Human AGT/Serpin A8/Angiotensinogen ELISA Kit PicoKine®

AGT ELISA kit for Human

Human AGT/Serpin A8/Angiotensinogen ELISA Kit PicoKine™ (96 Tests). Quantitate Human AGT in cell culture supernatants, serum and plasma (EDTA). Sensitivity: 10pg/ml.

Product Info Summary

SKU: EK1465
Size: 96 wells/kit, with removable strips.
Reactive Species: Human
Application: ELISA
Sample Types: cell culture supernatants, serum and plasma (EDTA).

Product Name

Human AGT/Serpin A8/Angiotensinogen ELISA Kit PicoKine®

View all AGT ELISA kits

SKU/Catalog Number

EK1465

Size

96 wells/kit, with removable strips.

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

Description

Human AGT/Serpin A8/Angiotensinogen ELISA Kit PicoKine™ (96 Tests). Quantitate Human AGT in cell culture supernatants, serum and plasma (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 AGT/Serpin A8/Angiotensinogen ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1465)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: NS0; Immunogen sequence: D34-A485

Sensitivity

<10 pg/ml

Assay Range

312 pg/ml - 20,000 pg/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 EK1465 for more details

Cross-reactivity

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

Reactive Species

EK1465 is reactive to AGT in Human samples

Validated Sample Types

cell culture supernatants, serum and plasma (EDTA).

Application Guarantee

EK1465 is guaranteed for ELISA in Human by Boster Guarantee

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

Background of AGT

AGT (ANGIOTENSINOGEN), also called SERPINA8, is an α-2-globulin that is produced constitutively and released into the circulation mainly by the liver. ATG is a member of the serpin family, although it is not known to inhibit other enzymes, unlike most serpins. Angiotensinogen is also known as renin substrate. The AGT gene is mapped on 1q42.2. And the human angiotensinogen gene contains 5 exons. The expression of AGT and enzymes required for its conversion to angiotensin II in human adipose tissue. Mutations in this AGT gene are associated with susceptibility to essential hypertension, and can cause renal tubular dysgenesis, a severe disorder of renal tubular development. Defects in this gene have also been associated with non-familial structural atrial fibrillation, and inflammatory bowel disease.

Kit Components

Catalog Number Description Quantity
EK1465-CAP Anti-Human AGT Pre-coated 96-well strip microplate 1
EK1465-ST Human AGT Standard 2 vials, 20 ng/tube
EK1465-DA Human AGT 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 (pg/ml)03126251250250050001000020000
O.D.0.0380.1410.2250.3880.7051.0561.7452.060

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect AGT, Dilution ratio of 1:500, concentration in serum and plasma is 1-2 ug/ml..

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Human AGT/Serpin A8/Angiotensinogen 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)3642379737041724418736
Standard deviation18.93126.09405.3525.44156.22585.31
CV (%)5.2%5.3%5.5%6.1%6.4%6.7%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of AGT 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 136434937835336111.243.1%
Sample 223792185239021242269117.065.1%
Sample 373707573834678107774364.724.6%
*number of samples for each test n=16.

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

Gene Name

AGT

Full Name

Angiotensinogen

Weight

53.154kDa

Superfamily

serpin family

Alternative Names

AGT angiotensinogen (serpin peptidase inhibitor, clade A, member 8); angiotensin I; angiotensin II; ANHU; SERPINA8 AGT ANHU, SERPINA8, hFLT1 angiotensinogen angiotensinogen|alpha-1 antiproteinase, antitrypsin|angiotensin I|angiotensin II|fetal-liver predominant transporter 1|pre-angiotensinogen|serine (or cysteine) proteinase inhibitor|serpin A8|serpin peptidase inhibitor, clade A, member 8

*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 AGT, check out the AGT Infographic

AGT 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 AGT: 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].

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8 Customer Q&As for Human AGT/Serpin A8/Angiotensinogen ELISA Kit PicoKine®

Question

Q: what is the protocol regarding preparation of cell lysates prepared for use in Picokine® ELISA kits?

Verified Customer

Verified customer

Asked: 2019-04-25

Answer

A: in those Picokine® ELISAs where cell or tissue lysate is a validated sample type, sample preparation instructions for lysate can be found in the product insert. Components in lysate and lysis buffer can impact immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.

Boster Scientific Support

Answered: 2019-04-25

Question

Q: is it okay to use edta plasma as samples in Human AGT Picokine® ELISA Kit (Catalog # EK1465)?

D. Bhasin

Verified customer

Asked: 2019-04-23

Answer

A: Chelating agents such as EDTA, Heparin and Citrate can attach metal ions from the functional domain of AGT causing degradation of its protein structure. AGT 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 AGT ELISA, treating samples with various anticoagulants and decided that EDTA can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.

Boster Scientific Support

Answered: 2019-04-23

Question

Q: if the enzyme conjugated AGT antibodies are mixed with the substrate, will that convert the substrate into the enzymatic reaction product? Or the enzyme function is only activated when the antibody is attached with the AGT antigen?

Verified Customer

Verified customer

Asked: 2019-02-09

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: 2019-02-09

Question

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

L. Johnson

Verified customer

Asked: 2019-01-10

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 proportion to your positive controls. typically, 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 changeing 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 AGT ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of AGT that can be declared/interpreted as positive by the above standard is 10pg/ml.

Boster Scientific Support

Answered: 2019-01-10

Question

Q: how do I analyze ELISA data? I have obtained AGT level in plasma.

H. Davis

Verified customer

Asked: 2018-11-22

Answer

A: please read this article on ELISA data analysis. bosterbio.com/ELISA-data-analysis-instructions. we also provide a convenient online tool that you can use to analyze ELISA data. bosterbio.com/biology-research-tools/ELISA-data-analysis-online

Boster Scientific Support

Answered: 2018-11-22

Question

Q: which procedure should I follow in order to thaw whole blood sample for AGT ELISA after freezing?

Verified Customer

Verified customer

Asked: 2016-11-17

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 AGT 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: 2016-11-17

Question

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

L. Gonzalez

Verified customer

Asked: 2016-11-15

Answer

A: unable to understand 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 AGT 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: 2016-11-15

Question

Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for measuring AGT using the EK1465 Human AGT Picokine® ELISA Kit?

S. Jackson

Verified customer

Asked: 2014-12-25

Answer

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

Boster Scientific Support

Answered: 2014-12-25

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