NF-κB Luciferase Stable RAW264.7 Cell Line

Cat. No.
T3015
Unit
1x10⁶ cells / 1.0 ml
Price
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Cat. No. T3015
Name NF-κB Luciferase Stable RAW264.7 Cell Line
Description

NF-kB Luciferase Stable RAW264.7 Cell Line has been stably transfected with a 3kB-Luc SV40 reporter, which contains 3 NF-kB sites from the interferon gene upstream of the luciferase coding region. This cell line is useful in investigation of NF-kB activation in monocyte-macrophage models

Organism Mouse (M. musculus)
Tissue Ascites
Donor History Male, Balb/c, Adult, Abelson murine leukemia virus-induced tumor
Growth Properties Adherent, macrophage
Cell Type Stable Cell Lines
Unit 1x10⁶ cells / 1.0 ml
Storage Condition Vapor phase of liquid nitrogen, or below -130°C.
Shipping Conditions Ship with dry ice.
Product Format Frozen
Intended Use This product is intended for laboratory research use only. It is not intended for any animal or human therapeutic use, any human or animal consumption, or any diagnostic use.
BioSafety II
Certificate of Analysis For batch-specific test results, refer to the applicable certificate of analysis that can be found at www.abmgood.com.
Growth Conditions Use of PriCoat™ T25 Flasks (G299) or Applied Cell Extracellular Matrix (G422) is required for cell adhesion to the culture vessels. PriGrow I (TM001) + 10% FBS + 2 mM L-glutamine (G275) + 1% Penicillin/Streptomycin Solution (G255), 37.0°C, 5% CO₂. Selection with 100 µg/ml G418 (G271)
Unpacking and Storage Instructions

1. Visually examine the packaging containers for signs of leakage or breakage.

2. Immediately transfer frozen cells from dry ice packaging to a temperature below -130°C, preferably in liquid nitrogen vapor phase storage, until ready for use.

To ensure the highest level of viability, thaw the vial and initiate culture as soon as possible upon receipt. If continued storage is desired, the vial should only be stored below -130°C or in liquid nitrogen vapor phase. Do not store at -70°C, as it will result in loss of viability.


Thawing Protocol

1. Thaw cells quickly in a 37°C water bath while agitating gently (maximum 2 minutes). The vial cap should be kept above the water level to minimize the risk of contamination.

2. Decontaminate the vial by spraying and wiping the exterior of the vial with 70% ethanol. From this point onwards, all operations should be strictly carried out inside a biological safety cabinet using aseptic conditions.

3. Transfer the cell suspension into a 15ml sterile conical tube containing 5ml of pre-warmed, complete growth media. Centrifuge cells at 125xg for 5-7 minutes.

4. Aspirate the supernatant without disturbing the cell pellet. Re-suspend the cell pellet in the recommended pre-warmed, complete growth media and dispense into a T25 culture flask.

5. Incubate the cells at the recommended conditions.

Subculture Protocol

Volumes given below are for a T75 flask; proportionally increase or decrease the volume as required per culture vessel size. Subculture cells once the culture vessel is 80% confluent.

1. Aspirate the culture media, and add 2-3ml of pre-warmed 0.25% Trypsin-EDTA to the culture vessel.

2. Observe the cells under a microscope to confirm detachment (typically within 2-10 minutes). Cells that are difficult to detach can be put in 37°C, for several minutes to facilitate detachment.

3. Neutralize Trypsin-EDTA by adding an equal volume of the complete growth media into the culture vessel.

4. Transfer the culture suspension into a sterile centrifuge tube, and centrifuge at 125xg for 5 minutes. The actual centrifuge duration and speed may vary depending on the cell type.

5. Aspirate the supernatant, and re-suspend the pellet with pre-warmed fresh complete growth media. Add appropriate aliquots of the cell suspension to new culture vessels, as desired.

