Product name :E1B

Catalog Number :000008A

Quantity :250ul

Price :1065.75 Eur

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Supplier :ABM Goods

Vector pAdeno
Promoter CMV
Insert E1B
Organism Human
Accession # P21953
Alias BCKDHB
Protein Type Metabolic
Protein Family Unknown
Functions BCKDH-Beta catalyzes the second major step in the catabolism of the branched-chain amino acids, isoleucine, leucine and valine.
Cell Types Unknown
Protein Location Mitochondrial matrix
Chromosome 6p22-p21
Protein Structure BCKDH-Beta contains two isoforms, having 392 amino acids each respectively.
Gene Type Mutant
Titer 1x106pfu/mL
Storage DMEM with 2.5% glycerol
Caution This product is for research use only and is not intended for therapeutic or diagnostic applications. Please contact a technical service representative for more information.

Protocol:

Always amplify one adenovirus at a time and in different culture hoods and incubators. If
only one set of equipment is available, amplify the viruses sequentially and use UV radiation
for 30 minutes in‐between each virus. Use separate trypsin and medium containers for each
virus. Cross‐contamination when working with two or more adenoviruses is more common
than you think. Once it occurred, your results will be greatly compromised. After you have
individual stocks for each adenovirus, you can then work with two or more adenoviruses in
your targeting cells.
Furthermore, large‐scale virus production and purification is necessary for in vivo injection
and most in vitro gene transductions.

1. When you receive your recombinant adenovirus, make two to three aliquots and use one
for amplification in 293 cells. Freeze the other aliquots in ‐70°C as a seed stock for
future use.
2. Amplify your adenovirus in 293 cells by infecting the cells with 10μL of the adenovirus
for a 60mm dish, or 200μL for a 100mm dish.
3. When more than 95% of 293 cells are detached from dishes, collect both cells and
medium.
4. Freeze (‐70°C freezer or dry ice / ethanol) and thaw (37°C water bath) the collection three
times.
5. Pellet cell debris by centrifugation at 3,000 rpm at room temperature for 10 minutes.
6. Transfer supernatant into a new tube. Store at 4°C for short‐term use (two to three weeks)
or add glycerol (final concentration 10%) and freeze at ‐70°C (stable for one to two years).
7. Use the supernatant to infect your target cells. Subsequently analyze your gene
expression by Western blotting, Q‐PCR, or under microscope if your gene of interest is a
reporter gene (i.e. β‐gal or EGFP).
8. For any further questions, please contact us at info@abmGood.com and we will get back
to you promptly.

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