
AR1106-3 Avidin-Biotin-Peroxidase Diluent 12ml
AR1104 Color Developing Reagent (TMB) 10ml
AR1105 Stop Solution 10ml
AR1106-5 Wash Buffer (25x) 20ml
PLA-SEA Adhesive plate sealers 4

*The kit components are not available for individual purchase.

Materials Required But Not Included In Kit

  • Microplate Reader capable of reading absorbance at 450nm.
  • Incubator.
  • Automated plate washer (optional).
  • Pipettes and pipette tips capable of precisely dispensing 0.5 µl through 1 ml volumes of aqueous solutions.
  • Multichannel pipettes are recommended for large amount of samples.
  • Deionized or distilled water.
  • 500ml graduated cylinders.
  • Test tubes for dilution.

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Validation Standard Curve O.D. At 450nm

Concentration (pg/ml)015631262512502500500010000
O.D.0.0280.0560.1060.1700.3110.5421.1691.880

Data Example Images

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Human GPNMB/Osteoactivin ELISA Kit PicoKine® within the same batch/lot to ensure the consistency of the kits' performances. ELISA intra assay consistency is measured using wells from the same plate/assay kit. ELISA inter assay consistency is measured using wells from different plates from the same batch production/lot.

Intra-Assay PrecisionInter-Assay Precision
Sample123123
n161616242424
Mean (pg/ml)2612107561226022875804
Standard deviation12.26124.31432.1212.48169.23539.77
CV (%)4.7%7.7%7.7%4.8%7.4%9.3%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of Osteoactivin/GPNMB in ELISA kits from four different production batches/lots.

LotsLot 1 (pg/ml)Lot 2 (pg/ml)Lot 3 (pg/ml)Lot 4 (pg/ml)Mean (pg/ml)Standard DeviationCV (%)
Sample 126122322524523815.586.5%
Sample 221071918179717741899131.966.9%
Sample 356124864583155475463361.756.6%
*number of samples for each test n=16.

Gene/Protein Information For GPNMB (Source: Uniprot.Org, NCBI)

Gene Name

GPNMB

Full Name

Transmembrane glycoprotein NMB

Weight

63.923kDa

Superfamily

PMEL/NMB family

Alternative Names

DC-HIL; glycoprotein (transmembrane) nmb; glycoprotein NMB; GPNMB; HGFIN; HGFINglycoprotein nmb-like protein; NMBosteoactivin; Osteoactivin; Transmembrane glycoprotein HGFIN; transmembrane glycoprotein NMB GPNMB HGFIN, NMB, PLCA3 glycoprotein nmb transmembrane glycoprotein NMB|glycoprotein (transmembrane) nmb|glycoprotein nmb-like protein|glycoprotein nonmetastatic melanoma protein B|hematopoietic growth factor inducible neurokinin-1 type|osteoactivin|transmembrane glycoprotein HGFIN

*if product is indicated to react with multiple species, protein info is based on the gene entry specified above in "species".

For more info on GPNMB, check out the GPNMB Infographic

GPNMB infographic

We have 30,000+ of these available, one for each gene! check them out.

In this infographic you will see the following information for GPNMB: database IDs, super-family, protein function, synonyms, molecular weight, chromosomal locations, tissues of expression, subcellular locations, post translational modifications, and related diseases, research areas & pathways. If you want to see more information included, or would like to contribute to it and be acknowledged, please contact us [email protected].

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7 Customer Q&As for Human GPNMB/Osteoactivin ELISA Kit PicoKine®

Question

Q: can you recommend the dilution ratio of serum samples for detection of Osteoactivin in Human cell lysates? I am trying to measure a a number of analytes and it requires 100ul of diluted samples for each well. We have limited sample quantitys so we like to dilute as much as possible.

Verified Customer

Verified customer

Asked: 2020-07-20

Answer

A: unable to know the physiological or pathological context of your samples we cannot recommend a dilution ratio without performing a pilot test with your samples. Here is how you can perform a pilot study on your own: perform a serial dilution of your samples on the Osteoactivin ELISA kit to make sure you have a linear ascending curve followed by a plateau, which signifies the samples saturating the detection limit of the kit. Then you can pick the dilution ratios from samples in the linear part of the curve as your experimental dilution ratio.
If you are interested in using our ELISA service, you can also send us your sample and we will take care of everything for you. You can check our service details here: bosterbio.com/services/assay-services/ELISA-testing-service
Since you mentioned you have limited samples, our cost effective multiplex ELISA service would fit perfectly for your needs, where we can generate dozens of data points using as little as 25ul sample volume. Information on this service is also in the above link.

