深圳欣博盛生物科技有限公司

1. Why does the vial I received appear empty?

Lyophilized powder may rest in the bottom angle of the vial or near the   stopper. Add an appropriate solvent as indicated on the datasheet to dissolve   the product for use.
  Note that some of our products do appear as clear/ translucent films which   can be harder for visual inspection, e.g. MPLA, CRX-527, PMA and LyoVec™.

2. My product has arrived damaged. What should I do?

Please report to us with photos of the damage and specify the PO and LOT   number.

3. Why is catalogue code printed on the vial label sometimes   different from the official one?

As some of our catalogue codes had been updated, the old version of cat   code may remain on the vial label.
  E.g. The vial label of vac-pova shows vac-efova instead. vac-efova was the   old product code of vac-pova.
  In case of some products, catalogue codes on vial label are our internal   production reference, they may differ from the one on offical label.
  E.g. vial label: tlrl-nacgas, product label: tlrl-nacga

1. What   can I do if my product is not dissolved at the instructed concentration and   solvent?

In general,   customers can warm the solution at 37°C and vortex until complete   resuspension. If it still does not re-dissolve,  please contact our   technical support for further assistance.
  Note that it can be difficult to solubilize lipids like MPLA by vortex. The   suspension may appear to contain floating fine particles. Further sonication   (e.g. at 25KHz 35°C for 5 mins) can help disperse these particles.

2. Can   I use the solvent other than suggested by TDS?

We recommend   customers to use our instructed solvent to obtain optimal dissolution.
  E.g. Conventional R848 is poorly soluble in water and soluble in organic   solvents. Yet, InvivoGen's R848 is designed as water soluble, customers can   dissolve it using the endotoxin-free water provided.
  E.g. Some may use isopropanol to solubilize TDB, yet our internal testing   found that it does not dissolve well in isopropanol at 1mg/ml. We recommend   customers using DMSO as suggested by TDS .

3. Can   I prepare a higher stock concentration than instructed?

We recommend   customers to prepare the stock solution at instructed concentration.
  E.g. Poly (I:C) already forms a thick mixture when dissolving at 1mg/ml.   Though heating at 65-70°C for 10mins would mix well, we do not advise a   higher stock concentration than 1mg/ml, as it will be more difficult to   solubilize Poly (I:C) and may affect the annealing process.

1. How   long will the product be stable? Where can I find the expiry date of the   product?

The   stability information is stated in TDS. If handled and stored appropriately,   the product should be stable up to the expiry date assigned. While the   expiry/retest date of the product is indicated on Certificate of Analysis,   you can request the CoA from our customer service, specifying the catalogue   code, LOT number (and PO number if available).  
  Note that for antibody and plasmid products, there are no expiry dates/retest   dates indicated.

2. Can   the product be stored at a different temperature? 

The storage   temperatures that we indicate on TDS are the optimal temperature for storage.   We do not guarantee the stability of the product being stored at a different   temperature.

3. If I   cannot finish using the product at once, can I store the remaining product?

Repeated   freeze/thaw the remainings will affect product activity. We recommend   preparing the aliquots in advance.

4. Why   does the stability of LyoVec™ - complexed ligand products last much shorter   after resuspension?

Products   pre-complexed with transfection reagent will have a shorter shelf life   compared to naked products. Upon resuspension, the pre-complexed product is   only stable for 1 week at 4 °C. If you would like to extend the stability,   please purchase the naked product and transfection reagent separately. Upon   resuspension, the naked product is stable for 1 month at 4°C and 1 year at   -20°C.

1. What   is the guideline to bulk discount?

There is no   fixed order quantity to get a bulk discount. Please contact us to request a   quotation for bulk ordering.

1. Why   is the date of signature in CoA later than expiry date sometimes?

This may   happen on a re-issued CoA for products with expired retest dates. The date of   signature is updated whenever our QA/QC department edits the CoA.

2. Do   you have GMP certificates for production?

No. We do   not offer any GMP certified product. All of our products are strictly for   research use only and not for human or veterinary use.

3. Do   you have toxicological information about the product?

Toxicological   data is not available as stated in MSDS Section 11 (MSDS can be found on each   product website).

