Rat ADAM12 ELISA Kit PicoKine®

ADAM12 ELISA kit for Rat

Rat ADAM12 ELISA Kit PicoKine™ (96 Tests). Quantitate Rat Adam12 in cell culture supernatants, serum and plasma (heparin). Sensitivity: 10pg/ml.

Product Info Summary

SKU: EK1293
Size: 96 wells/kit, with removable strips.
Reactive Species: Rat
Application: ELISA
Sample Types: cell culture supernatants, serum and plasma (heparin).

Product Name

Rat ADAM12 ELISA Kit PicoKine®

View all ADAM12 ELISA kits

SKU/Catalog Number

EK1293

Size

96 wells/kit, with removable strips.

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

Description

Rat ADAM12 ELISA Kit PicoKine™ (96 Tests). Quantitate Rat Adam12 in cell culture supernatants, serum and plasma (heparin). 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

Rat ADAM12 ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1293)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: CHO; Immunogen sequence: R29-S513

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 EK1293 for more details

Cross-reactivity

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

Reactive Species

EK1293 is reactive to ADAM12 in Rat samples

Validated Sample Types

cell culture supernatants, serum and plasma (heparin).

Application Guarantee

EK1293 is guaranteed for ELISA in Rat by Boster Guarantee

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

Background of ADAM12

ADAM12 (Disintegrin and metalloproteinase domain-containing protein 12), also known as MLTN, is an enzyme that in humans is encoded by the ADAM12 gene. This gene encodes a member of the ADAM (a disintegrin and metalloprotease) protein family. Members of this family are membrane-anchored proteins structurally related to snake venom disintegrins, and have been implicated in a variety of biological processes involving cell-cell and cell-matrix interactions, including fertilization, muscle development, and neurogenesis. This gene has two alternatively spliced transcripts: a shorter secreted form and a longer membrane-bound form. The shorter form is found to stimulate myogenesis. By RT-PCR and immunoblot analyses that expression of mouse Adam12 increases during muscle regeneration, while the levels of other ADAMs remain constant. Immunofluorescence analysis revealed staining of small, newly formed muscle fibers in regenerating but not normal adult muscle cells. By using of fluorescence in situ hybridization, the ADAM12 gene is mapped to human chromosome 10q26.3.

Kit Components

Catalog Number Description Quantity
EK1293-CAP Anti-Rat ADAM12 Pre-coated 96-well strip microplate 1
EK1293-ST Rat ADAM12 Standard 2 vials, 10 ng/tube
EK1293-DA Rat ADAM12 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)015631262512500025000500010,000
O.D.0.0330.0550.1110.2070.3990.7351.2512.029

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect ADAM12, Dilution ratio of 1:1, concentration in serum and plasma is 320-520 pg/ml..

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Rat ADAM12 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)2171804626123718496482
Standard deviation9.54137.1306.7810.9144.22388.92
CV (%)4.4%7.6%4.9%4.6%7.8%6%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of ADAM12 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 121718521221520712.966.2%
Sample 21804173918651710177959.963.3%
Sample 362615528617759995991283.694.7%
*number of samples for each test n=16.

Gene/Protein Information For ADAM12 (Source: Uniprot.Org, NCBI)

Gene Name

ADAM12

Full Name

Disintegrin and metalloproteinase domain-containing protein 12

Weight

99.542kDa

Alternative Names

a disintegrin and metalloproteinase domain 12 (meltrin alpha); ADAM 12; ADAM metallopeptidase domain 12; ADAM12; disintegrin and metalloproteinase domain-containing protein 12; EC 3.4.24; EC 3.4.24.58; MCMP; MCMPMltna; Meltrin alpha; meltrin-alpha; MLTNA; MLTNEC 3.4.24.- ADAM12 ADAM12-OT1, CAR10, MCMP, MCMPMltna, MLTN, MLTNA ADAM metallopeptidase domain 12 disintegrin and metalloproteinase domain-containing protein 12|meltrin-alpha|metalloprotease-disintegrin 12 transmembrane

*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 ADAM12, check out the ADAM12 Infographic

ADAM12 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 ADAM12: 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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11 Customer Q&As for Rat ADAM12 ELISA Kit PicoKine®

Question

Q: we need your recommendation regarding the dilution ratio of serum samples for detection of ADAM12 in Rat plasma? I am trying to measure a few parameters and it requires 100ul of diluted samples for each well. We have limited sample volumes so we like to dilute as much as possible.

