Phospho-RAF1 (S259) Recombinant Monoclonal Antibody

說明書

中文名稱：

磷酸化-RAF1 (S259)重組抗體
貨號：

CSB-RA019284A259phHU
規格：

￥1320
圖片：
Western Blot
Positive WB detected in Hela whole cell lysate(treated with Calyculin A or EGF)
All lanes Phospho-RAF1 antibody at 1.18μg/ml
Secondary
Goat polyclonal to rabbit IgG at 1/50000 dilution
Predicted band size: 73 KDa
Observed band size: 73 KDa

IHC image of CSB-RA019284A259phHU diluted at 1:100 and staining in paraffin-embedded human endometrial cancer performed on a Leica BondTM system. After dewaxing and hydration, antigen retrieval was mediated by high pressure in a citrate buffer (pH 6.0). Section was blocked with 10% normal goat serum 30min at RT. Then primary antibody (1% BSA) was incubated at 4℃ overnight. The primary is detected by a biotinylated secondary antibody and visualized using an HRP conjugated SP system.

Immunofluorescence staining of HepG2 cells with CSB-RA019284A259phHU at 1:100,counter-stained with DAPI. The cells were fixed in 4% formaldehyde, permeabilized using 0.2% Triton X-100 and blocked in 10% normal Goat Serum. The cells were then incubated with the antibody overnight at 4℃. The secondary antibody was Alexa Fluor 488-congugated AffiniPure Goat Anti-Rabbit IgG (H+L).
其他：

產品詳情
產品評價
相關產品
靶點詳情

產品詳情

產品描述：

CSB-RA019284A259phHU是一種靶向RAF1蛋白S259磷酸化位點的重組單克隆抗體，適用于檢測RAF1在該關鍵位點的翻譯后修飾狀態。RAF1作為MAPK/ERK信號通路的核心成員，通過S259位點的磷酸化參與調控自身激酶活性及亞細胞定位，對細胞增殖、分化及應激響應等生物學過程具有重要影響。該抗體經嚴格驗證，在多種實驗場景中展現出優異的特異性和靈敏度：在Western Blot中可清晰識別內源性磷酸化RAF1蛋白，推薦稀釋范圍為1:500-1:5000；免疫組化（IHC）和免疫熒光（IF）實驗分別適用于組織切片及細胞樣本的亞細胞定位分析，推薦使用稀釋度為1:50-1:200和1:20-1:200。其重組單克隆特性確保了批次間的高度一致性，適用于研究RAF1在腫瘤發生、耐藥機制以及發育調控中的動態磷酸化過程，為探索RAS/MAPK通路異常相關疾病的分子機制提供可靠工具。該產品嚴格限于科學研究領域，可配合多種模型系統進行蛋白功能研究及信號轉導網絡解析。
Uniprot No.：

P04049
基因名：

RAF1
別名：

c Raf antibody; C-raf antibody; C-Raf proto-oncogene, serine/threonine kinase antibody; CMD1NN antibody; Craf 1 transforming gene antibody; cRaf antibody; Craf1 transforming gene antibody; EC 2.7.11.1 antibody; kinase Raf1 antibody; Murine sarcoma 3611 oncogene 1 antibody; NS5 antibody; Oncogene MIL antibody; Oncogene RAF1 antibody; OTTHUMP00000160218 antibody; OTTHUMP00000207813 antibody; OTTHUMP00000209389 antibody; Protein kinase raf 1 antibody; Proto-oncogene c-RAF antibody; Raf 1 antibody; Raf 1 proto oncogene serine/threonine kinase antibody; RAF antibody; Raf proto oncogene serine/threonine protein kinase antibody; RAF proto-oncogene serine/threonine-protein kinase antibody; RAF-1 antibody; RAF1 antibody; RAF1_HUMAN antibody; Similar to murine leukemia viral (V-raf-1) oncogene homolog 1 antibody; TRANSFORMING REPLICATION-DEFECTIVE MURINE RETROVIRUS 3611-MSV antibody; v raf 1 murine leukemia viral oncogene homolog 1 antibody; v-raf murine sarcoma viral oncogene homolog 1 antibody; v-raf-1 murine leukemia viral oncogene-like protein 1 antibody; vraf1 murine leukemia viral oncogene homolog 1 antibody
反應種屬：

