Product FAQ

What are the differences between TMB and Super AquaBlue?

Answer

TMB is a faster developing, stronger substrate that yields greater amplification/sensitivity/background. The use of an acid stop solution produces a yellow end product read at 450nm. Super AquaBlue is a slower developing blue/green substrate particularly useful for kinetic studies.

Answer Id: E14607

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Product FAQ

What are the components of the Ready-Set-Go! ELISPOT Sets?

Answer

Capture Antibody: Pre-titrated, Functional Grade (low endotoxin) purified antibody
Detection Antibody: Pre-titrated, biotin-conjugated antibody
10X Sterile Azide-Free Coating Buffer
5X ELISA/ELISPOT Diluent
Detection enzyme: pre-titrated Avidin HRP
Certificate of Analysis: Lot-specific instructions for dilution of antibodies and enzyme

Answer Id: E14608

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Product FAQ

Do your Ebioscience™ antibody pairs work in ELISPOT as well?

Answer

Most, but not all, of our ELISA antibody pairs work for ELISPOT. We continue to evaluate and optimize our anti-human and anti-mouse cytokine antibody pairs for ELISPOT applications. Please see our protocols page (www.thermofisher.com/us/en/home/references/protocols/cell-and-tissue-analysis/protocols) for a detailed protocol and a reference table with the list of antibody pairs that can be used in ELISPOT.

Answer Id: E14609

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Product FAQ

I just received a GeneArt™ shipment, but it does not contain all genes from my order. Why is this?

Answer

Partial deliveries can occur when upon reaching the estimated completion date not all order items have been finalized. Partial shipments of incomplete orders are initiated on a weekly basis. Separate shipping of items does not incur an extra cost.
Generally, we ship immediately upon completion of the project item with the longest estimated turnaround time. In rare cases, completely finalized projects are not shipped until the estimated completion date has been reached. Please keep in mind, if you order gene synthesis and subcloning as a process, you will most likely receive the gene synthesis in a Geneart™ vector before you receive your gene in your subcloned vector.

Answer Id: E6686

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Product FAQ

I recently bought one of your Ready-Set-Go! ELISA Sets, can I use the antibody pairs to run an ELISPOT?

Answer

We do not recommend the use of our ELISA kits for the ELISPOT application. We cannot guarantee the successful use of our ELISA kits in an ELISPOT application.

Answer Id: E14610

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Product FAQ

There are two types of MagniSort™ kits – positive and negative selection. When would you choose one over the other?

Answer

The choice between positive and negative selection is best determined by the end users as it is dependent on their specific needs. Cells from both positive and negative selection are viable and can respond to further stimulation.

Answer Id: E14627

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Product FAQ

My array is stuck in the pump on my Applied Biosystems™3500/3500xL Genetic Analyzer, even after loosening the knob. What can I do?

Answer

If the array is stuck in the pump, even after loosening the knob, squirt around the edge of the array head with distilled water and try to remove it again. If the array will not move, a service call may be required. Do not use excessive force or tools to try and remove it.

Answer Id: E12784

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Product FAQ

I’ve previously sent in my own vector to GeneArt™ for an older project. I would like to use this vector again for subcloning my new gene synthesis project. Will I need to resend the vector?

Answer

No, your custom vector has been banked and sequenced and is available to you in your individual short list “My Portal Vectors” as soon as they have been ordered/used once.

Answer Id: E6687

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Product FAQ

How can I reduce background on my PVDF western blots?

Answer

PVDF membranes require more stringent blocking steps. This can be achieved by increasing the concentration of the blocking agent 2-5 fold, increasing the blocking time, and performing the procedure at 37 degrees C. Blocking agents bind to unoccupied sites to prevent background staining and also to membrane-bound proteins, reducing non-specific interactions with the primary antibody. Examples of blocking agents are nonfat dry milk, BSA, and Casein.

Answer Id: E10909

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Product FAQ

What are the critical parameters for successful ELISPOT assay?

Answer

A: It is critically important to use a high-affinity binding PVDF membrane plate, rather than regular ELISA plate or nitrocellulose membrane plate. Additionally, highest affinity antibodies for capture work best in this assay. Empirical optimization of conditions such as cell density, stimulus/mitogen, and kinetics is necessary.

Answer Id: E14611

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Product FAQ

Does MagniSort™ magnetic sorting have an effect on cell viability?

Answer

Cells immediately following magnetic sorting have the same viability compared to unsorted cells.

Answer Id: E14628

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Product FAQ

I need to cut out my SYPRO™ Ruby stained gel bands/spots. What type of light box would be best to use?

Answer

SYPRO™ Ruby dye (as well as SYPRO™ Orange, Coomassie™ Fluor Orange and the nucleic acid stains SYBR™ Safe, SYBR™ Gold, and SYBR™ Green I and II) fluoresces nicely on the Safe Imager™ 2.0 Blue-Light Transilluminator (Cat. No. G6600) or other similar blue light transilluminators with excitation near 470 nm. The gel can be viewed with amber glasses that are supplied with the unit. UV light transilluminators equipped with 302 nm or 365 nm bulbs can be used as well, but would require UV-protective equipment during use.

Answer Id: E11140

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Product FAQ

What is the white residue around the Applied Biosystems™3500/3500xL Genetic Analyzer array knob?

Answer

When placing the array on the instrument, the last step in the process is to loosen the array knob and the instrument forces any air bubbles around the array tip through the threading and the middle of the array knob. Some polymer gets pushed through as well and when it dries up, it appears as a white, flaky residue. Use a damp, lint-free cloth to remove it.

Answer Id: E12785

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Product FAQ

I only have the protein sequence for the gene I’d like to synthesize. Is that okay?

Answer

Yes, we will optimize the protein based on the species you choose to provide you with the optimized gene sequence for your studies.

Answer Id: E6688

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Product FAQ

How should I select a set of primers to use for PCR?

Answer

Here are some general pointers:

Try to choose primer sequences around 50% G + C (plus or minus 15%). If overly G + C rich, add a string of A's or T's at the 5' end. If overly A + T rich, do the same with G's and C's.

Try to avoid G and C at 3' end of the primers. This may increase the chance of forming primer artifacts.

Check primers for self-complementarity, especially at the 3' ends.

To compute an approximate annealing temperature, the following equation can be used to calculate a crude Tm: [4(G + C) + 2(A + T)] - 5 degrees C. Differences of 4-6 degrees C between primers do not seem to affect PCR yield. Ideally, however, the optimal annealing temperature of each primer should match and be within the 55-75 degrees C range.
Use of a computer program may help you design better primer pairs.

Answer Id: E1090

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