Human Fetuin A/AHSG ELISA Kit PicoKine®

AHSG ELISA kit for Human

Human Fetuin A / AHSG ELISA Kit PicoKine™ (96 Tests). Quantitate Human AHSG in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 10pg/ml. Cited in 7 publication(s).

Product Info Summary

SKU: EK0757
Size: 96 wells/kit, with removable strips.
Reactive Species: Human
Application: ELISA
Sample Types: cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Product Name

Human Fetuin A/AHSG ELISA Kit PicoKine®

View all AHSG ELISA kits

SKU/Catalog Number

EK0757

Size

96 wells/kit, with removable strips.

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

Description

Human Fetuin A / AHSG ELISA Kit PicoKine™ (96 Tests). Quantitate Human AHSG in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). 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

Human Fetuin A/AHSG ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK0757)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: NS0; Immunogen sequence: A19-V367

Sensitivity

<10 pg/ml

Assay Range

0.78 ng/ml - 50 ng/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 EK0757 for more details

Cross-reactivity

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

Reactive Species

EK0757 is reactive to AHSG in Human samples

Validated Sample Types

cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Application Guarantee

EK0757 is guaranteed for ELISA in Human by Boster Guarantee

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

Background of AHSG

Alpha2-HS glycoprotein (AHSG), also known as fetuin-A, is a plasma protein displaying high-affinity interaction with calcium phosphate, by which ectopic vascular calcification is prevented. The AHSG polymorphism is attributable to the hereditary variation of AHSG and phosphate serum levels, which may affect skeletal development and chronic disorders such as vascular calcification.1Human plasma protein alpha 2-HS-glycoprotein (AHSG) is composed of two polypeptide chains, A and B, encoded by a single mRNA. Southern blot analysis of mouse x human somatic cell hybrids has mapped the AHSG gene to human chromosome 3 in the region 3q21----qter. Using a recombinant plasmid containing a 1,538 bp insert spanning the entire AHSG coding region, AHSG was localized to chromosomal bands 3q27----q29 by in situ hybridization.2

Kit Components

Catalog Number Description Quantity
EK0757-CAP Anti-Human AHSG Pre-coated 96-well strip microplate 1
EK0757-ST Human AHSG Standard 2 vials, 50 ng/tube
EK0757-DA Human AHSG 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 (ng/ml)00.781.563.126.2512.52550
O.D.0.0710.3060.3960.6731.1691.5851.7192.026

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect AHSG, Dilution ratio of 1:100000, concentration in serum and plasma is 219-805 ug/ml..

Some articles we found to cite concentrations of AHSG in samples: 25279156 (Pubmed IDs).

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Human Fetuin A/AHSG 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)16464628220951708502121195
Standard deviation126.74185.82883.8143.4722441229.31
CV (%)7.7%4%4%8.4%4.5%5.8%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of AHSG 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 11646172617661528166690.885.4%
Sample 24628449546034388452895.272.1%
Sample 322095251022261325109237291387.885.8%
*number of samples for each test n=16.

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

Gene Name

AHSG

Full Name

Alpha-2-HS-glycoprotein

Weight

39.341kDa

Superfamily

fetuin family

Alternative Names

A2HS; AHS; AHSG; alpha-2-HS-glycoprotein; alpha-2-Z-globulin; Ba-alpha-2-glycoprotein; FETUAba-alpha-2-glycoprotein; Fetuin A; fetuin-A; HSGA AHSG A2HS, AHS, APMR1, FETUA, HSGA alpha 2-HS glycoprotein alpha-2-HS-glycoprotein|alpha-2-Z-globulin|ba-alpha-2-glycoprotein|fetuin-A

*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 AHSG, check out the AHSG Infographic

AHSG 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 AHSG: 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].

Hello CJ!

EK0757 has been cited in 7 publications:

*The publications in this section are manually curated by our staff scientists. They may differ from Bioz's machine gathered results. Both are accurate. If you find a publication citing this product but is missing from this list, please let us know we will issue you a thank-you coupon.

