Human Serpin D1 ELISA Kit PicoKine®

Serpin D1/Heparin Cofactor II ELISA kit for Human

Human Serpin D1 ELISA Kit PicoKine™ (96 Tests). Quantitate Human SERPIND1 in cell culture supernatants, serum and plasma (EDTA, citrate) and urine. Sensitivity: 0.1ng/ml.

Product Info Summary

SKU: EK1968
Size: 96 wells/kit, with removable strips.
Reactive Species: Human
Application: ELISA
Sample Types: cell culture supernatants, serum and plasma (EDTA, citrate) and urine.

Product Name

Human Serpin D1 ELISA Kit PicoKine®

View all Serpin D1/Heparin Cofactor II ELISA kits

SKU/Catalog Number

EK1968

Size

96 wells/kit, with removable strips.

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

Description

Human Serpin D1 ELISA Kit PicoKine™ (96 Tests). Quantitate Human SERPIND1 in cell culture supernatants, serum and plasma (EDTA, citrate) and urine. Sensitivity: 0.1ng/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 Serpin D1 ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1968)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: NS0; Immunogen sequence: G20-S499

Sensitivity

<0.1 ng/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 EK1968 for more details

Cross-reactivity

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

Reactive Species

EK1968 is reactive to SERPIND1 in Human samples

Validated Sample Types

cell culture supernatants, serum and plasma (EDTA, citrate) and urine.

Application Guarantee

EK1968 is guaranteed for ELISA in Human by Boster Guarantee

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

Background of Serpin D1/Heparin Cofactor II

Heparin cofactor II (HCII), a protein encoded by the SERPIND1 gene, is a coagulation factor that inhibits IIa, and is a cofactor for heparin and dermatan sulfate. It is mapped to 22q11.21. This gene belongs to the serpin gene superfamily. Serpins play roles in many processes including inflammation, blood clotting, and cancer metastasis. Members of this family have highly conserved secondary structures with a reactive center loop that interacts with the protease active site to inhibit protease activity. This gene encodes a plasma serine protease that functions as a thrombin and chymotrypsin inhibitor. The protein is activated by heparin, dermatan sulfate, and glycosaminoglycans. Allelic variations in this gene are associated with heparin cofactor II deficiency.

Kit Components

Catalog Number Description Quantity
EK1968-CAP Anti-Human SERPIND1 Pre-coated 96-well strip microplate 1
EK1968-ST Human SERPIND1 Standard 2 vials, 50 ng/tube
EK1968-DA Human SERPIND1 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.0.0230.0330.3280.4480.8571.0771.4821.954

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect Serpin D1/Heparin Cofactor II, Dilution ratio of 1:10000, concentration in serum and plasma is around 80 ug/ml..

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Human Serpin D1 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)9766436223371236854922654
Standard deviation539.04 302.49 1183.86 70.45 538.59 1449.86
CV (%)4.0%4.7%5.3%5.7%6.3%6.4%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of Serpin D1/Heparin Cofactor II 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 197611609711107105382.187.8%
Sample 264365740587461936060272.054.4%
Sample 32233722710205312110921671885.794.0%
*number of samples for each test n=16.

Gene/Protein Information For SERPIND1 (Source: Uniprot.Org, NCBI)

Gene Name

SERPIND1

Full Name

Heparin cofactor 2

Weight

57.071kDa

Superfamily

serpin family

Alternative Names

HC2; HC2LS2; HCF2; HCF2clade D (heparin cofactor), member 1; HC-II; Heparin Cofactor II; HLS2HCII; leuserpin 2; LS2; Protease inhibitor leuserpin-2; Serpin D1; serpin peptidase inhibitor, clade D (heparin cofactor), member 1 SERPIND1 D22S673, HC2, HCF2, HCII, HLS2, LS2, THPH10 serpin family D member 1 heparin cofactor 2|heparin cofactor II splice isoform|leuserpin 2|protease inhibitor leuserpin-2|serine (or cysteine) proteinase inhibitor, clade D (heparin cofactor), member 1|serpin D1|serpin peptidase inhibitor, clade D (heparin cofactor), member 1

*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 SERPIND1, check out the SERPIND1 Infographic

SERPIND1 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 SERPIND1: 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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9 Customer Q&As for Human Serpin D1 ELISA Kit PicoKine®

Question

Q: Are Boster Bio recombinant proteins and antibodies sterile?

Verified Customer

Verified customer

Asked: 2020-06-08

Answer

A: although the vials are bottled using aseptic techniques, heat-treated vials, and sterile stock solutions, they are not considered or guaranteed to be sterile. If sterile material is required for an experiment, the material can be filtered through a 0.2 micron filter designed for use with biological fluids.

Boster Scientific Support

Answered: 2020-06-08

Question

Q: if the enzyme conjugated Serpin D1 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 Serpin D1 antigen?

V. Dayal

Verified customer

Asked: 2020-05-22

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: 2020-05-22

Question

Q: can you recommend the dilution ratio of serum samples for detection of Serpin D1 in Human serum? I am trying to measure a multiple analytes and it requires 100ul of diluted samples for each well. We have limited sample volumes so we like to dilute as much as possible.

Verified Customer

Verified customer

Asked: 2020-02-14

Answer

A: without having an understanding 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 Serpin D1 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: 2020-02-14

Question

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

M. Jackson

Verified customer

Asked: 2020-02-06

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: 2020-02-06

Question

Q: Can Serpin D1 ELISA Kits be used with tissue homogenates (or other non-validated sample types)?

W. Wright

Verified customer

Asked: 2020-01-23

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 valid for other sample types than we have tested: it means further investigation is needed. 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 assessd 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 view 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: 2020-01-23

Question

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

C. Moore

Verified customer

Asked: 2019-03-15

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: 2019-03-15

Question

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

Verified Customer

Verified customer

Asked: 2018-12-31

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 depends upon the kit used so please refer to each datasheet for details.

Boster Scientific Support

Answered: 2018-12-31

Question

Q: how can I thaw whole blood sample for Serpin D1 ELISA after freezing?

Verified Customer

Verified customer

Asked: 2018-11-30

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 Serpin D1 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: 2018-11-30

Question

Q: What is the optimal O.D. value for Serpin D1 ELISA kit? I performed your Serpin D1 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. can I consider these samples contain Serpin D1 even though the O.D. values are not very high?

Verified Customer

Verified customer

Asked: 2017-11-18

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. an important assessment should be is whether your sample O.D. values are statistically significantly higher than your blank values. considering your assay, 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. 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 Serpin D1 ELISA kit to have sensitivity of 0.1ng/ml, that means the minimum amount of Serpin D1 that can be declared/interpreted as positive by the above standard is 0.1ng/ml.

Boster Scientific Support

Answered: 2017-11-18

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