4. Where   can I find sterility information?

You can   refer to TDS or CoA which states the absence of bacterial contamination (e.g.   lipoproteins and endotoxins) in our products.

1. Do you have any half life data of your products?

No, we do not perform half-life testing.

2. Do you have in vivo data for ligands products? Can I use them in   vivo?

We do not have in vivo testing in house. We suggest performing   literature search as a reference first.

3. Why can't I find the molecular weight for some of your products?

It is possible that the molecular weight of some products cannot be   determined accurately due to their intrinsic structure or insolubility.
  E.g. PGN is a vast polymer consisting of numerous monomers.  We do   not know how many monomers make up our purified PGN.
  E.g. The nature of LPS causes it to form aggregates of unpredictable sizes   (an estimated M.W. could be about 50 kDa).

4. What is the difference between Poly (I:C) HMW and Poly (I:C)   LMW? Any tips to choose between them?

The primary difference is their molecular   weight. Poly(I:C) HMW is 1.5-8 kb while Poly(I:C) LMW is 0.2-1 kb in size.   Also, the efficiency of TLR3 activation by Poly(I:C) HMW was significantly   higher than that by Poly(I:C) LMW, as tested by our 293/TLR3 and Ramos-Blue   cells. For details visit   https://www.invivogen.com/sites/default/files/invivogen/old/docs/Poly_I_C_LMW_v4-invivogen.pdf 
  Poly (I:C) HMW is therefore recommended if customers are targeting TLR3   activation; for customers studying RIG-I pathway, they can consider choosing   Poly (I:C) LMW.

5. Do you have the exact molecular weight for Poly (I:C)?

The molecular weight of Poly (I:C) (also Poly(dA:dT) and Poly(dG:dC))   can vary from batch to batch. The lot-specific molecular weight is available   to check upon request.

6. What is the advantage of your Poly(I:C) over Sigma's?

A study has shown that InvivoGen poly(I:C) exhibited   negligible endotoxin level and lower inter-batch variability over Sigma   Poly(I:C). For details visit https://doi.org/10.1016/j.bbi.2019.08.006

7. Why is it necessary to transfect Poly (I:C) for activating   RIG-I/MDA5 but not for TLR3?

Naked Poly (I:C) can be internalized into cells by   endocytosis to activate the endosomal TLR3; RIG-I and MDA5 are cytosolic   sensors, Poly (I:C) being transfected can reach the cytosol for activation.

8. I can’t find the purity percentage value of LPS products on the   website. Do Ultrapure products have a higher purity than Standard products?

The two versions of our LPS differ in terms of functional purity.   Standard version stimulates both TLR2 and TLR4 as it contains contaminating   lipoproteins that non-specifically activate TLR2. Ultrapure stimulates only   TLR4, having successive steps of enzymatic hydrolysis to ensure no TLR2   activity.

9. Do you provide the endotoxin concentration for LPS products?

Ultrapure LPS product has the specific endotoxin level (EU/mg) measured   for each lot by LAL assay (the data can be found on CoA).  
  For Standard LPS products, one EU approximately equals one ng of endotoxin,   e.g. 5 mg/ml of Standard LPS-EB solution corresponds to 5 x 10⁶ EU/ml.

10. What is the difference between your LPS products? Any tips for   choosing the LPS?

LPS products differ from species to species, primarily in the O-antigen   hydrophilic sugars' length, sequence and self-linkages.
  LPS may work differently on cell types based on their strains. As a   reference, the most described LPS in the literature is from O111:B4, LPS-EB   is therefore a popular choice.

