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Cell Metabolism FAQs

In order to provide better customer support ,Elabscience have compiled and published the most frequently asked questions (FAQs) about cell metabolism assay kit, and will continue to update

  • The activity center of SOD is metal ions, and the use of EDTA as an anticoagulant will chelate metal ions, which may cause the decrease of SOD enzyme activity and affect the results.
  • SDS in RIPA lysis buffer interferes with enzyme activity determination, so it is not recommended to use RIPA lysis buffer for cell treatment. The SOD kit requires that the sample should not contain SDS, Tween20, NP-40, Triton X-100 and other detergents, and should not contain DTT, 2-mercaptoethanol and other reducing reagents.
  • The environment, operation and other influencing factors of each experiment will be a little different. It is recommended to re-examine the control well and control blank well in each experiment in order to obtain more accurate results.
  • It is recommended that cell suspensions that have been homogenized be tested on the same day and not frozen.If there is a temporary accident, you can directly freeze the cells, it is generally recommended that -80℃ not exceed one month.
  • Yes, as long as the temperature can reach 100℃ and the heat is uniform.
  • The control well is mainly set to eliminate the interference of the sample itself. If there is hemolysis or lipemia in the blood sample, it is necessary to set the control well; if there is no such phenomenon, it is not necessary to set the control well.
  • The sample homogenate medium of this kit can only be used with reagent 5 as required in the manual, and cannot be replaced.
  • It is normal that solid precipitation may occur after storage of reagent 3. It is heated and dissolved at 80℃, and can be used normally after cooling.
  • It is a normal phenomenon. After taking it out from -20℃, it can be used after thawing at room temperature.
  • None of the microplate in the metabolism kit can be reused.If the microplate in this kit is not enough, you can use the new black microplate in your own laboratory instead.
  • The kit must be detected by an instrument with chemiluminescence function. Without a chemiluminescence instrument, a multi-function microplate reader with chemiluminescence function can be used, and a ordinary microplate reader cannot be used.When the kit is tested, it only needs to set the chemiluminescence detection, but does not need to set the excitation and emission wavelength.
  • It is recommended to use fresh samples as far as possible. If fresh samples are not available, it is recommended that the samples be frozen at -80℃ for no more than one month, otherwise the AST/ALT enzyme activity will decrease, which will have a great impact on the results.
  • The homogenate medium is the same for both kits. Samples can be processed with the required reagents in either kit, and then tested separately for total iron and ferrous content,but be careful not to mix the other reagents in the two kits.
  • The purpose of the control well setting is to exclude the interference of the sample itself. Each sample requires at least one control well and one sample well.
  • The reagent 1 used in sample preparation will affect the determination of protein concentration, so it can not be calculated by protein concentration, but by cell number.
  • The kit used the matching cell lysis buffer to process the cell samples. After adding the lysis buffer, it only needs to stand on the ice box for 10 minutes, then the supernatant can be centrifuged to be tested, which effectively improves the cell crushing efficiency.
  • The concentration of indicators in the sample is relatively low, and the final color may not be judged by the eye, which is normal, but it can be detected by the instrument.
  • 1 King unit/100 mL = 7.14U/L
  • The phenol red in cell culture medium samples can interfere with results, so this kit is not suitable for cell culture medium samples. For the detection of cell culture medium samples, the E-BC-K766-M kit is recommended.
  • This kit measures the content of (Cu2+) in a sample.
  • If the copper ion content in the sample exceeds the maximum detection value of the kit, the sample needs to be diluted.However, because the copper ion content in cell samples is generally relatively low, this kit does not need to be diluted to detect cell samples.
  • This is a normal phenomenon, and the reagent 1 should be incubated at 37 ℃ until clear before use.
  • It may be that the content of copper ions in the measured cells is low, and is lower than the detection limit of the kit. The number of sample cells can be appropriately increased to increase the concentration of copper ions in the detection sample.
  • It is recommended to use fresh samples as far as possible. If the samples are frozen, the results may be low or undetectable.The longer the frozen storage time, the greater the impact on the results. Generally, the sample should not be stored at -80℃ for more than one month. If the frozen storage time is more than one month, it is not recommended to use.
  • If the tissue weight is less than 0.1g, it can be reduced according to the proportion of tissue weight and homogenate volume as described in the manual. It is necessary to ensure that the homogenated volume of the final sample is sufficient for mitochondrial complex I detection and protein concentration detection.
  • Tissue samples need to be homogenized, and the protein concentration released varies with the degree of homogenization. Therefore, by detecting the protein content in the sample, the difference in the homogenization process of the sample can be corrected to ensure the accuracy of the results. Therefore, tissue samples need to detect the protein concentration.
  • Because the reaction rate of this kit is relatively fast and the control of time is very accurate, it is recommended that the number of sample wells should not exceed 8 at a time to ensure the accuracy of the measurement results. If each sample does 2 double wells, the number of samples tested at one time should not exceed 4.