Human
免疫原：

A synthesized peptide derived from Human Phospho-RAF1 (S259)
免疫原種屬：

Homo sapiens (Human)
標記方式：

Non-conjugated
克隆類型：

Monoclonal
抗體亞型：

Rabbit IgG
純化方式：

Affinity-chromatography
克隆號：

4C9
濃度：

It differs from different batches. Please contact us to confirm it.
保存緩沖液：

Rabbit IgG in 10mM phosphate buffered saline , pH 7.4, 150mM sodium chloride, 0.05% BSA, 0.02% sodium azide and 50% glycerol.
產品提供形式：

Liquid
應用范圍：

ELISA, WB, IHC, IF

推薦稀釋比：

Application	Recommended Dilution
WB	1:500-1:5000
IHC	1:50-1:200
IF	1:20-1:200

Protocols：

ELISA Protocol
Western Blotting (WB) Protocol
Immunohistochemistry (IHC) Protocol
Immunofluorescence (IF) Protocol
儲存條件：

Upon receipt, store at -20°C or -80°C. Avoid repeated freeze.
貨期：

Basically, we can dispatch the products out in 1-3 working days after receiving your orders. Delivery time maybe differs from different purchasing way or location, please kindly consult your local distributors for specific delivery time.
用途：

For Research Use Only. Not for use in diagnostic or therapeutic procedures.

