Human Leptin ELISA Kit PicoKine®

Question 1

Question

Dear Technical Support: I am interested with Human Leptin ELISA Kit PicoKine for human population. My query is; how does these Human Leptin ELISA Kit PicoKine work well for urine sample in human adolescent? Is it required to concentrate the sample in order to measure the hormone?

Verified Customer

Verified customer

Asked: 2020-01-27

Boster Scientific Support

Answered: 2020-01-27

Answer

The product Human Leptin ELISA Kit PicoKine has not been tested particularly in human adolescents, however the typical human urine does not need to be concentrated and can be diluted using these Human Leptin ELISA Kit PicoKine if necessary.

Question 2

Question

Q: if the enzyme conjugated Leptin 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 Leptin antigen?

R. Allen

Verified customer

Asked: 2019-12-05

Boster Scientific Support

Answered: 2019-12-05

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.

Question 3

Question

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

Verified Customer

Verified customer

Asked: 2019-10-11

Boster Scientific Support

Answered: 2019-10-11

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 vary slightly by kit so please refer to each datasheet for details.

Question 4

Question

What is the suggested dilution ratio for cell culture supernates using the Human Leptin ELISA Kit PicoKine™ (EK0437)?

Verified customer

Asked: 2019-09-19

Boster Scientific Support

Answered: 2019-09-20

Answer

For cell culture supernates, concentrations of the samples can vary greatly. We suggest to run pilot tests for different group samples to figure out the optimal dilution ratio. Our lab tested unstimulated human peripheral blood mononuclear cells with a dilution of 1:1 and didn't detect any measurable value.

Question 5

Question

What is the suggested dilution ratio for cell lysates using the Human Leptin ELISA Kit PicoKine™ (EK0437)?

Verified customer

Asked: 2019-09-19

Boster Scientific Support

Answered: 2019-09-20

Answer

For cell lysates, concentrations of the samples can vary greatly. We suggest to run pilot tests for different group samples to figure out the optimal dilution ratio. Our lab tested unstimulated human peripheral blood mononuclear cells with a dilution of 1:1 and didn't detect any measurable value.

Question 6

Question

What is the suggested dilution ratio for serum and plasma (heparin, EDTA) using the Human Leptin ELISA Kit PicoKine™ (EK0437)?

Verified customer

Asked: 2019-09-19

Boster Scientific Support

Answered: 2019-09-20

Answer

For serum and plasma (heparin, EDTA), the suggested dilution ratio from normal healthy human is 1:20.

Question 7

Question

I want to purchase your Human Leptin ELISA Kit PicoKine, but I have some questions; does this kit would be useful in rabbit serum?

Verified Customer

Verified customer

Asked: 2019-08-15

Boster Scientific Support

Answered: 2019-08-15

Answer

The product Human Leptin ELISA Kit PicoKine have not been tested for any cross reactivity with rabbit samples, thus we do not know if the kit will work with rabbit serum samples. In this kit both capture and detection antibody are monoclonal so unless the homology between human and rabbit is very high, there would be a possibility that it will cross react, but we cannot guarantee.

Question 8

Question

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

Verified Customer

Verified customer

Asked: 2019-06-20

Boster Scientific Support

Answered: 2019-06-20

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

Question 9

Question

Please help with this. On page 6 of the attached insert...it states the dilution of sample must be made with the sample diluent. But it does not state the target dilutions to be prepared by the user. I looked through this on the attached insert but also cannot understand. Please clarify for me asap Sample Dilution The target protein concentration should be estimated and appropriate sample dilutions should be selected such that the final protein concentration lies near the middle of the linear dynamic range of the assay. It is recommended to prepare 150 ?l of sample for each replicate to be assayed. The samples should be diluted with sample diluent and mixed gently.

Verified Customer

Verified customer

Asked: 2019-06-07

Boster Scientific Support

Answered: 2019-06-07

Answer

Please see the following recommended dilution ratios for Human Leptin PicoKine

Question 10

Question

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

S. Garcia

Verified customer

Asked: 2019-05-29

Boster Scientific Support

Answered: 2019-05-29

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 a must. 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 see 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 Amazon giftcard as an additional reward, if the review is submitted there as well.

Question 11

Question

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

Verified Customer

Verified customer

Asked: 2019-01-14

Boster Scientific Support

Answered: 2019-01-14

Answer

A: for 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 change immunoreactivity, so if lysate is not a validated sample type, care must be taken in sample preparation and validation.

Question 12

Question

Q: how long can samples (cell cultures, serum, and plasma) be stored and still be stable for analysis Leptin using the EK0437 Human Leptin Picokine® ELISA Kit?

D. Varadkar

Verified customer

Asked: 2018-09-19

Boster Scientific Support

Answered: 2018-09-19

Answer

A: Boster Bio does not evaluate sample stability. Variations in sample collection, processing, and storage may change the stabilityof samples. It is recommend to assay sample right after 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.

Question 13

Question

Q: can I use citrate plasma as samples in Human Leptin Picokine® ELISA Kit (Catalog # EK0437)?

Verified Customer

Verified customer

Asked: 2018-04-16

Boster Scientific Support

Answered: 2018-04-16

Answer

A: Chelating agents such as EDTA, Heparin and Citrate can attach metal ions from the functional domain of Leptin causing degradation of its protein structure. Leptin may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes a must for detection and block the antigen antibody reaction. We have tested the Leptin 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.

Question 14

Question

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

Verified Customer

Verified customer

Asked: 2016-06-17

Boster Scientific Support

Answered: 2016-06-17

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 Leptin for a later time, you should let the blood clot in glass tubes and separate the serum to freeze for later analysis.

Question 15

Question

EK0437 Human Leptin PicoKine ELISA Kit I have a customer who is interested in the kit. They want to test it with human serum samples. Please provide your QC data for the current lot of kit along with reference values for serum samples.

Verified Customer

Verified customer

Asked: 2015-05-19

Answer

COA

Boster Scientific Support

Answered: 2015-05-19

Answer

Please see the following recommended dilution ratios for plasma (EDTA) from normal healthy human. leptin (EK0437), 1:20 adiponectin (EK0595) around 1:2000 endothelin (EK0945) 1:1

SKU:	EK0437
Size:	96 wells/kit, with removable strips.
Reactive Species:	Human
Application:	ELISA
Sample Types:	cell culture supernatants, cell lysates, serum and plasma (heparin, EDTA).

Catalog Number	Description	Quantity
EK0437-CAP	Anti-Human LEP Pre-coated 96-well strip microplate	1
EK0437-ST	Human LEP Standard	2 vials, 10 ng/tube
EK0437-DA	Human LEP 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

Human Leptin ELISA Kit PicoKine®

Référence EK0437

Marque : Boster Bio