11. What is the difference between CpG-A, CpG-B and CpG-C?

They are different based on their structural characteristics and   activity on human peripheral blood mononuclear cells.
  CpG-A ODNs induce high IFN- α production from pDC but are weak stimulators of   NF-kB and pro-inflammatory cytokine production.
  CpG-B ODNs stimulate strong B cells and NF-kB activation but are weak   activators of type I IFN secretion.
  CpG-C ODNs combine properties of both class A and C, strongly stimulating B   cells and type I IFN secretion.
  For details visit https://www.invivogen.com/cpg-odns-classes

12.  Is it necessary to transfect flagellin for NLRC4   inflammasome activation?

We recommend transfecting flagellin for NLRC4 activation, however, this   is cell-type dependent. If the cells stably overexpress NLRC4 (e.g. our   THP1-NLRC4 Cells), it is possible that flagellin without the transfection   agent can activate NLRC4, otherwise, it will require a very high   concentration of flagellin to be sufficiently delivered into cytosol for   stimulation.

13.  Does flagellin-induced NLRC4 activation require LPS   priming?

It is common to have LPS priming before flagellin stimulation. LPS is   used for pro-IL-1β, pro-Caspase-1 and NLRP3 induction, and these proteins   will help in the NLRC4 inflammasome activation.

14.  Is transfection of cGAMP needed for stimulation?

InvivoGen does   not transfect the on-shelf cGAMP for stimulating the reporter cell lines.   However, as cGAMP has a poor membrane permeability. You can use a   lipid-based transfection reagent to facilitate cGAMP's delivery to cells for   stimulation.

15. Do you provide IC50 and/or EC50 value for the product?

You may find the values on TDS or validation datasheet for some of our   products.

16. Where can I find the sequence of primer that your plasmids   used?

The information is available on our website:   https://www.invivogen.com/sequencing-primers

17. The pVITRO1-MCS plasmids carry two elongation factor 1 alpha   (EF-1α) promoters, from rat and mouse origins combined to the CMV and SV40   enhancers respectively. Will both promoters display activity in human cells?   And will the rat EF1 promoter display activity in mouse cells?

As EF-1α promoters can be expressed in mammalian cells, they will   display activity in human cells, and the rat EF1 promoter will also display   activity in mouse cells.

18. Can a human promoter work in mice?

Our composite promoters allow for strong expression in nearly any   mammalian cell line. In fact, matching the species of origin for the promoter   to the organism it is being used in (in vitro or in vivo) does not seem to   have much of an impact on gene expression levels. The activity of a promoter   may vary between cell types, but in general a mammalian promoter should work   in any other mammalian model.  In our experience promoters from   mammalian origin tend to work similarly between human and mouse   cells.  A good example is for the IFN-beta promoter used in the   majority of our HEK-Blue-TLR reporter cells. This is the mouse IFN-beta   promoter, however the stimulation of SEAP following TLR induction works   perfectly well in the human HEK293 cell line.

19. Do you have a dual promoter vector with fusion proteins tags   for each?

We do not have a dual promoter plasmid with two fusions, but a strategy   would be to subclone from our other tagged plasmids. For example while our   pVIVO allows for dual expression, it does not allow for GFP fusion. Further,   while pDUO allows for dual expression, it does not include GFP at all (either   as a fusion or marker).
  Only our pSELECT-Tag plasmid allows for fusion to GFP. That plasmid only   allows for insertion of one gene. A strategy to express two genes with 1   fused to GFP would be to first clone onto pSELECT-Tag and then subclone onto   pVITRO or a similar dual expression plasmid.

20. How can I avoid light interference when plating QUANTI-Luc™?

QUANTI-Luc™ is light-sensitive. The light interference can result in   great variation within a single plate. Recommended steps we take to avoid   light interference: wrap QLC in foil/paper, turn off hood lights, work fast,   cover plate while walking to reader.

21. What is the difference between QUANTI-Blue™ and QUANTI-Luc™?

Both are secretory so there is no need to lyse cells, and the choice   really depends on your research need.

You may refer to the following information for selection:
                                                QUANTI-Blue™                  QUANTI-Luc™
  Detection:                            630-655nm OD         vs         Luminescence
  Detection apparatus:         Spectrophotomer      vs         Luminometer
  Sensitivity:                           Lower                        vs         Higher
  Signal stability:                    Higher                       vs          Lower

22. Can QUANTI-Luc™ be used to detect Firefly Luciferase? Can it   detect luciferases dually?

QUANTI-Luc™  does not detect Firefly Luciferase. It detects   Lucia and Renilla Luciferase.