產品評價

靶點詳情

功能：

Serine/threonine-protein kinase that acts as a regulatory link between the membrane-associated Ras GTPases and the MAPK/ERK cascade, and this critical regulatory link functions as a switch determining cell fate decisions including proliferation, differentiation, apoptosis, survival and oncogenic transformation. RAF1 activation initiates a mitogen-activated protein kinase (MAPK) cascade that comprises a sequential phosphorylation of the dual-specific MAPK kinases (MAP2K1/MEK1 and MAP2K2/MEK2) and the extracellular signal-regulated kinases (MAPK3/ERK1 and MAPK1/ERK2). The phosphorylated form of RAF1 (on residues Ser-338 and Ser-339, by PAK1) phosphorylates BAD/Bcl2-antagonist of cell death at 'Ser-75'. Phosphorylates adenylyl cyclases: ADCY2, ADCY5 and ADCY6, resulting in their activation. Phosphorylates PPP1R12A resulting in inhibition of the phosphatase activity. Phosphorylates TNNT2/cardiac muscle troponin T. Can promote NF-kB activation and inhibit signal transducers involved in motility (ROCK2), apoptosis (MAP3K5/ASK1 and STK3/MST2), proliferation and angiogenesis (RB1). Can protect cells from apoptosis also by translocating to the mitochondria where it binds BCL2 and displaces BAD/Bcl2-antagonist of cell death. Regulates Rho signaling and migration, and is required for normal wound healing. Plays a role in the oncogenic transformation of epithelial cells via repression of the TJ protein, occludin (OCLN) by inducing the up-regulation of a transcriptional repressor SNAI2/SLUG, which induces down-regulation of OCLN. Restricts caspase activation in response to selected stimuli, notably Fas stimulation, pathogen-mediated macrophage apoptosis, and erythroid differentiation.
基因功能參考文獻：
1. The functional assessment supported the pathogenicity of the RAF1 and RIT1 VUSs, while the significance of two variants of unknown significance in A2ML1 remained unclear. PMID: 29402968
2. Our report presents the second familial case of Noonan syndrome due to a germline p.S427G substitution in RAF] with no occurrence of a malignant tumor. It may suggest that carrying a germline mutation in the RAF1 oncogene is not associated with an increased risk of tumor development. Since RAF1 mutations have been observed as a somatic event in many types of cancer. PMID: 30204961
3. Data indicate that Raf-1 proto-oncogene, serine-threonine kinase (RAF1) is a negative regulator of hepatocarcinogenesis. PMID: 28000790
4. we report a patient with an inherited RAF1-associated Noonan syndrome, presenting with an antenatally diagnosed abnormality of skull shape, bilateral subdural haematomas, of unknown cause, delayed myelination and polymicrogyria. PMID: 27753652
5. Raf1 may serve as a novel prognostic factor and potential target for improving the longterm outcome of nonsmall cell lung cancer (NSCLC). PMID: 29484414
6. Results provide evidence that RAF1 binding to SPRY4 is regulated by miR-1908 in glioma tumors. PMID: 29048686
7. High RAF1 expression is associated with malignant melanoma. PMID: 28677804
8. two premature neonates with progressive biventricular hypertrophy found to have RAF1 variants in the CR2 domain, are reported. PMID: 28777121
9. Data indicate connector enhancer of kinase suppressor of Ras 1 protein (CNK1) as a molecular platform that controls c-raf protein (RAF) and c-akt protein (AKT) signalling and determines cell fate decisions in a cell type- and cell stage-dependent manner. PMID: 27901111
10. CRAF is a bona fide alternative oncogene for BRAF/NRAS/GNAQ/GNA11 wild type melanomas PMID: 27273450
11. Authors evaluated the expression of known targets of miR-125a and found that sirtuin-7, matrix metalloproteinase-11, and c-Raf were up-regulated in tumor tissue by 2.2-, 3-, and 1.7-fold, respectively. Overall, these data support a tumor suppressor role for miR-125a. PMID: 28445974
12. Overexpression of ciRS-7 in HCT116 and HT29 cells led to the blocking of miR-7 and resulted in a more aggressive oncogenic phenotype, and ciRS-7 overexpression permitted the inhibition of miR-7 and subsequent activation of EGFR and RAF1 oncogenes PMID: 28174233
13. miR-497 could serve as a tumor suppressor and a potential early diagnostic marker of gastric cancer by targeting Raf-1 proto-oncogene. PMID: 28586056
14. RAF1 may have a role in survival in hepatocellular carcinoma, and indicate whether sorafenib should be used as a postoperative adjuvant PMID: 26981887
15. Mutational activation of Kit-, Ras/Raf/Erk- and Akt- pathways indicate the biological importance of these pathways and their components as potential targets for therapy. PMID: 27391150
16. Results indicate that des-gamma-carboxy prothrombin (DCP) antagonizes the inhibitory effects of Sorafenib on hepatocellular carcinoma (HCC) through activation of the Raf/MEK/ERK and PI3K/Akt/mTOR signaling pathways. PMID: 27167344
17. DiRas3 binds to KSR1 independently of its interaction with activated Ras and RAF. PMID: 27368419
18. RhoA/ROCK and Raf-1/CK2 pathway are responsible for TNF-alpha-mediated endothelial cytotoxicity via regulation of the vimentin cytoskeleton. PMID: 28743511
19. Although Raf-1 gene is not mutated, an abnormality of Raf-1 kinase feedback regulation enhances its antiapoptotic function, and Raf-1 can still be a pharmaceutical target to increase chemotherapy or radiotherapy sensitivity in these cancer cells. PMID: 27841865
20. RAF1 plays a critical role in maintaining the transformed phenotype of CRC cells, including those with mutated KRAS. PMID: 27670374
21. This finding suggests that stringent assemblage of Hsp90 keeps CRAF kinase equipped for participating in the MAPK pathway. Thus, the role of Hsp90 in CRAF maturation and activation acts as a limiting factor to maintain the function of a strong client like CRAF kinase. PMID: 27703006
22. Oncogenic NFIA:RAF1 fusion activation of the MAPK pathway is associated with pilocytic astrocytoma. PMID: 27810072