T. Varadkar

Verified customer

Asked: 2020-06-20

Answer

A: having little idea about 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 ADAM12 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: 2020-06-20

Question

Q: if the enzyme conjugated ADAM12 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 ADAM12 antigen?

B. Gandhi

Verified customer

Asked: 2019-12-11

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: 2019-12-11

Question

Q: how to thaw whole blood sample for ADAM12 ELISA after freezing?

G. Walker

Verified customer

Asked: 2019-01-26

Answer

A: do not freeze and thaw whole blood. erythrocytes are fragile and, if frozen and thawed, will undergo hemolysis rendering the samples useless. To keep your blood samples to test ADAM12 for a later time, you should let the blood clot in glass tubes and separate the serum to freeze for later analysis.

Boster Scientific Support

Answered: 2019-01-26

Question

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

Verified Customer

Verified customer

Asked: 2019-01-13

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 depends upon the kit used so please refer to each datasheet for details.

Boster Scientific Support

Answered: 2019-01-13

Question

Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for measuring ADAM12 using the EK1293 Rat ADAM12 Picokine® ELISA Kit?

Verified Customer

Verified customer

Asked: 2019-01-04

Answer

A: Boster Bio does not assess sample stability. Variations in sample collection, processing, and storage may affect the stabilityof samples. It is recommend to assay sample right after collection when possible, or aliquot into single use volumes and store samples frozen. avoid repetitive freeze-thaw cycles with the stored samples in order to prevent protein degradation.

Boster Scientific Support

Answered: 2019-01-04

Question

Q: can I use citrate plasma as samples in Rat ADAM12 Picokine® ELISA Kit (Catalog # EK1293)?

V. Jha

Verified customer

Asked: 2018-07-30

Answer

A: Chelating agents such as EDTA, Heparin and Citrate can bind metal ions from the functional domain of ADAM12 causing degradation of its protein structure. ADAM12 may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes a must for detection and block the antigen antibody reaction. We have tested the ADAM12 ELISA, treating samples with different anticoagulants and decided that heparin can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.

Boster Scientific Support

Answered: 2018-07-30

Question

Q: Can ADAM12 ELISA Kits be used with tissue homogenates (or other non-validated sample types)?

W. Smith

Verified customer

Asked: 2018-06-02

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 required. 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 evaluated 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: 2018-06-02

Question

Q: what is the protocol regarding preparation of cell lysates prepared for use in Picokine® ELISA kits?

Verified Customer

Verified customer

Asked: 2018-03-30

Answer

A: in those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate are present in the product insert. Components in lysate and lysis buffer may impact immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.

Boster Scientific Support

Answered: 2018-03-30

Question

Q: Are Boster Bio recombinant proteins and antibodies sterile?

W. Dhawan

Verified customer

Asked: 2018-02-12

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 required for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.

Boster Scientific Support

Answered: 2018-02-12

Question

Q: how to proceed with the analysis of ELISA data? I have performed ADAM12 level in plasma.

Verified Customer

Verified customer

Asked: 2016-06-03

Answer

A: please read this article on ELISA data analysis. bosterbio.com/ELISA-data-analysis-instructions. Boster also provides a free tool that you can use to analyze ELISA data. bosterbio.com/biology-research-tools/ELISA-data-analysis-online

Boster Scientific Support

Answered: 2016-06-03

Question

Q: What is the optimal O.D. value for ADAM12 ELISA kit? I performed your ADAM12 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. is it safe to say these samples contain ADAM12 even though the O.D. values are not very high?

G. Kulkarni

Verified customer

Asked: 2016-04-27

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. what you should focus on 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 proportion 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 converting 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 ADAM12 ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of ADAM12 that can be declared/interpreted as positive by the above standard is 10pg/ml.

Boster Scientific Support

Answered: 2016-04-27

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