HMGB1 AAV (Rat) (CMV) (GFP) (AAV Serotype 1)

CAT.NOUNITPRICE
AAVP69467105 x 200μl
$0.00

Specifications


DescriptionThis ready-to-use AAV is part of abm's AAV Expression System and can be used directly to transiently over-express your gene of interest in a wide range of host cells or animal models.
SKUAAVP6946710
Accession NumberNM_012963
Unit quantity5 x 200μl
Titer>1 x 109 GC/ml
Vector MappAAV-G-CMV-2A-GFP
Gene NameHMGB1
SpeciesRat
AliasesAc2-008, Hmg1
Chromosome Location12p11
FormatVirus
Insert Size648
Vector Size5500
SerotypeAAV Serotype 1
Shipping ConditionsAAV viruses are shipped with dry ice. For long term storage, it is recommended to store the viruses at -80°C in small aliquots to avoid repeated freeze-thaw cycles.
SystemAAV Virus
Storage ConditionStore AAV viruses at -80°C. Avoid repeated freeze-thaw cycles by storing in small aliquots.
Guaranteeabm guarantees that the correct ORF construct is provided and the mRNA expression is displayed upon successful transduction. If this is not the case, we will provide a one-time replacement. Customers must provide adequate data to show >80% transfection efficiency with a positive control, plus additional qPCR data or a western blot to evaluate the level of gene expression. The replacement will not be covered by the same guarantee. Please note that due to the large number of variables applicable, any further expression analysis (e.g. protein expression) is not covered by the guarantee, as such analysis is dependent on the end user's experimental conditions.
Accession NumberNM_012963
FAQs


What is the difference between Retro-, Lenti-, and Adeno- viruses?
Retrovirus: Classic, can integrate into the genome but with low transduction efficiency. They are useful for gene transfer and protein expression in cells that have low transfection efficiency with other transfection reagents. Lentivirus: Can integrate into the genome with relatively high transduction efficiency and they are very useful for cells that have low transfection efficiency with other transfection reagents. No special competent cells required, as they are stable plasmids. Lentiviruses are a powerful tool for stable gene transfer to both dividing and non-dividing cells in vitro and in vivo. Adenovirus: Only work transiently (about 7 days) but have almost 100% transduction efficiency. Adenoviruses can infect a broad range of cell types with the highest efficiency and infection is not dependent on active host cell division. A second key feature is that high virus titers and high-level gene expression can be obtained in most mammalian cells.
What are the correct concentration units for each recombinant viral particle?
For lentiviruses and retroviruses, they are measured in CFU/ml (colony-forming units per millilitre). Transduction with lentiviruses and retroviruses can cause the formation of colonies, which can be quantified for concentration. For AAV the titer is measured as genome copies per mL (GC/mL). Adenoviruses are measured as PFU/ml (plaque-forming units per millilitre). Transduction with adenoviruses will kill packaging cells, forming plaques in the process for quantification. The concentration for all three types of viruses can also be classified as IU/ml (Infectious Units/ml). Ultimately, the units refers to the viral particles and different units reflect the different assays involved.
How long after transduction can the infection efficiency be observed?
You can observe transduction efficiency from 48 hours up to 5 days after infection.
How much DNA do I need to provide for Custom AAV without DNA Amplification?
You will need to provide purified plasmid DNA at a concentration of 0.5ug/ul or more.
50 ug DNA needed for Custom AAV without DNA amplification (10^9 GC/ml)
200 ug DNA needed for Custom AAV without DNA amplification (10^12 GC/ml)
300 ug DNA needed for Custom AAV without DNA amplification (10^13 GC/ml)
What's the difference between GC/ml and vg/ml?
Both are used interchangeably and it is a qPCR based titer method.
Why do I not see GFP expression after infecting AAV in my cell line?
Serotype selection is an important parameter affecting transduction ability of AAV particles. Thus, determine which serotype works best for your cell line. You could refer to our technical sheet "AAV - General Guideline to Serotype Selection". eg. Serotype 5 has limited transduction ability on most cell types.
Is your AAV preparation in-vivo grade?
Our high titer AAV preps undergo extensive purification steps leading to high quality viral particles ready to inject for your in-vivo models. For reference, see our in-vivo infectivity data as tested by an independent lab {https://www.abmgood.com/AAV-Adeno-Associated-Virus.html}
What are the QC methods for testing AAV?
We provide qPCR based titer as primary method of determining successful packaging. If your virus has GFP or RFP reporter, we also perform virus infectivity testing in HEK293T cell line and provide you image in CoA.Since infectivity is serotype dependent, and HEK293T cells are not infected well with Serotype 4,5 and 6, we are unable to confirm infection for these serotypes in our standard evaluation cell line. For such cases, titer as determined by qPCR is considered final parameter to determine successful packaging of virus.
Can I work with different serotypes of AAV virus in the same equipment to keep the infected cells?
There is no problem with using different serotypes in the same equipment, as long as the handler takes the basic precautions to avoid cross-contamination.
References


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  • Xu, T., He, B. S., Pan, B., Pan, Y. Q., Sun, H. L., Liu, X. X., ... & Wang, S. K. "MiR‐142‐3p functions as a tumor suppressor by targeting RAC1/PAK1 pathway in breast cancer" Journal of Cellular Physiology. : (2019).
  • Zhang, B., Roosmalen, I. A. M., Reis, C. R., Setroikromo, R., & Quax, W. J. "Death receptor 5 is activated by fucosylation in colon cancer cells" The FEBS Journal 286(3):555–571 (2019). DOI: 10.1111/febs.14742.
  • Zhao, Q., Zhao, S., Li, J., Zhang, H., Qian, C., Wang, H., … Zhao, Y. "TCF7L2 activated HOXA-AS2 decreased the glucocorticoid sensitivity in acute lymphoblastic leukemia through regulating HOXA3/EGFR/Ras/Raf/MEK/ERK pathway" Biomedicine & Pharmacotherapy 109:1640–1649 (2019). DOI: 10.1016/j.biopha.2018.10.046.