E1A-E1B Lentivirus, High Titer
| Cat. No. | LV652 |
| Name | E1A-E1B Lentivirus, High Titer |
| Unit | 2 x 100 µl |
| Unpacking and Storage Instructions |
Lentiviruses 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. |
| Description |
High Titer (109 IU/ml) Recombinant Lentivirus expressing the E1A-E1B gene. |
| Application |
Immortalization of primary rodent cells, including amniotic fluid cells, primary epithelial cells, fibroblasts, and primary kidney cells. |
| Expression System Type | Lentivirus |
| Caution |
This product is distributed for laboratory research only. |
| Material Citation | If use of this material results in a scientific publication, please cite the material in the following manner: Applied Biological Materials Inc, Cat. No. LV652 |
Print/Download Datasheet
| How are endotoxin levels measured? | |
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1. For estimating the endotoxin levels; we use the LAL (Limulus Amebocyte Lysate) method: The lysate from horseshoe crab amebocytes clots in the presence of very low endotoxin. This reaction is the basis of the Limulus amebocyte lysate (LAL) assay which was approved by the FDA in 1970.
· Endotoxin is generally measured in Endotoxin Units per milliliter (EU/mL).
· For recombinant proteins: EU is reported per microgram of protein.
· One EU = 0.1-0.2 ng endotoxin/µg of protein.
· At abm, we do the LAL chromogenic assays that can detect down to 0.01 EU/ml.
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| With regard to the BSA levels in some Growth Factors and Cytokines, can you please provide an explanation as to why they are so high? | |
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The amount of BSA, as part of the formulation of a protein, can vary considerably depending on how much BSA was deemed optimum/necessary for protein stability in combination with /in-lieu of - other possible additives. The aforementioned formulations are somewhat analogous to the “carrier” versions of many formulations from “R and D systems” that have as high as 50 µg of BSA per µg of the recombinant protein product. If, needed or desired, abm scientists can substitute BSA for other stabilizing additives for most formulations.
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| Are your Escherichia coli sourced growth factors: 1) Human derived materials free? 2) Recombinant proteins free? | |
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Yes, all of abm's growth factors made in Escherichia coli using recombinant technology contain no human derived-products or other recombinant proteins. In the rare cases of BSA presence, this will be mentioned in the product's formulation.
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| What is the general procedure for immortalizing cells? | |
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See our Cell Immortalization Handbook for more details.
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| Can you recommend a specific reagent and protocol to immortalize my cell line? | |
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Successful cell immortalization is unpredictable and must be determined experimentally, as results vary with species, tissue origin, growth characteristics and whether or not specific cell functions need to be preserved.
Our in-house experiments using the SV40T and hTERT lentiviruses have achieved the highest immortalization success rates in a variety of different cell types. |
| Do immortalized cells retain all characteristics of the original primary cell line? | |
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Not always. Some phenotypic or genetic changes may occur during immortalization. We recommend using our hTERT immortalization reagents as this method preserves phenotype and karyotype as well as minimizes genomic instability.
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| Do I need a control when performing cell immortalization experiments? | |
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Yes. Always include control cells that are not transduced in order to compare growth rates and detect senescence bypass.
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| How do I confirm successful cell immortalization? | |
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Assess long-term growth beyond normal passage limits and verify expression of the immortalizing genes via PCR, qPCR or Western blot.
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| Where can I learn more about cell immortalization? | |
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Check out our Learning Resources and Youtube video for more information.
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| Will this virus immortalize my primary cells? | |
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This is highly dependent on the immortalization reagent and cell type. Check out our Cell Immortalization Reagents Compatibility Chart for more information.
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