Human SOCS2 ELISA Kit (A77352)

Name: Human SOCS2 ELISA Kit
Brand: Antibodies.com
SKU: A77352
Price: 440.0 GBP
Availability: InStock

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Name

Human SOCS2 ELISA Kit

Description

Human SOCS2 ELISA Kit is a sandwich Enzyme-Linked Immunosorbent Assay (sELISA) designed for the in vitro quantitative determination of human SOCS2 in serum, plasma, tissue homogenates, and other biological fluids.

Assay Type

Sandwich (quantitative)

Principle of Assay

Human SOCS2 ELISA Kit (A77352) employs the sandwich enzyme immunoassay technique for the quantitative measurement of human SOCS2 in serum, plasma, tissue homogenates, and other biological fluids. An antibody specific for SOCS2 has been pre-coated onto a 96-well microtiter plate. The standards and test samples are added into the wells and the SOCS2 present in each sample is bound to the wells by the immobilized antibody. Following incubation, the wells are washed and then incubated with Biotinylated Anti-SOCS2 Antibody, which binds the captured SOCS2 present in each well. Following incubation, unbound biotinylated detection antibody is removed by washing, and an HRP-Streptavidin conjugate is added to the wells and the microtiter plate is incubated. Following incubation and washing, TMB substrate solution is then used to visualize the HRP enzymatic reaction by catalysis to produce a blue-coloured product that changes to yellow after addition of acidic stop solution. The density of yellow is proportional to the amount of SOCS2 captured in each well. The concentration of SOCS2 can then be calculated by reading the O.D. absorbance at 450nm in a microplate reader and referring to the standard curve.

Platform

Pre-coated Microplate (12 x 8 well strips)

Detection Type

Colorimetric

Instrument

Colorimetric Microplate Reader

Sample Type

Serum, plasma, tissue homogenates, and other biological fluids.

Sensitivity

75 pg/ml

Product Range

125-8000 pg/ml

Assay Time

4 hours

Reactivity

Human

Recovery

Sample Type	n	Range	Average
Serum	5	85% - 94%	91%
EDTA Plasma	5	89% - 105%	100%
Heparin Plasma	5	89% - 104%	98%

Linearity

Sample Type	n	1:2	1:4	1:8
Serum	5	85-95%	88-103%	88-101%
EDTA Plasma	5	90-99%	82-96%	82-94%
Heparin Plasma	5	82-99%	83-99%	81-92%

Precision

Intra-assay CV: < 8%, Inter-assay CV: < 10%

Storage

Store at +4°C. Do not use past expiration date!

Synonyms

CIS-2, CIS2, Cytokine-inducible SH2 protein 2, SOCS-2, SSI-2, SSI2, STAT-induced STAT inhibitor 2, STATI2, Suppressor of cytokine signaling 2

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 60 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 (1)

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Standard Curve - Human SOCS2 ELISA Kit (A77352)

Representative standard curve for Human SOCS2 ELISA Kit (A77352).

Product Citations (1)

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Suppressor of cytokine signaling 2 is associated with growth impairment in pediatric chronic kidney disease

This publication cites Human SOCS2 ELISA Kit (A77352).

Article Abstract

Background: Growth failure is a common and serious complication of chronic kidney disease (CKD) in children, resulting from numerous factors, including growth hormone (GH) resistance. Suppressor of cytokine signaling 2 (SOCS2), a negative regulator of GH signaling, has been implicated in growth regulation but has not been previously studied in pediatric CKD.

Methods: In this cross-sectional study, we assessed serum SOCS2 concentrations in 55 children with CKD (stages 2-5) and 27 age- and sex-matched healthy controls using a high-sensitivity ELISA. We evaluated anthropometric parameters, kidney function, and biochemical markers. Associations between SOCS2 levels and growth (height standard deviation scores, HtSDS) were analyzed using correlation and multivariate regression models.

Results: SOCS2 levels were significantly higher in children with CKD compared to controls (median 1526.5 vs. 1294.6 pg/ml, p < 0.001). In patients with CKD, SOCS2 negatively correlated with HtSDS (r = -0.30, p = 0.029). In multivariate analysis, SOCS2 was an independent predictor of lower HtSDS alongside eGFR. SOCS2 concentrations were higher in short-statured children, and ROC analysis showed acceptable diagnostic performance for predicting short stature (AUC 0.78, 95% CI 0.64-0.91; 79.5% sensitivity, 68.8% specificity).

Conclusions: Circulating SOCS2 levels are elevated in pediatric CKD and independently associated with growth impairment, suggesting its potential link to GH resistance, which is typical of CKD. SOCS2 might be a potential new marker of growth retardation. Further longitudinal studies are needed to explore its predictive value for longitudinal growth and GH treatment response.

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