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Click the image above to start asking questions! 

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Q: I have lysed sample using RIPA buffer and store the sample in -80C freezer immediately before running multiplex. How long the sample can be stored in snap freeze? 

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Q: I have lysed sample using RIPA buffer and store the sample in -80C freezer immediately before running multiplex. How long the sample can be stored in snap freeze? 

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A: Proteins can be stored at -80°C for an indefinite period, but the key factors are the method of freezing and the number of thawing cycles. It's advisable to store protein samples as aliquot to minimize freeze-thaw cycles. Properly frozen proteins can remain stable for many years.

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Q: There are kits with DAB and without DAB, what is the difference? 

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Q: There are kits with DAB and without DAB, what is the difference? 

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A: 2-step plus detection kit is a two-step immunohistochemical broad spectrum detection reagent. It polymerizes monovalent Fab fragments of secondary antibody and enzyme, which replaces the secondary antibody and tertiary antibody in traditional method, can directly amplify the binding signal of antibody-antigen. Compared with the traditional SP three-step method that use DAB, this kit has the characteristics of simple, rapid, high-sensitivity. It also involved lesser steps to work with.

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Check out the service PRIMEPLEX provides for flow cytometry!

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Check out the service PRIMEPLEX provides for flow cytometry!

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1. Apoptosis

2. Cell cycle analysis

3. Mesenchymal stem cell (MSc) characterisation

4. Other immunophenotyping aside from MSc

5. Lymphocyte proliferation

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Check out the service PRIMEPLEX provides for cellular research!

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Check out the service PRIMEPLEX provides for cellular research!

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1. Cell culture

2. COMET assay

3. MTT assay

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Check out the service PRIMEPLEX provides for animal research:

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Check out the service PRIMEPLEX provides for animal research:

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1. Project consultation

2. Toxicology & safety assessment

3. Haematology

4. Histology processing & staining

5. Histology report analysis

6. Disease model development

7. Research model advisory

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Q: I have extracted tissue samples and yet to lyse it to produce homogenate. Should I proceed with lysis process and store it or otherwise?

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Q: I have extracted tissue samples and yet to lyse it to produce homogenate. Should I proceed with lysis process and store it or otherwise?

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A: Once sample is lysed, protease is released from the organelle. For long term storage, if the lysis buffer is supplemented with protease inhibitor, they shouldn’t differ much. However, we would recommend to store sample as intact in -80C freezer if protease inhibitor is out of reach.

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Q: Can we use tissue treated with RNAlater and histology block to run ELISA? I want to do ELISA but half of the samples already treated with RNAlater another half in tissue block

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Q: Can we use tissue treated with RNAlater and histology block to run ELISA? I want to do ELISA but half of the samples already treated with RNAlater another half in tissue block

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A: Once the sample is in paraffin wax, the protein and RNA has been disrupted. A paraffinated sample or tissue block is not suitable for ELISA. However, RNAlater sample is feasible for ELISA .

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Q: I am targeting cytokine in my study. How long can the sample with cytokine be kept in -80C?  

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Q: I am targeting cytokine in my study. How long can the sample with cytokine be kept in -80C?  

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A: you can keep the sample for 2 years frozen with approx 5% degradation. Storage above 5 years might result in significant lost of cytokine level in the sample.

reference: Keutermans et al. (2013) Cytokine assays: An assessment of the preparation and treatment of blood and tissue samples. Method

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Q: if I didn’t use sample diluent with my sample, will it affect ELISA kit performance? 

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Q: if I didn’t use sample diluent with my sample, will it affect ELISA kit performance? 

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A: Antibody binding happens in a controlled buffer environment i.e. correct pH and salt content. Hence, it is recommended to use sample diluent to dilute the sample prior to running.

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Q: I intend to use human tears sample on ELISA. Do you think Elabscience ELISA kit can perform on tear sample? 

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Q: I intend to use human tears sample on ELISA. Do you think Elabscience ELISA kit can perform on tear sample? 

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A: All biological fluid is compatible with Elabscience ELISA kit, including tears. A good start when comes to sample other than serum /plasma would be to perform a literature search on the concentration range of this protein in tears to understand the amount of tears required to perform this test. Apart from that, we recommend performing a serial dilution on your sample in a pilot test prior to consuming the whole kit.

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Q: If I use RIPA buffer to extract protein, do I need to use protein inhibitor as well?

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Q: If I use RIPA buffer to extract protein, do I need to use protein inhibitor as well?