Boster Scientific Support

Answered: 2020-07-20

Question

Q: What is the optimal O.D. value for Osteoactivin ELISA kit? I used your Osteoactivin ELISA on serum samples. For my positive control, I received an O.D. value of 0.826, while my negative control received a value of 0.136. I obtained both of these controls from the ELISA kit, where your kit's typical data shows O.D. values much higher than my positive control and your background is lower. My samples O.D. values are around 0.225 and the highest is only 0.357. can I consider these samples contain Osteoactivin even though the O.D. values are not very high?

Verified Customer

Verified customer

Asked: 2020-06-25

Answer

A: The absolute O.D. values may change according to incubation time. The more you incubate the higher the O.D. values are going to be. a point of focus should be is whether your sample O.D. values are statistically significantly higher than your blank values. in the example mentioned, you could extend your development time in the substrate incubation step to obtain higher O.D. values, as long as your negative controls' O.D. values are not increasing faster in relation to your positive controls. normally, a sample with O.D. value 2 standard deviations higher than your negative controls can be considered positive. We calculate the sensitivity of this ELISA kit by changeing cutoff O.D. value, calculated as the average of 20 negative controls plus 2 standard deviations of the 20 negative controls, into a concentration. in other words, when we claim this Osteoactivin ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of Osteoactivin that can be declared/interpreted as positive by the above standard is 10pg/ml.

Boster Scientific Support

Answered: 2020-06-25

Question

Q: Are Boster Bio recombinant proteins and antibodies sterile?

W. Hall

Verified customer

Asked: 2020-04-21

Answer

A: although the vials are bottled using aseptic techniques, heat-treated vials, and sterile stock solutions, they are not considered or guaranteed to be sterile. If sterile material is needed for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.

Boster Scientific Support

Answered: 2020-04-21

Question

Q: is there any online tool I can use to streamline the data analysis for my ELISA results?

Verified Customer

Verified customer

Asked: 2018-12-19

Answer

A: We have a web based ELISA curve fitting (4pl) and data analysis tool. Please give it a try: bosterbio.com/biology-research-tools/ELISA-data-analysis-online. You can also consult our article on ELISA data analysis: bosterbio.com/ELISA-data-analysis-instructions

Boster Scientific Support

Answered: 2018-12-19

Question

Q: how much samples can be assayed in a Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2018-06-25

Answer

A: The Picokine® ELISA Kits will generally run a 7-point standard curve, non-specific binding wells, and 39 samples in duplicate. this may vary slightly by kit so please refer to each datasheet for details.

Boster Scientific Support

Answered: 2018-06-25

Question

Q: if the enzyme conjugated Osteoactivin antibodies are mixed with the substrate, will that change the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the Osteoactivin antigen?

P. Lee

Verified customer

Asked: 2018-04-11

Answer

A: The enzyme is always active. Avoid contaminating the substrate with enzyme prior to the incubation otherwise it compromises the assay with false positive signal.

Boster Scientific Support

Answered: 2018-04-11

Question

Q: Can Osteoactivin ELISA Kits be used with tissue homogenates (or other non-validated sample types)?

Verified Customer

Verified customer

Asked: 2014-04-24

Answer

A: Unfortunately, Boster Bio has not routinely validated tissue homogenates as a sample type for ELISA kits. This does not mean that ELISA kits are not valid for other sample types than we have tested: it means further investigation is needed. One will need to perform a spike and recovery study to determine if an unvalidated sample type will work with a particular kit. To perform a spike and recovery experiment, one should divide a sample into two aliquots. In one of the aliquots, the user should spike in a known amount of the kit standard. a dilution series is performed comparing the spiked versus the unspiked sample. Generally, samples with expected recovery and linearity between 80-120% are considered acceptable. This method can be used to validate any sample type that has not been assessd by Boster Bio. for a more detailed spike and recovery protocol, please contact technical support.
Note: acceptable ranges should be determined individually by each laboratory. Additionally, technical support can help determine if a buffer component is not compatible with a given ELISA kit. please check the Citations tab on the product webpage for peer-reviewed papers utilizing a wide range of sample types. We also have an innovator's reward program where if the user validates our ELISA kits in applications or samples previously not validated by Boster Bio or other users, and share such information with us by submit a review, we will reward the user's efforts with a free antibody or ELISA kit from our catalog. Biocompare.com will also give $20 Amazon giftcard as an additional reward, if the review is submitted there as well.

Boster Scientific Support

Answered: 2014-04-24

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