Flow Cytometry Reagents - Faqs
Questions pertaining to BioGems' line of Conjugated Flow Cytometry Antibodies, SAFIRE Functional Antibodies, Purified Monoclonal Antibodies, Flow Cytometry Buffers and Solutions, and Isotype Controls.
What do the application abbreviations stand for?
FC – Flow Cytometry FA - Functional Assay IHC – Immunohistochemistry IF - Immunofluorescence IP- Immunoprecipitation N – Neutralization WB- Western Blot
What does BioGems offer?
BioGems offers a wide comprehensive line of flow cytometry and recombinant protein reagents. We offer a wide range of monoclonal and polyclonal antibodies for the detection of cell surface markers and intracellular cytokines. We sell industry top selling products that are tested to be premium quality at economical prices.
How are quality controls established for different lots?
BioGems offers products that have consistent quality, purity, and performance across multiple lots. New lots are tested side-by-side with previous lots for consistency.
Why are some products listed by number of tests versus µg size?
The products listed by test size are titrated for optimal staining of a cell sample in a final volume of approximately 100 µL. The number of cells in a sample has a typical range of 1x10^5 to 1x10^8 cells. The products listed by µg size are based on a defined concentration as opposed to an optimal staining.
What epitope does the antibody bind to?
BioGems does not at this time perform epitope mapping. In some cases, relying on published research, this information may be listed on the datasheets for certain products
What if I want to use less cells or less volume in my sample that is recommended per test?
As long as the antibody concentration remains the same, you can use less cells or sample volume per test. If you decrease the sample volume to half the recommended, from 100 µL to 50 µL, you can halve the amount of antibody used. If you keep the recommended sample volume, but decrease the concentration of cells, then we recommend you still follow the recommended amount of antibody.
Can antibodies be frozen?
Freezing can denature the product and uncouple fluorochromes. The antibodies should be protected from light and stored at the recommended temperature listed for each product.
What controls should be used with the flow cytometry experiment?
We recommend isotype controls for each flow cytometry experiment and the recommended isotype control can be found on the product page. They retain the non-specific characteristics of the antibody used without the specific target cell binding. Therefore, Isotype controls are important to determine the background staining of an antibody. One can also measure the autofluorescence of a particular cell type with unstained cells.
How much or at what concentration should I use the isotype controls?
The amount or concentration of isotype controls should match used of the primary antibody. Since concentrations between isotype and primary antibodies vary, the same volume of the solutions may not contain equal amounts of antibody.
What is the purity of the antibodies?
The monoclonal antibodies are >90% pure as determined by SDS-PAGE.
What is the purpose of Tandem Dyes?
Tandem dyes are composed of two covalently attached fluorescent molecules. Through the fluorescence resonance energy transfer (FRET) principle, the tandem dye utilizes the excitation spectrum of the donor fluorochrome and the emission property of the acceptor molecule. This property allows for larger stokes shifts, the difference between the excitation and emission range.
What is Compensation and why is it important?
Fluochromes do not emit at distinct wavelengths but across emission spectrums. Compensation is the process of removing the overlapping spectral regions to eliminate false signals in multicolor staining panels.
For products with multiple clones, how should one choose between them?
We recommend basing your decision on a search on Google Scholar or Pubmed. These resources can help you research the popularity of these clones compared to each other for your relevant purposes.
Small Molecules - Faqs
Questions pertaining to BioGems' line of Small Molecules
What are Small Molecules?
Small Molecules are bioactive low molecular weight organic compounds that can be used in a diverse number of life science research applications. Their small size allows for the possibility of rapidly diffusing across cell membranes to reach intracellular sites of action. They are high purity chemical defined reagents with exceptional lot to lot consistency.
What is the product purity and how is it determined?
Our small molecules are of very high purity, typically >98%. The purity and quality of each lot is tested using a comprehensive array of techniques, including NMR, optical rotation, HPLC, TLC, and mass spectrometry.
What is the CAS number?
The CAS number listed on our datasheets is the unique number assigned to every chemical described in literature by the Chemical Abstracts Service of the American Chemical Society.
How should I dissolve the small molecule that I purchased from you?
The product datasheet includes information on the compound’s solubility. Many compounds that are not directly soluble in water can be dissolved in alcohols or solvents, such as ethanol or DMSO, to create a concentrated stock solution. These stock solutions are often reasonably stable for storage, and can be diluted to the desired final concentration with water or aqueous media at the time of use.
What is the highest DMSO concentration to use in cell culture?
Typically, concentration less than 0.5-1% DMSO is tolerable by many cancer cells in culture. For more sensitive cells such as primary cells, it is recommended to use less than 0.1% DMSO in the cell culture.
The small molecule does not fully dissolve at the desired concentration. What should I do?
The easiest way to speed up solubilization is often to warm the solution. In addition, we recommend vortexing or sonication for several minutes to dissolve the compound.
The products TDS sheet states that the product should be stored at -20°C, but the product arrived at room temperature. Can I still use this product?
Many small molecules are validated to be stable at warmer temperatures for short periods of time and can be shipped at ambient temperatures. Please follow the recommended storage instructions on the label or product datasheet once you receive the products for optimal long term storage.
What is the amount of small molecule I should use for my experiment?
We recommend that you review published literature for guidelines on how to best use our products for your specific application. We do not carry out any biological studies of our products in vivo.
ELISA KITS - Faqs
Questions pertaining to BioGems' line of Pre-Coated ELISA KITS.
What is the advantage of using Pre-Coated ELISA kits?
BioGems Pre-Coated ELISA kits are designed to be complete, ready to go, and easy to use. They save time by eliminating the need to pre-coat the ELISA plates.
Can I use different components from different companies for my ELISA?
Each of the BioGems Pre-Coated ELISA kits is developed, validated, and optimized using the reagent concentrations and protocols that are included in the kit. Any changes to the reagents or protocols can negatively affect the final assay performance.
I ran out of some ELISA kit components, can I buy separately?
Yes, please contact us directly at firstname.lastname@example.org for the specific details regarding any component in the kit.
Can a partial ELISA plate be used?
Yes, the ELISA plates have removable well strips. The unused wells maybe be removed form the plate, returned to the foil pouch and stored at 2-8° C for later use.
What could be causing high variability between sample duplicates?
The most likely reasons for high variability in an assay are due to washing and pipetting issues or introducing variations to the protocol.
Why doesn't the assay range extend to the stated sensitivity?
Sensitivity is the lowest measurable value that is statistically not equal to zero and it is calculated based on the signal of the background and the inherent variability of the assay.
What is the well depth of the ELISA kits?
The well depth is 300 ul and the max capacity of each well is 350 ul.
What is your validation process?
Every kit we offer is subjected to an array of rigorous product-specific quality controls before delivery. They are evaluated for intra-assay and inter-assay reproducibility and all testing results are compared to previous lots for lot-to-lot consistency.