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A: It is a good practice to use protease inhibitor cocktail when comes to protein extraction. Generally it is advisable to put in protease inhibitor, especially when it comes to ELISA. Unless you are working with sample that you are going to thaw once or twice only, you might experience loss of protein of interest if there is no protease inhibitor with multiple free-thaw cycle.

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Q: I need PBS and protease inhibitors to homogenize tissue samples before proceeding to elisa. Do you know any products that provide both together or need to be purchased separately? 

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Q: I need PBS and protease inhibitors to homogenize tissue samples before proceeding to elisa. Do you know any products that provide both together or need to be purchased separately? 

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A: Typically most RiPA buffers are supplied without protease and phosphatase inhibitor. However, some brands offer them as a kit, we will recommend checking the datasheet of your product to understand more. Alternatively, you can consider RIPA Lysis Buffer which comes with protease and phosphatase inhibitors.

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Q: I wonder if I can send in my sample before my study is completed? My samples will come in 2 batches at different timing. 

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Q: I wonder if I can send in my sample before my study is completed? My samples will come in 2 batches at different timing. 

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A: We recommend all replicate i.e. technical or biological replicate to be analyzed in the same plate so that they are derived from the same standard curve. This approach is to avoid batch-to-batch variation

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Q: For samples that have been put in thiazole solution, can it be used for elisa afterwards? 

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Q: For samples that have been put in thiazole solution, can it be used for elisa afterwards? 

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A: Depending on the specificity of the antibody, it might work, it might not work. Trizol due to its composition of chloroform and heavy salts will denature the protein/antigen to be detected in ELISA. We do not recommend to run ELISA using sample extracted with Triazole solution.

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Q: I have a question on protein normalisation. I have done bradford assay untuk determine total protein of each samples.Then, after I got my reading from ELISA,.I normalise the result from ELISA by dividing the concentration got from ELISA with total protein got from bradford. Is this acceptable? 

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Q: I have a question on protein normalisation. I have done bradford assay untuk determine total protein of each samples.Then, after I got my reading from ELISA,.I normalise the result from ELISA by dividing the concentration got from ELISA with total protein got from bradford. Is this acceptable? 

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A: Commonly, normalisation of sample with total protein is done prior to ELISA procedure to ensure that all sample started with the same amount of protein. It is also used to ensure that the sample concentration falls onto the detection range of the ELISA Kit. If you can detect the protein of interest with your current practice and do the normalisation after the ELISA procedure , this practice is acceptable.

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Q: For ELISA what is the storage temperature if I already open it once? I run twice for a kit. So I want to keep for the next round at -20. Is this temperature enough?

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Q: For ELISA what is the storage temperature if I already open it once? I run twice for a kit. So I want to keep for the next round at -20. Is this temperature enough?

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A: Not all component in an ELISA kit can be stored in -20C. Protocol manual will suggest the suitable storage temperature for each components or the entire kit. We recommend adhering to the storage temperature advice by the kit manual.

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Q: In ELISA kit protocol, it mentions need to incubate the plate for 1 hour at 37C. My lab does not have incubator for ELISA plate, only for cell culture. Can I ask if I can use water bath for incubation?

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Q: In ELISA kit protocol, it mentions need to incubate the plate for 1 hour at 37C. My lab does not have incubator for ELISA plate, only for cell culture. Can I ask if I can use water bath for incubation?

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A: A CO₂ incubator is a common equipment in cell culture lab. if client are working in a cell culture lab, she can use a CO₂ incubator. To avoid contamination, we do not recommend water bath as the plate is not suppose to be in contact with water inside water bath. We recommend setup laboratory equipments prior to the initiation of test.

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Q: For ELISA, can I reduce the volume of samples in each well into half from suggested in protocol? Because the volume for each of my sample is not up until 100uL. If yes, should I reduce all other reagents into half too including standards and control?

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Q: For ELISA, can I reduce the volume of samples in each well into half from suggested in protocol? Because the volume for each of my sample is not up until 100uL. If yes, should I reduce all other reagents into half too including standards and control?

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A: It is recommended to assess the sample volume prior to consuming the kit. We suggest adhering to the standard and control volume as recommended by the kit. As per the sample volume, since the condition of assay is altered, a pilot test is recommended. The pilot test can include a pre-diluted sample and an undiluted sample with reduced volume to gauge if the modified protocol will archive acceptable result.

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What method Should I go for?IHC or IF to detect M1 and M2 macrophages along with their cytokine receptors and nutrient transport proteins in the same tissue sample? Or need to see the markers in different tissue slides would be beneficial?