{"product_id":"araf-crispr-konckout-raw264-7-stable-cell-line-t1-19-bhc10900964","title":"ARAF CRISPR Konckout RAW264.7 Stable Cell Line-T1-19","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eThis product is a stable CRISPR knockout cell line in which \u003cem\u003eAraf\u003c\/em\u003e (A-RAF proto-oncogene, serine\/threonine kinase) has been disrupted in the RAW264.7 mouse macrophage background. The knockout enables loss-of-function investigation of ARAF's role in MAPK\/ERK signaling and innate immune responses within a macrophage model. RAW264.7 cells are a widely used murine macrophage-like cell line derived from an Abelson murine leukemia virus–induced tumor (Balb\/c, adult male). Cells grow as loosely adherent monolayers and are supplied frozen (BSL-II) with puromycin selection confirming stable knockout.\u003c\/p\u003e\n\n\u003ch2\u003eCRISPR Knockout Design\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eTarget gene:\u003c\/strong\u003e \u003cem\u003eAraf\u003c\/em\u003e (A-RAF proto-oncogene, serine\/threonine kinase; mouse gene symbol)\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eKnockout strategy:\u003c\/strong\u003e CRISPR-Cas9–mediated disruption of the \u003cem\u003eAraf\u003c\/em\u003e locus; stable integration confirmed by puromycin selection\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSelection marker:\u003c\/strong\u003e Puromycin resistance (add only after first passage post-thaw)\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eParental cell line:\u003c\/strong\u003e RAW264.7 — mouse macrophage cell line; adherent, loosely adherent; derived from ascites of an adult male Balb\/c mouse bearing an Abelson murine leukemia virus–induced tumor\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eClone ID:\u003c\/strong\u003e T1-19\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eGene Background\u003c\/h2\u003e\n\u003cp\u003eARAF (A-RAF proto-oncogene) is a member of the RAF family of serine\/threonine kinases, which also includes BRAF and CRAF (RAF1). RAF kinases are key effectors in the RAS–RAF–MEK–ERK mitogen-activated protein kinase (MAPK) cascade, linking extracellular growth factor signals to transcriptional regulation of cell proliferation, differentiation, and survival. While BRAF and CRAF have been more extensively characterized in cancer biology, ARAF is expressed in several tissues including macrophages and is involved in modulating MEK–ERK activation. In macrophages, MAPK signaling pathways including the ERK pathway are known to influence responses to inflammatory stimuli such as LPS. ARAF activity may contribute to the balance of inflammatory versus anti-inflammatory cytokine production.\u003c\/p\u003e\n\n\u003ch2\u003eCell Culture Specifications\u003c\/h2\u003e\n\u003cdl\u003e\n  \u003cdt\u003eGrowth properties\u003c\/dt\u003e\n\u003cdd\u003eAdherent, loosely adherent\u003c\/dd\u003e\n  \u003cdt\u003eOrganism\u003c\/dt\u003e\n\u003cdd\u003eMouse (\u003cem\u003eM. musculus\u003c\/em\u003e)\u003c\/dd\u003e\n  \u003cdt\u003eTissue origin\u003c\/dt\u003e\n\u003cdd\u003eAscites (macrophage; Abelson murine leukemia virus–induced tumor)\u003c\/dd\u003e\n  \u003cdt\u003eBiosafety level\u003c\/dt\u003e\n\u003cdd\u003eBSL-II\u003c\/dd\u003e\n  \u003cdt\u003eFormat\u003c\/dt\u003e\n\u003cdd\u003eFrozen; 1×10⁶ cells \/ 1.0 ml\u003c\/dd\u003e\n\u003c\/dl\u003e\n\u003cp\u003eKnockout is confirmed by the presence of frameshift-inducing INDELs, verified by Sanger Sequencing.\u003c\/p\u003e\n\n\u003ch2\u003eResearch Applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eMAPK\/ERK pathway analysis:\u003c\/strong\u003e Investigate the specific contribution of ARAF to MEK–ERK signaling in a macrophage context; compare phospho-ERK levels between Araf KO and parental RAW264.7 cells under basal and stimulated conditions.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eInnate immune signaling:\u003c\/strong\u003e Profile inflammatory cytokine production (e.g., TNFα, IL-6, IL-1β) in Araf-deficient vs. parental macrophages following LPS or other TLR stimulation.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eRAF isoform biology:\u003c\/strong\u003e Distinguish ARAF-dependent signaling from BRAF\/CRAF-mediated ERK activation using this KO line alongside isoform-specific inhibitors.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eDrug screening:\u003c\/strong\u003e Use the Araf KO line as a negative control when screening pan-RAF or ARAF-selective inhibitors to confirm on-target activity.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eMacrophage polarization:\u003c\/strong\u003e Assess M1 (pro-inflammatory) vs. M2 (anti-inflammatory) polarization capacity in ARAF-deficient RAW264.7 cells under appropriate stimulation conditions.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003ch2\u003eImportant Notes\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003eLoosely adherent growth:\u003c\/strong\u003e RAW264.7 cells are loosely adherent and may detach during routine media changes; use gentle aspiration and avoid disturbing the cell monolayer.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSelection drug timing:\u003c\/strong\u003e Add puromycin to culture medium only after the first passage post-thaw to allow cells to recover from freeze-thaw conditions.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eDonor\/source:\u003c\/strong\u003e RAW264.7 parental cells originate from ascites of an adult male Balb\/c mouse bearing an Abelson murine leukemia virus–induced tumor.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eViability warranty:\u003c\/strong\u003e Cell viability upon thaw is warranted for 30 days following shipment when handled according to abm's cell handling guidelines.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eIntended use:\u003c\/strong\u003e For laboratory research use only. Not intended for diagnostic, therapeutic, or clinical applications.\u003c\/li\u003e\n\u003c\/ul\u003e\n\n\u003c!-- Sources (internal):\n- ARAF gene entry | UniProt (P04798 mouse) | https:\/\/www.uniprot.org\/uniprot\/P04798\n- Araf gene record | NCBI Gene ID: 11836 | https:\/\/www.ncbi.nlm.nih.gov\/gene\/11836\n- Rapp UR et al. (1983) Structure and biological activity of v-raf, a unique oncogene transduced by a retrovirus. Proc Natl Acad Sci | https:\/\/doi.org\/10.1073\/pnas.80.14.4218\n- Dhomen N \u0026 Marais R (2007) New insight into BRAF mutations in cancer. Curr Opin Genet Dev | https:\/\/doi.org\/10.1016\/j.gde.2007.02.001\n- RAW264.7 ATCC entry | ATCC | https:\/\/www.atcc.org\/products\/tib-71\n--\u003e","brand":"Applied Biological Materials (abm) Inc.","offers":[{"title":"1x10\u003csup\u003e6\u003c\/sup\u003e cells \/ 1.0 ml","offer_id":53180708094317,"sku":"T9511","price":0.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/vROUbwTMp5UWYhp41hmtYYizSKrZlukIOYqpsiQo.png?v=1774960837","url":"https:\/\/www.ebiohippo.com\/products\/araf-crispr-konckout-raw264-7-stable-cell-line-t1-19-bhc10900964","provider":"BioHippo","version":"1.0","type":"link"}