H295RA is an ACTH-responsive human adrenocortical cell line, produced from H295R cells that have been transduced to stable express increased human MRAP (MC2R accessory protein) gene. H295R adrenocortical cell line displays relatively low ACTH responsiveness due to low melanocortin 2 receptor (MC2R) expression. Successful induction of MRAP and increased expression of MC2R in H295RA cells was confirmed by quantitative real-time RT-PCR and protein analysis. This cell line is well-suited for research regarding molecular analysis of physiologic and pathologic conditions involving the hypothalamicpituitaryadrenal axis.
Cell Type
Tumor Cells
Disclaimer
1. For for-profit organizations and corporations, please contact orders@biohippo.com for pricing of this item. 2. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each order. If you have any questions regarding this, please contact us at licensing@abmgood.com. 3. All test parameters provided in the CoA are conducted using abm's standardized culture system and procedures. The stated values may vary under the end-user's culture conditions. Please verify that the product is suitable for your studies by referencing published papers or ordering RNA (0.5 mug, Cat.# C207, $450.00) or cell lysate (100 mug, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final. 4. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of ordering. Please note that the end-user will need to evaluate the feasibility of live cell shipment by taking into account the final destination's temperature variation and its geographical location. In addition, we thoroughly test our cell lines for freeze-thaw recovery. If frozen cells were received and not recovered in your lab under the exact, specified conditions (using recommended culture vessel, media, additional supplements, and atmospheric conditions), a live cell replacement is possible at a cost (plus shipping). 5. All of abm's cell biology products are for research use ONLY and NOT for therapeutic/diagnostic applications. abm is not liable for any repercussions arising from the use of its cell biology product(s) in therapeutic/diagnostic application(s). Please contact a technical service representative for more information. 6. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature and provided for informational purposes only. abm does not warrant that such information has been shown to be accurate. 7. abm warrants that cell lines shall be viable upon initiation of culture for a period of thirty (30) days after shipment and that they shall meet the specifications on the applicable abm Material Product Information sheet, certificate of analysis, and/or catalog description. Such thirty (30) day period is referred to herein as the "Warranty Period."
Expression Level
Transgenic expression of human MRAP; ACTH responsive steroid production and enzyme induction; angiotensin II responsive aldosterone production and aldosterone synthase expression. Selection marker: blasticidin 1ug/mL. Vector: pLX 304-Blast-V5 (Accession #BC062721)
Quantity
1x106 cells / 1.0 ml
Tissue
Renal
Morphology
Polygonal
Population Doubling
36 - 48 hours
Species
Human (H. sapiens)
Propagation
Use of PriCoatTM T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. Grow cells in ECM-coated culture vessels unless otherwise specified in the Propagation Requirements below.The base medium for this cell line is Prigrow IV medium available at abm, Cat. No. TM004. To make the complete growth medium, add the following components to the base medium: cosmic calf serum (Hyclone) to a final concentration of 10%, Penicillin/Streptomycin Solution (G255) to a final concentration of 1%, ITS Plus (BD Biosciences) to a final concentration of 1%, and Gentamicin (G272) to a final concentration of 1 mug/mL. Cells may be grown in the presence of 1 mug/mL blasticidin (Thermo Fisher).Change media every 2-3 days.Carbon dioxide (CO2): 5%, Temperature: 37.0°C.
Quality Control
1) Monoclonal selection using cloning cylinders; 2) quantitative real-time PCR; 3) Protein analysis