Starch Assay Kit (ab83393)

Thank you for your response.

My colleague is out of office for a couple of days this week and she has asked me to look after her customers during her absence.

I am very sorry that your customer is having a hard time extracting starch from the seeds. Since we have not used this kit with seeds in-house, I can only make some suggestions as to what may help.



1) Longer incubation (more than 5 min) on the heating block might help.

2) Use more seeds initially (like 20 mg) to start seeing the pink colour development.


3) Increase the time of incubation in step 5, as I suggested in my previous email.



I can't comment on the salt concentration as that might be seed specific issue.



Hope this information has been helpful for you. Please let me know if you have any other questions.



Thanks,

Thank you very much for that feedback. This sort of information is indeed very important for your customers.

I would strongly encourage your customer to provide us with an Abreview for the product. This would simply involve sharing the protocol details of the experiment and some data obtained which I would then add to the datasheet of the product.

More information on the Abreview system can be found here:

https://www.abcam.com/Abreview

As a reward your customer would receive Abpoints which can go towards a future Abcam purchase, or can be claimed as Amazon vouchers. Information on the Abpoints can be found from the following link:

https://www.abcam.com/abpoints

I hope this information has been of help. If your customer would like to submit the Abreview they will need to do it directly to me as we are currently unable to support Abreviews for kits online. We are hoping to remedy this soon.

Please do pass on our thanks to the customer and I hope they will consider submitting an Abreview.

Have a nice weekend.

Thank you once again for your enquiries which have been forwarded to me as my colleague Karin is currently away from the office.

Karin has discussed this withour source for this antibody, and they have kindly provided the following advise:

1. All the details described in the protocol on the datasheet are standardized for samples different from these seeds. In order to get comparable results with these seeds, some tweaking of the details/optimization has to be performed by the end user. So the 30 min incubation time after which the absorbance is still increasing, might indicate that this time has to be standardized for these samples. I would suggest a time course experiment with times ranging upwards of 30 min, to see the optimal end point time.

What was the maximum time at which the OD was measured?

2. I agree the standard readings are very low. It may be due to various reasons:

- Improperly thawed components - Thaw all components completely and mix gently before use

- Use of expired kit or improperly stored reagents - Always check the expiry date and store the components appropriately

- Allowing the reagents to sit for extended times on ice - Always thaw and prepare fresh reaction mix before use

- Incorrect incubation times or temperatures - Refer datasheet & verify correct incubation times and temperatures

- Incorrect volumes used.

It might also happen if the OD is not measured at exactly 570 nm.

I hope this information will be helpful and help optimize the results from the sample. However, I am concerned that. Please let me know how the customer gets on and how they would like to proceed in this case.

Thank you for contacting us.

I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

In order to understand more fully what the problem may be could I please ask you to answer the following questions:

1. Have you performed the standard curve? Has this produced the expected graph?

2. Could you describe exactly how the samples were obtained, extracted and washed?

3. Was a glucose control performed (not using hydrolysis enzyme)? What were the result of this reading?

4. You say that you see erratic readings "it changes a lot every few minutes", does it just go up, or up and down? What values are you seeing?

5. Are the values seen within the expected detection limit of the kit (0.0004-2 mg/mL)?

6. Were the buffers used allowed to come to room temperature prior to carrying out the protocol?

With this information I will hopefully understand more fully what may be contributing to the problems you have been observing.

I look forward to receiving your reply.

Thank you for contacting us.

Theoretically this kit can be used with any type of starch containing samples. I assume it can detect Starch inleaves of Arabidopsis thaliana also. Please be advised that we haven't used this kit for detection of starch inleaves of Arabidopsis thalianaso we will not be able to provide any tested data. The optimal experimental conditions has to be determined by the end user.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Thank you for contacting us.

Theoretically this kit can be used with any type of starch containing samples. I assume it can detect Starch in legume seeds also. please be advised that we haven't used this kit for detection of starch in legume seeds so we will not be able to provide any tested data. The optimal experimental conditions has to be determined by the end user.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.