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Human Agrin (C-terminal fragment) ELISA Kit (A87452)

1 Citation
Standard Curve - Human Agrin (C-terminal fragment) ELISA Kit (A87452) - Antibodies.com Linearity - Human Agrin (C-terminal fragment) ELISA Kit (A87452) - Antibodies.com ELISA - Human Agrin (C-terminal fragment) ELISA Kit - Antibodies.com
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Shipping Information

$40
Dispatched from St. Louis, MO.
Lead Time: 7-11 business days.
Name
Human Agrin (C-terminal fragment) ELISA Kit
Description
Human Agrin (C-terminal fragment) ELISA Kit is a 4 hour sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human Agrin (C-terminal fragment) in serum, plasma, tissue homogenates, and other biological fluids.
Assay Type
Sandwich (quantitative)
Principle of Assay
Human Agrin (C-terminal fragment) ELISA Kit (A87452) is a 4 hour sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human Agrin (C-terminal fragment) in serum, plasma, tissue homogenates, and other biological fluids. A 96-well microtiter plate has been pre-coated with an antibody specific for Agrin (C-terminal fragment). Standards and samples are added to the wells, where Agrin (C-terminal fragment) binds to the immobilized capture antibody. Following incubation, unbound components are removed by washing and a biotinylated detection antibody specific for Agrin (C-terminal fragment) is added to the wells. The detection antibody binds the captured Agrin (C-terminal fragment) to form an antibody-antigen-antibody sandwich complex. After further incubation and washing, HRP-Streptavidin conjugate is added to the wells. TMB substrate solution is then added to visualize the HRP enzymatic reaction. This produces a blue-coloured product that changes to yellow following addition of acidic stop solution. The intensity of the yellow colour is directly proportional to the amount of Agrin (C-terminal fragment) captured in each well. The concentration of Agrin (C-terminal fragment) is calculated by reading the absorbance at 450 nm and comparing the values with the standard curve.
Platform
Pre-coated Microplate (12 x 8 well strips)
Detection Type
Colorimetric Detection (OD450)
Instrument
Colorimetric Microplate Reader
Sample Type
Serum, plasma, tissue homogenates, and other biological fluids.
Sensitivity
28.125 pg/mL
Product Range
46.875-3000 pg/mL
Assay Time
4 hours
Reactivity
Human
Cross Reactivity
Specific for Agrin (C-terminal fragment) with no reported cross-reactivity with other analogs or variants.
Recovery
Sample Type n Range Average
Serum 10 92% - 104% 92%
EDTA Plasma 10 92% - 100% 93%
Heparin Plasma 10 90% - 105% 97%
Linearity
Sample Type n 1:2 1:4 1:8
Serum 10 88-102% 85-104% 86-102%
EDTA Plasma 10 86-98% 85-101% 82-96%
Heparin Plasma 10 83-97% 85-100% 80-95%
Storage
Shipped at +4°C. Store kit components at +4°C or -20°C as instructed in the Protocol Manual. Do not use past expiration date! Avoid freeze/thaw cycles.
Product Note
Information online is representative. Always refer to the Product Manual inside the kit.
Disclaimer
This product is for research use only. It is not intended for diagnostic or therapeutic use.
Kit Components
Item Quantity Storage
Pre-Coated 96 Well Microplate 12 x 8 Well Strips +4°C
Lyopholized Standard 2 Vials +4°C
Sample Dilution Buffer 20ml +4°C
Biotinylated Detection Antibody 120µl +4°C
Antibody Dilution Buffer 10ml +4°C
HRP-Streptavidin Conjugate 120µl +4°C
SABC Dilution Buffer 10ml +4°C
TMB Substrate 10ml +4°C
Stop Solution 10ml +4°C
Wash Buffer (25X) 30ml +4°C
Plate Sealers 5 Adhesive Strips -
Foil Pouch 1 Zip-Sealed Pouch -
Required Materials
The following materials are not included in the kit, but are required to run this assay:
  • Microplate reader capable of measuring absorbance at 450nm ± 10nm.
  • Squirt bottle, multichannel pipette reservoir, or automated microplate washer.
  • Graph paper or computer software capable of generating logarithmic functions.
  • Test tubes and Eppendorf tubes to prepare standards and samples.
  • Multi- and single-channel pipettes and sterile pipette tips.
  • Deionized or distilled water.
  • Absorbent paper towels.
  • 37°C incubator.
Important Notes
  • Sample concentrations should be estimated before being used in the assay and a proper dilution factor should be selected to make the diluted target protein concentration fall in the optimal detection range of this kit.
  • Sample matrix components will interfere with the test results. All samples must be diluted at least 1:2 with Sample Dilution Buffer.
  • Hemolysis can greatly impact the validity of test results. Take care to minimize hemolysis.
  • This ELISA kit is intended for the analysis of biological samples and has not been validated for purified/recombinant proteins, industrial, or commercial materials. Performance with such samples cannot be guaranteed, as differences in expression, sequence, tags, folding, post-translational modifications, purity, activity, or buffer composition may affect assay results. For further guidance, please contact our technical support team.
Bring all reagents and samples to room temperature before use.
We recommend that all standards and samples be assayed in duplicate.
Step 1
Prepare all reagents, samples, and working standards as directed in the previous sections.
Step 2
Determine the number of microplate strips required for the assay and insert them into the plate frame. Unused strips should be stored in the foil pouch at +4°C.
Step 3
Add 100μl of standard solutions to the standard wells. Note: A plate layout is provided to record standards and samples assayed.
Step 4
Add 100μl of samples to the sample wells. Note: All samples must be properly diluted in order to produce sample values within the dynamic range of the assay. All samples must be diluted at least 1:2 with Sample Dilution Buffer.
Step 5
Cover with an adhesive plate sealer provided and incubate at 37°C for 90 minutes.
Step 6
Remove the plate sealer, aspirate the liquid from the plate, and wash the plate two times with Wash Buffer. Soak wells with 350μl of Wash Buffer for 1-2 minutes each time. Note: Complete removal of liquid at each step is essential to good performance. After the last wash, remove any remaining Wash Buffer by aspirating or decanting. Blot the plate onto paper towels or other absorbent material. Do not let the wells dry completely at any time!
Step 7
Add 100μl of Biotinylated Detection Antibody working solution to each well.
Step 8
Cover with a new adhesive plate sealer and incubate at 37°C for 60 minutes.
Step 9
Remove the plate sealer, aspirate the liquid from the plate, and wash the plate three times with Wash Buffer. Soak wells with 350μl of Wash Buffer for 1-2 minutes each time.
Step 10
Add 100μl of HRP-Streptavidin Conjugate (SABC) working solution to each well.
Step 11
Cover with a new adhesive plate sealer and incubate at 37°C for 30 minutes.
Step 12
Remove the plate sealer, aspirate the liquid from the plate, and wash the plate five times with Wash Buffer. Soak wells with 350μl of Wash Buffer for 1-2 minutes each time.
Step 13
Add 90μl of TMB Substrate to each well. Cover with a new adhesive plate sealer and incubate at 37°C in the dark for 10-20 minutes. Note: This incubation time is for reference only, the optimal reaction time should be determined by the end-user and can be shortened or extended according to the actual color change. The reaction may be terminated when a gradient is apparent in the standard wells. Do not exceed 30 minutes!
Step 14
Add 50μl of Stop Solution to each well. The color in the wells should change from blue to yellow immediately. Note: If the color in the wells is green or the color change does not appear uniform, gently tap the plate to ensure thorough mixing.
Step 15
Determine the optical density (O.D. value) of each well immediately using a microplate reader set to 450 nm.

