Rat MMP-2 ELISA Kit PicoKine®

Question 1

Question

Q: for how much duration can samples (cell cultures, serum, and plasma) be stored and still be stable for quantification MMP-2 using the EK0639 Rat MMP-2 Picokine® ELISA Kit?

G. Anderson

Verified customer

Asked: 2018-12-14

Boster Scientific Support

Answered: 2018-12-14

Answer

A: Boster Bio does not assess sample stability. Variations in sample collection, processing, and storage may alter the stabilityof samples. It is recommend to assay sample immediately after collection when possible, or aliquot into single use volumes and store samples frozen. avoid repeat freeze-thaw cycles with the stored samples in order to prevent protein degradation.

Question 2

Question

Q: which procedure should I follow in order to thaw whole blood sample for MMP-2 ELISA after freezing?

W. Tiwari

Verified customer

Asked: 2018-12-11

Boster Scientific Support

Answered: 2018-12-11

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 MMP-2 for a later time, you should let the blood clot in glass tubes and collect the serum to freeze for later use.

Question 3

Question

Q: can you recommend the dilution ratio of serum samples for detection of MMP-2 in Rat 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 volumes so we like to dilute as much as possible.

Verified Customer

Verified customer

Asked: 2018-07-01

Boster Scientific Support

Answered: 2018-07-01

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 MMP-2 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.

Question 4

Question

Q: Can MMP-2 ELISA Kits be used with tissue homogenates (or other non-validated sample types)?

S. Banerjee

Verified customer

Asked: 2018-05-17

Boster Scientific Support

Answered: 2018-05-17

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

Question 5

Question

Q: What is the optimal O.D. value for MMP-2 ELISA kit? I used your MMP-2 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 MMP-2 even though the O.D. values are not very high?

M. Johnson

Verified customer

Asked: 2018-04-19

Boster Scientific Support

Answered: 2018-04-19

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 the above 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 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 MMP-2 ELISA kit to have sensitivity of 10pg/ml, that means the minimum amount of MMP-2 that can be declared/interpreted as positive by the above standard is 10pg/ml.

Question 6

Question

Q: if the enzyme conjugated MMP-2 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 MMP-2 antigen?

Verified Customer

Verified customer

Asked: 2016-11-14

Boster Scientific Support

Answered: 2016-11-14

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 7

Question

Does EK0639 detect active MMP-2 or the full MMP-2 rat protein? Will it cross-react with other proteins?

Verified Customer

Verified customer

Asked: 2016-09-12

Boster Scientific Support

Answered: 2016-09-12

Answer

EK0639 is able to detect the full MMP-2 rat protein including zymogen and active MMP-2. It cross reacts with recombinant human MMP-2. Recombinant mouse MMP-3 protein, mouse MMP-12 protein and rat MMP-9 protein was also tested and no significant cross reactivity was observed.

Question 8

Question

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

Verified Customer

Verified customer

Asked: 2015-09-05

Boster Scientific Support

Answered: 2015-09-05

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.

Accepted Answer

A: Chelating agents such as EDTA, Heparin and Citrate can bind metal ions from the functional domain of MMP-2 causing degradation of its protein structure. MMP-2 may be denatured as a result and may compromise the assay's measurements. The chilating sites could also be too close to the epitopes needed for detection and block the antigen antibody reaction. We have tested the MMP-2 ELISA, treating samples with a number of anticoagulants and decided that heparin can be used for treatment of blood/plasma samples. Do not use other anticoagulents when collecting samples.

SKU:	EK0639
Size:	96 wells/kit, with removable strips.
Reactive Species:	Rat
Application:	ELISA
Sample Types:	cell culture supernatants, serum and plasma (heparin).

Catalog Number	Description	Quantity
EK0639-CAP	Anti-Rat Mmp2 Pre-coated 96-well strip microplate	1
EK0639-ST	Rat Mmp2 Standard	2 vials, 40 ng/tube
EK0639-DA	Rat Mmp2 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

Rat MMP-2 ELISA Kit PicoKine®

Cat# EK0639

Brand : Boster Bio