Plasma fetuin-A/α2-HS-glycoprotein correlates negatively with inflammatory cytokines, chemokines and activation biomarkers in individuals with type-2 diabetes

Cardiometabolic risks vary by weight status in pediatric kidney and liver transplant recipients: A cross‐sectional, single‐center study in the USA

The relationship of pentraxin-3 levels with IL-17, fetuin-A, insulin in patients with Behçets disease

Levels of serum pentraxin 3, IL-6, fetuin A and insulin in patients with rheumatoid arthritis

Plasma fetuin-A/?2-HS-glycoprotein correlates negatively with inflammatory cytokines, chemokines and activation biomarkers in individuals with type-2 diabetes

He, S., Le, N.A., Frediani, J.K., Winterberg, P.D., Jin, R., Liverman, R.,..., & Vos, M.B. (2017). Cardiometabolic risks vary by weight status in pediatric kidney and liver transplant recipients: A cross-sectional, single-center study in the USA. ...

Kocyigit I, Unal A, Elcik D, Korkar H, Sen A, Yasan M, Eroglu E, Sipahioglu Mh, Tokgoz B, Oymak O. Transplant Proc. 2015 Jun;47(5):1398-401. Doi: 10.1016/J.Transproceed.2015.04.008. Association Between Cardiac Valvular Calcification And Serum Fetu...

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10 Customer Q&As for Human Fetuin A/AHSG ELISA Kit PicoKine®

Question

Q: can you recommend the dilution ratio of serum samples for detection of AHSG in Human serum? I am trying to measure a a number of analytes and it requires 100ul of diluted samples for each well. We have low serum quantitys so we like to dilute as much as possible.

S. Liu

Verified customer

Asked: 2020-11-28

Answer

A: without knowing 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 AHSG 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: 2020-11-28

Question

Q: if the enzyme conjugated AHSG 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 AHSG antigen?

C. Dhawan

Verified customer

Asked: 2020-09-22

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: 2020-09-22

Question

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

Verified Customer

Verified customer

Asked: 2020-08-29

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 suitable 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: 2020-08-29

Question

Q: What is the optimal O.D. value for AHSG ELISA kit? I performed your AHSG 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 AHSG even though the O.D. values are not very high?

Verified Customer

Verified customer

Asked: 2020-06-24

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 your example, 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 relation 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 AHSG ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of AHSG that can be declared/interpreted as positive by the above standard is 10pg/ml.

Boster Scientific Support

Answered: 2020-06-24

Question

Q: how can I thaw whole blood sample for AHSG ELISA after freezing?

Verified Customer

Verified customer

Asked: 2020-04-21

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 AHSG 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: 2020-04-21

Question

Q: can you tell me how to prepare cell lysates prepared for use in Picokine® ELISA kits?

C. Lewis

Verified customer

Asked: 2020-04-09

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 can have an affect on immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.

Boster Scientific Support

Answered: 2020-04-09

Question

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

Verified Customer

Verified customer

Asked: 2020-02-17

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 might differ by kit so please refer to each datasheet for details.

Boster Scientific Support

Answered: 2020-02-17

Question

Q: for how much duration can samples (cell cultures, serum, and plasma) be stored and still be stable for measurement AHSG using the EK0757 Human AHSG Picokine® ELISA Kit?

P. Maharaj

Verified customer

Asked: 2020-02-14

Answer

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

Boster Scientific Support

Answered: 2020-02-14

Question

Q: is there any online tool I can use to streamline the data analysis for my ELISA results?

B. Zhou

Verified customer

Asked: 2019-08-09

Answer

A: We have a web based ELISA curve fitting (4pl) and data analysis tool. Please give it a try: bosterbio.com/biology-research-tools/ELISA-data-analysis-online. You can also consult our article on ELISA data analysis: bosterbio.com/ELISA-data-analysis-instructions

Boster Scientific Support

Answered: 2019-08-09

Question

Q: is it okay to use citrate plasma as samples in Human AHSG Picokine® ELISA Kit (Catalog # EK0757)?

Verified Customer

Verified customer

Asked: 2019-04-21

Answer

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

Boster Scientific Support

Answered: 2019-04-21

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