6. Incubate the cells at the recommended conditions.

Cryopreservation Cryopreservation Medium (TM024), or complete growth media with 10% DMSO.
Seeding Density (cells/cm2) 50,000 - 70,000
Split Ratio 1:5
Population Doubling Time (h) 30 - 35
Expression NGF, Iba1, TREM2, CD11b, CD68, SV40 (detected via RT-PCR)
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. T3015.
Warranty 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”.
Disclaimer

1. Sale of this item is subjected to the completion of a Material Transfer Agreement (MTA) by the purchasing individual/institution for each cell line. If you have any questions regarding this, please contact us at [email protected].

2. All test parameters provided in the CoA are conducted using abm's standardized culture system and 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 μg, Cat.# C207, $450.00) or cell lysate (100 μg, Cat.# C206, $600.00) to perform preliminary experiments, or alternatively use our Gene Expression Assay Service (Cat# C138). All sales are final.

3. We recommend live cell shipments for ease of cell transfer and this option can be requested at the time of order placement. 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.

4. 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 or any other non-RUO application(s).

5. abm makes no warranties or representations as to the accuracy of the information on this site. Citations from literature are provided for informational purposes only. abm does not warrant that such information has been shown to be accurate.

6. 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".

Depositor University of Western Australia
Application Research Use 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. T3015
Print & Download Datasheet
  • Wang, C et al. "12-O-tetradecanoylphorbol-13-acetate (TPA) inhibits osteoclastogenesis by suppressing RANKL-induced NF-kB activation" J Bone Miner Res 18:2159-2168 (2003). DOI: 10.1359/jbmr.2003.18.12.2159.


    Xu, J et al. "Evidence of reciprocal regulation between the high extracellular calcium and RANKL signal transduction pathways in RAW cell derived osteoclasts" J Cell Physiol. 202(2):554-62 (2005). PubMed: 15389575.

    King, TJ et al. "Methotrexate chemotherapy promotes osteoclast formation in the long bone of rats via increased pro-inflammatory cytokines and enhanced NF-κB activation" Am J Pathol. 181(1):121-9 (2012). DOI: 10.1016/j.ajpath.2012.03.037. PubMed: 22642908.


    Singh, PP et al. "Membrane-bound receptor activator of NFκB ligand (RANKL) activity displayed by osteoblasts is differentially regulated by osteolytic factors" Biochem Biophys Res Commun 422(1):48-53 (2012). DOI: 10.1016/j.bbrc.2012.04.103.


    Li, X et al. "Sinomenine suppresses osteoclast formation and Mycobacterium tuberculosis H37Ra-induced bone loss by modulating RANKL signaling pathways" PLoS One 8(9):e74274 (2013). DOI: 10.1371/journal.pone.0074274.


    Liu, Q et al. "SC-514, a selective inhibitor of IKKβ attenuates RANKL-induced osteoclastogenesis and NF-κB activation" Biochem Pharmacol. 86(12):1775-83 (2013). DOI: 10.1016/j.bcp.2013.09.017. PubMed: 24091016.


    van der Kraan, AG et al. "HSP90 inhibitors enhance differentiation and MITF (microphthalmia transcription factor) activity in osteoclast progenitors" Biochem J. 451(2):235-44 (2013). DOI: 10.1042/BJ20121626. PubMed: 23379601.


    Hirai, K., Furusho, H., Hirota, K., & Sasaki, H. (2018). Activation of hypoxia-inducible factor 1 attenuates periapical inflammation and bone loss. International journal of oral science, 10(2), 12. https://doi.org/10.1038/s41368-018-0015-0


    de Camargo, A. C., Biasoto, A. C. T., Schwember, A. R., Granato, D., Rasera, G. B., Franchin, M., ... & Shahidi, F. (2019). Should we ban total phenolics and antioxidant screening methods? The link between antioxidant potential and activation of NF-κB using phenolic compounds from grape by-products. Food chemistry, 290, 229-238.


    Lazarini, J. G. (2020). Phytochemical, toxicity, and evaluation of anti-inflammatory and antioxidant activities of Eugenia neonitida Sobral (pitangatuba), a Brazilian native fruit: Fitoquímica, toxicidade e avaliação das atividades anti-inflamatória e Sobral (pitangatuba), uma fruta nativa do Brasil.

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