23. IGF2BP2 as a post-transcriptional regulatory mRNA-binding factor, interfering with Raf-1 degradation by miR-195, that contributes to Colorectal carcinogenesis. PMID: 27153315
24. Data show that when microRNA miR-125b was over-expressed in THP-1 macrophages, the expression of Raf1 proto-oncogene serine/threonine protein kinase (RAF1) was reduced to promote the apoptosis of macrophages. PMID: 27363278
25. Data show that Griffipavixanthone (GPX), a dimeric xanthone isolated from Garcinia esculenta, is a B-RAF and C-RAF inhibitor against esophageal cancer cells. PMID: 26646323
26. Up-regulation of Raf-1 is associated with triple-negative breast cancer. PMID: 26513016
27. This study provides the molecular basis for C-Raf C-terminal-derived phosphopeptide interaction with 14-3-3zeta protein and gives structural insights responsible for phosphorylation-mediated protein binding. PMID: 26295714
28. a model that CD166 regulates MCAM through a signaling flow from activation of PI3K/AKT and c-Raf/MEK/ERK signaling to the inhibition of potential MCAM ubiquitin E3 ligases, betaTrCP and Smurf1. PMID: 26004137
29. Suggest an interrelated kinase module involving c-Raf/PI3K/Lyn and perhaps Fgr functions in a nontraditional way during retinoic acid-induced maturation or during rescue of RA induction therapy using inhibitor co-treatment in RA-resistant leukemia cells. PMID: 25817574
30. Abnormal activation of the Ras/MAPK pathway may play a significant role in the development of pulmonary vascular disease in the subset of patients with Noonan syndrome and a specific RAF1 mutation. PMID: 25706034
31. Raf-1 may be an important biomarker in predicting the prognosis of chordoma patients. PMID: 25755752
32. In the presence of Raf1, the RasQ61L mutant has a rigid switch II relative to the wild-type and increased flexibility at the interface with switch I, which propagates across Raf-Ras binding domain. PMID: 25684575
33. Besides mediating the anticancer effect, pDAPK(S308) may serve as a predictive biomarker for Raf inhibitors combination therapy, suggesting an ideal preclinical model that is worthy of clinical translation. PMID: 26100670
34. DJ-1 directly binds to the kinase domain of c-Raf to stimulate its self-phosphorylation, followed by phosphorylation of MEK and ERK1/2 in EGF-treated cells. PMID: 26048984
35. truncated RAF1 and BRAF proteins, recently described as products of genomic rearrangements in gastric cancer and other malignancies, have the ability to render neoplastic cells resistant to RTK-targeted therapy PMID: 25473895
36. our study demonstrated that miR-455-RAF1 may represent a new potential therapeutic target for colorectal carcinoma treatment. PMID: 25355599
37. approach identified 18 kinase and kinase-related genes whose overexpression can substitute for EGFR in EGFR-dependent PC9 cells, and these genes include seven of nine Src family kinase genes, FGFR1, FGFR2, ITK, NTRK1, NTRK2, MOS, MST1R, and RAF1. PMID: 25512530
38. Aberrant expression of A-, B-, and C-RAF, and COT is frequent in PTC; increased expression of COT is correlated with recurrence of PTC. PMID: 25674762
39. Authors demonstrate that the N-terminus of human Raf1 kinase (hRaf11-147aa) binds with human RKIP (hRKIP) at its ligand-binding pocket, loop "127-149", and the C-terminal helix by nuclear magnetic resonance experiments. PMID: 24863296
40. Including several anti-apoptotic Bcl-2 family members and c-Raf. PMID: 24969872
41. These data suggest that miR-7-5p functions as a tumor suppressor gene to regulate glioblastoma microvascular endothelial cell proliferation potentially by targeting the RAF1 oncogene PMID: 25027403
42. A novel mechanism for response was discovered whereby high expression level of CAV-1 at the plasma membrane disrupts the BRaf/CRaf heterodimer and thus inhibits the activation of MAPK pathway during dasatinib treatment. PMID: 24486585
43. Results show that ubiquitination and levels of RAF-1 is controlled by both Shoc2 and HUWE1. PMID: 25022756
44. Raf-1/JNK /p53/p21 pathway may be involved in apoptosis, and NFkappaB1 may play a possible role in inhibiting apoptosis. PMID: 22282237
45. The higher expression of RAF1 mRNA and the activation of AKT/ERK proteins in vinorelbine-resistant non-small cell lung cancer cell lines may confer resistance to vinorelbine PMID: 24427333
46. analysis of RAF1 mutations in cohorts of South Indian, North Indian and Japanese patients with childhood-onset dilated cardiomyopathy PMID: 24777450
47. Expression of miR-195 or knockdown of Raf-1 can similarly reduce tumor cell survival. PMID: 23760062
48. We hypothesize a potential direct or indirect role for SRC, RAF1, PTK2B genes in neurotransmission and in central nervous system signaling processes. PMID: 24108181
49. we identified multiple C-RAF mutations that produced biochemical and pharmacologic resistance in melanoma cell lines PMID: 23737487
50. ARAF seems to stabilize BRAF:CRAF complexes in cells treated with RAF inhibitors and thereby regulate cell signaling in a subtle manner to ensure signaling efficiency PMID: 22926515
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相關疾病：

Noonan syndrome 5 (NS5); LEOPARD syndrome 2 (LPRD2); Cardiomyopathy, dilated 1NN (CMD1NN)
亞細胞定位：

Cytoplasm. Cell membrane. Mitochondrion. Nucleus. Note=Colocalizes with RGS14 and BRAF in both the cytoplasm and membranes. Phosphorylation at Ser-259 impairs its membrane accumulation. Recruited to the cell membrane by the active Ras protein. Phosphorylation at Ser-338 and Ser-339 by PAK1 is required for its mitochondrial localization. Retinoic acid-induced Ser-621 phosphorylated form of RAF1 is predominantly localized at the nucleus.
蛋白家族：

Protein kinase superfamily, TKL Ser/Thr protein kinase family, RAF subfamily
組織特異性：

In skeletal muscle, isoform 1 is more abundant than isoform 2.
數據庫鏈接：

HGNC: 9829

OMIM: 164760

KEGG: hsa:5894

STRING: 9606.ENSP00000251849

UniGene: Hs.159130

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