Human Free IGFBP-1 Quantikine ELISA Kit

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DGB100
Control Products Available
Human Free IGFBP-1 ELISA Standard Curve
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Product Details
Procedure
Citations (5)
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Human Free IGFBP-1 Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Cell Lysates (50 uL), Serum (10 uL), EDTA Plasma (10 uL), Heparin Plasma (10 uL), Urine (10 uL), Human Milk (10 uL)
Sensitivity
13.8 pg/mL
Assay Range
125.0 - 8,000 pg/mL (Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine, Human Milk)
Specificity
Natural and recombinant phosphorylated and non-phosphorylated human free IGFBP-1. This assay does not detect IGFBP-1 in complex with IGF-I or IGF-II.
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.

Product Summary

The Quantikine Human Free IGFBP-1 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human free IGFBP-1 in cell culture supernates, cell lysates, serum, plasma, urine, and human milk. It contains NS0-expressed recombinant human IGFBP-1 and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human IGFBP-1 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 human free IGFBP-1.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components

Cell Culture Supernates, Cell Lysates, Serum, EDTA Plasma, Heparin Plasma, Urine, Human Milk

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pg/mL) 1038 3065 5639 1079 3100 5895
Standard Deviation 58.2 131 230 103 209 373
CV% 5.6 4.3 4.1 9.5 6.7 6.3

Recovery

The recovery of human Free IGFBP-1 spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 112 92-117
Cell Lysates (n=4) 107 91-117
EDTA Plasma (n=4) 97 86-111
Heparin Plasma (n=4) 97 85-108
Human Milk (n=4) 104 92-114
Serum (n=4) 94 86-107

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human Free IGFBP-1 were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Free IGFBP-1 ELISA Linearity

Scientific Data

Human Free IGFBP-1 ELISA Standard Curve

Product Datasheets

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Preparation and Storage

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

Background: IGFBP-1

The superfamily of insulin-like growth factor (IGF) binding proteins include the six high-affinity IGF binding proteins (IGFBP) and at least four additional low-affinity binding proteins referred to as IGFBP related proteins (IGFBP-rP). All IGFBP superfamily members are cysteine-rich proteins with conserved cysteine residues, which are clustered in the amino- and carboxy-terminal thirds of the molecule. IGFBPs modulate the biological activities of IGF proteins. Some IGFBPs may also have intrinsic bioactivity that is independent of their ability to bind IGF proteins. Post-translational modifications of IGFBP, including glycosylation, phosphorylation and proteolysis, have been shown to modify the affinities of the binding proteins to IGF. ALS (Acid Labile Subunit) is a liver-derived protein that exists in a ternary complex with Insulin-like Growth Factor (IGF)-binding Protein-3 (IGFBP-3) or IGFBP-5, and either IGF-I or IGF-II. ALS increases the half-life of IGF/IGFBP complexes in circulation.

Long Name:
Insulin-like Growth Factor Binding Protein 1
Entrez Gene IDs:
3484 (Human); 16006 (Mouse)
Alternate Names:
alpha-pregnancy-associated endometrial globulin; amniotic fluid binding protein; binding protein-25; binding protein-26; binding protein-28; growth hormone independent-binding protein; hIGFBP-1; IBP1; IBP-1; IGF-binding protein 1; IGFBP1; IGFBP-1; IGF-BP25; insulin-like growth factor binding protein 1; insulin-like growth factor-binding protein 1; Placental protein 12; PP12AFBP
&#9888; 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.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 50 µL Assay Diluent
  4.   Add 50 µL of Assay Diluent to each well.

  5. 50 µL Standard, Control, or Sample
  6.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at 2-8 °C for 3 hours.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.

Citations for Human Free IGFBP-1 Quantikine ELISA Kit

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.

5 Citations: Showing 1 - 5
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  1. Placental RNA sequencing implicates IGFBP1 in insulin sensitivity during pregnancy and in gestational diabetes
    Authors: Hivert, MF;White, F;Allard, C;James, K;Majid, S;Aguet, F;Ardlie, K;Edlow, A;Florez, J;Bouchard, L;Jacques, PE;Karumanchi, S;Powe, C;
    Research square
    Species: Human
    Sample Types: Plasma
  2. EGF, TGF-? and Amphiregulin Differently Regulate Endometrium-Derived Mesenchymal Stromal/Stem Cells
    Authors: Kamentseva, RS;Kharchenko, MV;Gabdrahmanova, GV;Kotov, MA;Kosheverova, VV;Kornilova, ES;
    International journal of molecular sciences
    Species: Human
    Sample Types: Cell Culture Supernates
  3. The fate of human SUSD2+ endometrial mesenchymal stem cells during decidualization
    Authors: T Gorsek Spa, UR Markert, P Reif, W Schoell, G Moser, J Feichtinge, ZN Mihalic, J Kargl, CE Gargett, D Gold
    Stem Cell Research, 2022-01-15;60(0):102671.
    Species: Human
    Sample Types: Cell Culture Supernates
  4. Human umbilical cord blood mesenchymal stem cells engineered to overexpress growth factors accelerate outcomes in hair growth
    Authors: DH Bak, MJ Choi, SR Kim, BC Lee, JM Kim, ES Jeon, W Oh, ES Lim, BC Park, MJ Kim, J Na, BJ Kim
    Korean J. Physiol. Pharmacol., 2018-08-27;22(5):555-566.
    Species: Human
    Sample Types: Cell Culture Supernates
  5. Chronic endometritis modifies decidualization in human endometrial stromal cells
    Authors: D Wu, F Kimura, L Zheng, M Ishida, Y Niwa, K Hirata, A Takebayash, A Takashima, K Takahashi, R Kushima, G Zhang, T Murakami
    Reprod. Biol. Endocrinol, 2017-03-04;15(1):16.
    Species: Human
    Sample Types: Cell Culture Supernates

FAQs

  1. Assay Diluent RD1-14 contains visible precipitate. Is this acceptable to use?

    • A small amount of precipitate is expected and acceptable in Assay Diluent RD1-14. Variability has been observed between vials and diluent lots. It may not fully resolubilize but this will not impact assay performance. It is not recommended to filter the precipitate.

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