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Mouse/Rat IGF-I/IGF-1 ELISA Kit - Quantikine Best Seller

R&D Systems, part of Bio-Techne | Catalog # MG100

R&D Systems, part of Bio-Techne
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MG100
PMG100
SMG100

Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (25 µL), Tissue Homogenates (50 µL), Serum (10 µL), EDTA Plasma (10 µL), Heparin Plasma (10 µL)

Sensitivity

8.4 pg/mL

Assay Range

31.2-2000 pg/mL (Cell Culture Supernates, Tissue Homogenates, Serum, EDTA Plasma, Heparin Plasma)

Product Summary for Mouse/Rat IGF-I/IGF-1 Quantikine ELISA Kit

The Quantikine Mouse/Rat IGF-I Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse and rat IGF-I in cell culture supernates, tissue homogenates, serum, and plasma. It contains E. coli-expressed recombinant mouse IGF-I and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse and rat IGF-I showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring mouse/rat IGF-I.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Reactivity

Mouse, Rat

Specificity

Natural and recombinant mouse and rat IGF-I

Cross-reactivity

< 0.5% cross-reactivity observed with available related molecules. < 50% cross-species reactivity observed with species tested.

Interference

No significant interference observed with available related molecules.

Sample Values

Serum/Plasma - Individual mouse and rat serum and plasma samples were evaluated for detectable levels of mouse and rat IGF-1 in this assay. Serum and plasma samples were not matched.

Sample TypeMean (ng/mL)Range (ng/mL)Standard Deviation (ng/mL)
Mouse serum (n=20)742513-924113
Mouse EDTA plasma (n=20)593346-806130
Mouse heparin plasma (n=20)821542-1447203

Sample TypeMean (ng/mL)Range (ng/mL)Standard Deviation (ng/mL)
Rat serum (n=20)1158436-1908376
Rat EDTA plasma (n=20)1016430-2207377
Rat heparin plasma (n=20)1012723-1444208

Cell Culture Supernates:
J774A.1 mouse reticulum cell sarcoma macrophage cells (1 x 106 cells/mL) were cultured for 3 days in DMEM containing 10% fetal bovine serum and stimulated with 2.5 ng/mL LPS. The cell culture supernate was assayed for mouse IGF-1 and measured 5842 pg/mL.

WEHI-3 mouse myelomonocytic leukemia cells were cultured for 2-4 days in RPMI supplemented with 10% fetal bovine serum. The cell culture supernate was assayed for mouse IGF-1 and measured 4944 pg/mL.

Rat lung conditioned media (1 lung, 1-2 mm pieces) was cultured for 2 days in 50 mL of DMEM supplemented with 10% fetal bovine serum and stimulated with 100 ng/mL LPS and 50 ng/mL PMA. The cell culture supernate was assayed for rat IGF-1 and measured 1514 pg/mL.

Rat heart conditioned media (1 heart, 1-2 mm pieces) was cultured for 4 days in 30 mL of RPMI, supplemented with 10% fetal bovine serum, and stimulated with 2.5 ng/mL LPS. The cell culture supernate was assayed for rat IGF-1 and measured 2888 pg/mL.

Tissue Homogenates:
Homogenates from mouse lung and mouse heart tissue were assayed for levels of mouse IGF-1 and measured 5862 pg/mL and 3158 pg/mL, respectively. 
Homogenates from rat lung and rat heart tissue were assayed for levels of rat IGF-1 and measured 5476 pg/mL and 1785 pg/mL, respectively.


Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision.

Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision.

Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Serum, Tissue Homogenates

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 35 35 35
Mean (pg/mL) 82 269 921 86 256 863
Standard Deviation 4.6 8.9 37.7 7.8 11.8 37.4
CV% 5.6 3.3 4.1 9.1 4.6 4.3

Recovery for Mouse/Rat IGF-I/IGF-1 Quantikine ELISA Kit

The recovery of mouse/rat IGF-I spiked to three levels throughout the range of the assay in the cell culture supernate samples was evaluated.

Sample Type Average % Recovery Range %
Mouse Cell Culture Supernates (n=7) 98 89-111
Rat Cell Culture Supernates (n=6) 98 82-108

Linearity

To assess the linearity of the assay, samples containing mouse or rat IGF-I in each matrix were diluted with Calibrator Diluent and then assayed.

Mouse/Rat IGF-I ELISA Linearity
Mouse/Rat IGF-I ELISA Linearity

Scientific Data Images for Mouse/Rat IGF-I/IGF-1 Quantikine ELISA Kit

Mouse/Rat IGF-I ELISA Standard Curve

Mouse/Rat IGF-I ELISA Standard Curve

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: IGF-I/IGF-1

Insulin-like Growth Factor 1 (IGF-1), also known as somatomedin C, is a member of the insulin superfamily. It was originally discovered as a mediator of growth hormone actions on somatic cell growth, but has also been shown to be an important regulator of cell metabolism, differentiation and survival. IGF-1 is synthesized as a preproprotein that is proteolytically cleaved to generate the mature protein linked by three disulfide bonds. Mature IGF-1 is highly conserved among large mammals, with 100% sequence identity between the human, bovine, porcine, equine, and canine proteins. 

Mature mouse IGF-1 is a non-glycosylated, 70 amino acid (aa) secreted polypeptide that is derived from either a 153 aa or a 159 aa preproprotein. It shares 99% and 94% aa sequence identity with rat and human IGF-1, respectively. IGF-1 is synthesized in the liver and other tissues. It is found in blood and other body fluids as a complex with specific high affinity IGF binding proteins (IGFBP-1 to -6). The IGFBPs are expressed in specific patterns during development. They are modulators of IGF actions, which control IGF bioavailability to specific cell-surface receptors. Their functions are further regulated by IGFBP proteases, which proteolytically cleave the IGFBPs to lower the affinity with which they bind IGFs and increase IGF bioavailability. Some IGFBPs also have IGF-Independent effects on cell functions. IGF-Independent circulates primarily as a ternary complex with IGFBP-3 or IGFBP-5 and the acid-labile subunit (ALS). Some IGF-1 is also present in binary complexes with other IGFBPs. Whereas the ternary complexes are generally restricted to the vasculature, the binary complexes freely enter the tissues. 

IGF-1 actions are mediated by two ubiquitously expressed receptor tyrosine kinases: IGF-I R and Insulin R/CD220. IGF-I R and Insulin R are disulfide-linked heterotetrameric complexes that consist of two alpha and two beta subunits. For both of these receptors, the prepro proteins are cleaved to produce extracellular alpha subunits which contain a cysteine-rich region and ligand-binding fibronectin type III (FN-III) domains, and beta subunits which contain an extracellular FN-III domain, transmembrane, and cytoplasmic tyrosine kinase domains. A hybrid complex containing one IGF-I R and one Insulin R also serves as a functional high affinity receptor for IGF-1. IGF-I R-Insulin R hybrids respond primarily to IGF-1, potentially downregulating the cellular response to Insulin. IGF signaling is also modulated by IGF binding proteins and the scavenger receptor, IGF-I I R.

Long Name

Insulin-like Growth Factor I/Insulin-like Growth Factor 1

Alternate Names

IGF-1, IGF1, IGFI, Somatomedin A, Somatomedin C

Entrez Gene IDs

3479 (Human); 16000 (Mouse); 24482 (Rat)

Gene Symbol

IGF1

Additional IGF-I/IGF-1 Products

Product Documents for Mouse/Rat IGF-I/IGF-1 Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Mouse/Rat IGF-I/IGF-1 Quantikine ELISA Kit

For research use only

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⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.