  • There are differences among different samples. The specific concentration and action time of reagent 1 and the specific concentration and stimulation time of reagent 2 need to be explored by pre-experiment and determined by pre-experiment.The number of samples that can be detected by the kit depends on the concentration and dosage of reagent 1.
  • This kit is suitable for detecting ROS in living cells. After tissue homogenate, cells have broken and died, so this kit cannot detect tissue homogenate.If it is a fresh tissue sample, a single cell suspension is prepared and can be detected.
  • It detects strong green fluorescence. If the sample carry fluorescent green there is interference, other color fluorescence has no effect on the kits.
  • E-BC-F002 kit, laboratory has tested E. coli sample, is suitable.
  • The probe itself is red in the kit and changes from red to green as the lipid peroxide level increases, so you can detect red fluorescence, that is, the flow cytometer selects the cy3 channel instead of the FITC channel.However, it should be noted that the probe concentration should not be too high, and it is recommended that you choose 2-3 samples for pre-experiment.
  • The reagent 4 (Chromogenic Agent A ) of the kit will precipitate in the case of particularly low temperature, which is normal. Before use, incubate at 37℃ for 30 min.
  • The standard is H2S and contains no other proteins. However, there are some proteins in the sample that interfere with the detection, so the sample needs to be treated with reagents 2 and 4, but the standard product is not required.So there are differences in the operation of standard curve and sample determination.
  • Reagent 2 is to dissolve other precipitating substances besides ZnS.
  • Reagent 4 and reagent 5 are acidic, and reagent 2 has been washed in the subsequent operation, so after adding reagent 5, is the reaction system acidic.
  • It depends on the degree of hemolysis.If it's not serious, you don't need to set a control. It doesn't have much effect. If hemolysis is severe, it is not recommended to test, even if the control is set, it may have a greater impact on the result.
  • It can't be replaced with any other reagent, you need to use concentrated hydrochloric acid.
  • The reference dilution of mouse skin tissue was about 20-30 times. The dilution ratio is for reference only. Due to the differences among biological samples, it is recommended to select several samples for pre-experiment to determine the best dilution ratio.
  • In order to eliminate the interference substances of the sample itself and ensure the accuracy of the results.
  • For this kit, tissue samples need to be homogenized with powder A application solution to ensure the MPO extraction effect. If normal saline is homogenized, the measured value may be affected and the result may be low.
  • It may be due to the protein precipitation in the sample, it is recommended to increase the dilution ratio of the sample before testing.
  • There is not much impact, you can set the control well to see the measured value, if the control well OD value is lower than the blank, you can not set the control.
  • This kit detects the content of free calcium in the sample.
  • This kit detects the content of L-glutamic acid in the sample.
  • No, you can't substitute for two acids in different concentrations.
  • Since the sample itself contains creatinine and creatine, the addition of reagent 1 converts the creatine in the sample to hydrogen peroxide. But the creatinine in the sample did not react at the first step, but from the second step; The reaction in the first 2 min is to consume the creatine contained in the sample itself, and this part of the measurement value is deducted to ensure the accuracy of the measurement results.
  • If the purpose of the experiment is to verify the effect of the drug (iron chelating agent), it can be detected with this kit without interference. The probe of this kit is to detect iron ions that have not been chelated, and the probe will not rob the iron that has been chelated.
  • This kit tests for L-glutamine, not L-alanine-glutamine. L-alanine-glutamine was relatively stable in the medium, and the change of L-alanine-glutamine could not be detected by testing the medium samples with this kit.
  • Using the microplate in the kit, it can be installed without spilling.There may be small differences in the maximum volume of microplate wells of different manufacturers, and overflow may occur if other microplate is used.
  • The reference value of mouse serum protein concentration was 50-70g/L and the reference dilution factor was 60-80 times. Due to the differences between biological samples, the above data are for reference only.
  • For samples with severe hemolysis, the effect of setting control is not obvious, it is recommended that you re-sample and use samples without hemolysis for the experiment.
  • It has been documented that the probe can be co-dyed with DAPI
  • The lysis buffer in this kit cannot lyse bacteria.
  • This kit detects the PFK1.
  • The ROS kit does not test for a specific type of ROS, but for total ROS, because the probe is not that specific.
  • There is no need to specially pretreat the sample, and the conversion step is included in the actual detection process.
  • It will affect the enzyme reagents, thus affecting the test results.
  • Yes,it can be used in WB experiment.
  • It can be increased to 40 μL~60 μL.
  • The white microplate can not be used. If the microplate in the kit is used up, the black opaque microplate should be used instead.
  • No, reagents from different kits can not be mixed
  • Milk samples have interference and are not suitable for this kit.
  • Ammonia will be released after the blood sample is isolated, and the value of frozen serum or plasma may be low. It is recommended to use fresh serum or plasma samples for testing.
  • No, can only use normal saline.
  • Saliva samples is applicable
  • No,it can not.
  • It has no effect.