Rat Endostatin ELISA Kit PicoKine®

Endostatin ELISA kit for Rat

Rat Endostatin ELISA Kit PicoKine™ (96 Tests). Quantitate Rat Col18a1 in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 15pg/ml.

Product Info Summary

SKU: EK1377
Size: 96 wells/kit, with removable strips.
Reactive Species: Rat
Application: ELISA
Sample Types: cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Product Name

Rat Endostatin ELISA Kit PicoKine®

View all Endostatin ELISA kits

SKU/Catalog Number

EK1377

Size

96 wells/kit, with removable strips.

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

Description

Rat Endostatin ELISA Kit PicoKine™ (96 Tests). Quantitate Rat Col18a1 in cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA). Sensitivity: 15pg/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

Rat Endostatin ELISA Kit PicoKine® (Boster Biological Technology, Pleasanton CA, USA, Catalog # EK1377)

Clonality of Antibodies

See Datasheet for details

Immunogen

Expression system for standard: NS0; Immunogen sequence: H1591-K1774

Sensitivity

<15 pg/ml

Assay Range

156 pg/ml - 10,000 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 EK1377 for more details

Cross-reactivity

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

Reactive Species

EK1377 is reactive to Col18a1 in Rat samples

Validated Sample Types

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

Application Guarantee

EK1377 is guaranteed for ELISA in Rat by Boster Guarantee

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

Background of Endostatin

Endostatin is a naturally-occurring 20-kDa C-terminal fragment derived from type XVIII collagen. It is reported to serve as an anti-angiogenic agent, similar to angiostatin and thrombospondin. And It is produced by proteolytic cleavage of collagen XVIII, a member of the multiplexin family that is characterized by interruptions in the triple helix creating multiple domains, by proteases such as cathepsins. Using a genomic clone as a probe for fluorescence in situ hybridization, Endostatin was mapped the COL18A1 gene to 21q22.3. By immunoprecipitation analysis using membrane fractions of human mammary epithelial cells, It showed that endostatin specifically bound to cell surface nucleolin with high affinity. Blockage of nucleolin with neutralizing antibody or knockdown of nucleolin by RNA interference countered the antiendothelial activity of endostatin and abrogated its antiangiogenic and antitumor activity in vivo.

Kit Components

Catalog Number Description Quantity
EK1377-CAP Anti-Rat Col18a1 Pre-coated 96-well strip microplate 1
EK1377-ST Rat Col18a1 Standard 2 vials, 10 ng/tube
EK1377-DA Rat Col18a1 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)015631262512502500500010000
O.D.0.0320.1750.2940.5110.8511.3201.8252.213

Data Example Images

Recommended Sample Dilution Ratios

According to our internal validation assays using this ELISA kit, to detect Endostatin, Dilution ratio of 1:5, concentration in serum and plasma is around 10 ng/ml..

Intra Assay Consistency & Inter Assay Consistency

We measured random samples of Rat Endostatin 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)2651669431025317364538
Standard deviation18.0295.13219.8121.5112.84231.43
CV (%)6.8%5.7%5.1%%8.5%6.5%5.1%

Reproducibility

We ensure reproducibility by testing three samples with differing concentrations of Endostatin 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 126524927424425812.064.6%
Sample 21669172618591913179198.255.4%
Sample 343104545458138314316299.136.9%
*number of samples for each test n=16.

Gene/Protein Information For Col18a1 (Source: Uniprot.Org, NCBI)

Gene Name

Col18a1

Full Name

Collagen alpha-1(XVIII) chain

Weight

182.172kDa

Superfamily

multiplexin collagen family

Alternative Names

antiangiogenic agent; COL18A1; collagen alpha-1(XVIII) chain; collagen, type XVIII, alpha 1; Endostatin; FLJ27325; FLJ34914; human type XVIII collagen10endostatin; KNO; KNO1MGC74745; Knobloch syndrome, type 1; KS; multi-functional protein MFP Col18a1|endo|collagen, type XVIII, alpha 1|collagen alpha-1(XVIII) chain|alpha-1(XVIII) collagen|endostatin|procollagen, type XVIII, alpha 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 Col18a1, check out the Col18a1 Infographic

Col18a1 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 Col18a1: 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 Rat Endostatin ELISA Kit PicoKine®

Question

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

Verified Customer

Verified customer

Asked: 2019-10-27

Answer

A: we do not recommend freezing 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 Endostatin 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: 2019-10-27

Question

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

Verified Customer

Verified customer

Asked: 2019-10-17

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: 2019-10-17

Question

Q: how are cell lysates prepared for use in Picokine® ELISA kits?

Verified Customer

Verified customer

Asked: 2019-04-13

Answer

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

Boster Scientific Support

Answered: 2019-04-13

Question

Q: Are Boster Bio recombinant proteins and antibodies sterile?

B. Patel

Verified customer

Asked: 2018-12-08

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: 2018-12-08

Question

Q: we need your recommendation regarding the dilution ratio of serum samples for detection of Endostatin in Rat plasma? I am trying to measure a a number of 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.

Verified Customer

Verified customer

Asked: 2018-10-20

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 Endostatin 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: 2018-10-20

Question

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

T. Lopez

Verified customer

Asked: 2018-07-25

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 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: 2018-07-25

Question

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

Verified Customer

Verified customer

Asked: 2018-05-09

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: 2018-05-09

Question

Q: how to proceed with the analysis of ELISA data? I have performed Endostatin level in plasma.

T. Zhao

Verified customer

Asked: 2018-03-13

Answer

A: we recommend you 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: 2018-03-13

Question

Q: if the enzyme conjugated Endostatin 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 Endostatin antigen?

K. Babu

Verified customer

Asked: 2016-06-23

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: 2016-06-23

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