  • Scientific Validation DataValidation Data (3)

Standard Curve - Human Agrin (C-terminal fragment) ELISA Kit (A87452) - Antibodies.comEnlarge Image

Standard Curve - Human Agrin (C-terminal fragment) ELISA Kit (A87452)

Representative standard curve for Human Agrin (C-terminal fragment) ELISA Kit (A87452).
Linearity - Human Agrin (C-terminal fragment) ELISA Kit (A87452) - Antibodies.comEnlarge Image

Human Agrin (C-terminal fragment) ELISA Kit (A87452)

Linearity assessment of the Human Agrin (C-terminal fragment) ELISA Kit (A87452) across serial sample dilutions. Linearity was evaluated using Serum, EDTA Plasma, and Heparin Plasma samples serially diluted from 1:2 to 1:8. Recovery (%) was calculated relative to the expected Human Agrin (C-terminal fragment) concentration at each dilution. Data are presented as mean recovery values from 10 replicates (n = 10).
ELISA - Human Agrin (C-terminal fragment) ELISA Kit - Antibodies.comEnlarge Image

Human Agrin (C-terminal fragment) ELISA Kit (A87452)

ELISA test using serum concentrations of C-terminal agrin fragment (CAF) measured pre (day 0) and post (day 5) 5 days of bed rest with concurrent unilateral neuromuscular electrical stimulation in young (circles) and old (triangles) adults. Data are presented as mean ± SD with individual data points shown. No significant changes were observed over time.
Figure 3B from Hansen SK et al. (2025). Originally published in Exp Physiol, DOI: 10.1113/EP092194. Epub 2025 Mar 31.. Reproduced under CC BY 4.0.

  • Product Citations (1)

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Effects of neuromuscular electrical stimulation on voluntary muscle activation and peripheral muscle contractility following short-term bed rest
This publication cites Human Agrin (C-terminal fragment) ELISA Kit (A87452).
Article Abstract

Disuse induces a disproportionate loss of muscle force compared with muscle mass, with unclear effects on voluntary muscle activation (VA) and peripheral contractility. Furthermore, the effect of neuromuscular electrical stimulation (NMES) as a disuse countermeasure remains uncertain. We investigated the effects of NMES during bed rest on neuromechanical function to improve our understanding of the mechanisms underlying disuse-induced reductions in muscular force. Young (n = 16, 25 years old) and old (n = 16, 71 years old) adults underwent 5 days of bed rest. One leg received NMES (3 × 30 min/day), while the other served as the control (CON). Maximal isometric knee-extensor strength (MVIC), VA and peripheral muscle contractility were assessed before and after bed rest using the interpolated twitch technique, along with biomarkers of neuromuscular junction instability (C-terminal agrin fragment (CAF)) and muscle damage (creatine kinase (CK)). MVIC decreased in both age groups, regardless of NMES (young: CON, -21.7 Nm and NMES, -23.8 Nm; old: CON, -18.5 Nm and NMES, -16.4 Nm). VA was preserved with NMES, while decreasing in CON legs (young, -8.1%; old, -5.6%) following bed rest. Peripheral contractility (resting doublet twitch force) was reduced in CON and NMES legs in both age groups (young: CON, -4.0 Nm and NMES, -11.5 Nm; old: CON, -5.9 Nm and NMES, -10.8 Nm), with a greater decrease in NMES legs. CAF remained unchanged, whereas CK levels increased in young participants, albeit remaining within the normal range. In conclusion, a decline in neuromechanical function was observed after 5 days of bed rest in young and old adults. Although NMES appeared to preserve VA, peripheral muscle contractility was altered, resulting in reduced MVIC.

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