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RAF1 抗体 (AA 31-130)

RAF1 适用: 人 ELISA, IHC (p), IF (cc), IF (p), IHC (fro) 宿主: 兔 Polyclonal unconjugated
产品编号 ABIN733208
发货至: 中国
  • 抗原 See all RAF1 抗体
    RAF1 (V-Raf-1 Murine Leukemia Viral Oncogene Homolog 1 (RAF1))
    抗原表位
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    AA 31-130
    适用
    • 447
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    宿主
    • 478
    • 15
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    克隆类型
    • 453
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    多克隆
    标记
    • 202
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    This RAF1 antibody is un-conjugated
    应用范围
    • 298
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    ELISA, Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunofluorescence (Cultured Cells) (IF (cc)), Immunofluorescence (Paraffin-embedded Sections) (IF (p)), Immunohistochemistry (Frozen Sections) (IHC (fro))
    交叉反应
    预测反应
    Mouse,Rat,Dog,Cow,Horse,Rabbit
    纯化方法
    Purified by Protein A.
    免疫原
    KLH conjugated synthetic peptide derived from human c Raf
    亚型
    IgG
  • 应用备注
    ELISA 1:500-1000
    IHC-P 1:200-400
    IHC-F 1:100-500
    IF(IHC-P) 1:50-200
    IF(IHC-F) 1:50-200
    IF(ICC) 1:50-200
    限制
    仅限研究用
  • 生效 #029651 (Immunohistochemistry)
    '独立验证'标志
    by
    Immunohistochemistry Core, NYU Langone
    No.
    #029651
    日期
    2014.04.01
    抗原
    Lot Number
    130909
    Method validated
    Immunohistochemistry
    Negative Control
    Human liver
    Notes
    Raf1 exhibits uniformly low expression across all human tissues; higher expression found in colon, lower in liver. Tissue-specific expression is detected in colon, with expected low level expression in liver.
    '独立验证'标志
    Validation Images
    Full Methods
    Primary Antibody
    • Antigen: V-Raf-1 Murine Leukemia Viral Oncogene Homolog 1 (RAF1)
    • Catalog number: ABIN733208
    • Lot number: 130909
    Secondary Antibody
    • Lot number: D07640BA
    Full Protocol
    • Immunohistochemistry was performed on a Ventana NEXes automated platform; instrument manufacturer specific reagents are italicized.
    • 1. Slides were preheated in convection oven at 60°C for 30 min
    • 2. Deparaffinization procedure: - 3 changes of Xylene, 5 min each - 3 changes of 100% Ethanol, 3 min each - 3 changes of 95% Ethanol, 3 min each - Rinsed in distilled water, 3 changes
    • 3. Heat retrieval procedure - Slides retrieved in 10.0 mM Citrate, pH6.0 in a 1000W microwave oven (~100°C) for 15 min. - Slides were allowed to cool (in citrate) for 30 min. - Slides were washed x 3 in Distilled water
    • 4. NEXes instrument procedure, iView DAB paraffin protocol (*abridged*): - Slide chamber warmed to 37°C
    • 5. Slides rinsed with *reaction buffer* x3
    • 6. *iView Inhibitor (H2O2)* applied and incubated for 4 min
    • 7. Slides rinsed with *reaction buffer*
    • 8. Antibody Application - Primary antibody diluted 1:250 in PBS (100 microliter applied/slide) - Ventana Isotype control applied neat - Slides Incubated overnight at room temperature (~12 hours ~25°C)
    • 9. Slides rinsed with *reaction buffer* x3
    • 10. *iView Biotinylated IgG* applied and incubated for 8 min
    • 11. Slides rinsed with *reaction buffer*
    • 14. *iView Streptavidin-Horseradish Peroxidase* applied and incubated for 8 min
    • 15. Slides rinsed with *reaction buffer*
    • 16. *iView DAB/H2O2* applied and incubated for 8 min
    • 17. Slides rinsed with *reaction buffer*
    • 18. *iView Copper* applied and incubated for 4 min
    • 19. Slides rinsed with *reaction buffer*
    • 20. Slides washed in Dawn Detergent/tap water
    • 21. Counterstain Procedure - Hematoxylin (Leica 560 MX) 30 sec - Slides washed in tap water, 1 min - Decolorized (10% Acetic Acid in 70% ethanol), 1 min - Slides washed in tap water, 1 min - Bluing (Austin Clear Ammonia), 1 min - Slides washed in tap water, 1 min
    • 22. Dehydration/coverslipping procedure: - 3 changes of 95% Ethanol, 3 min each - 3 changes of 100% Ethanol, 3 min each - 3 changes of Xylene, 5 min each - Mounted with Permount
    • 23. Imaging: Leica SCN 400F Whole Slide Scanner with Digital Image Hub and Leica Slidepath software
    Experimental Notes
    • Deviations from protocol/procedure supplied by manufacturer:
    • Step 1: Heated tissue 60°C for 30 minutes; manufacturer heats for 45 minutes.
    • Step 2: No ethanol wash was performed during deparaffinization; manufacturer includes 1 wash of 80% ethanol for 3 minutes.
    • Step 3.1: Slides were heated for 15 minutes; manufacturer provides a range of 15-20 minutes.
    • Step 3.2: Slides were cooled for 30 minutes; manufacturer cools for 20 minutes.
    • Step 4: Italicized reagents and incubation time are fixed instrument parameters.
    • Step 5: Secondary species-specific serum block not used; manufacturer blocks with 5% normal goat serum for 2 hours.
    • Step 8.1: Antibody diluted in PBS at 1:250; manufacture did not recommend diluent or dilution.
    • Step 8.2.1: Primary antibody incubated at room temperature overnight; manufacturer incubates overnight 4°C with agitation.
  • 状态
    Liquid
    浓度
    1 μg/μL
    缓冲液
    0.01M TBS( pH 7.4) with 1 % BSA, 0.02 % Proclin300 and 50 % Glycerol.
    储存液
    ProClin
    注意事项
    This product contains ProClin: a POISONOUS AND HAZARDOUS SUBSTANCE, which should be handled by trained staff only.
    储存条件
    4 °C,-20 °C
    储存方法
    Shipped at 4°C. Store at -20°C for one year. Avoid repeated freeze/thaw cycles.
    有效期
    12 months
  • Wang, Fu, Jiang, Yu, Wang, Han, Liu, Wang: "Bone marrow mononuclear cell transplantation promotes therapeutic angiogenesis via upregulation of the VEGF-VEGFR2 signaling pathway in a rat model of vascular dementia." in: Behavioural brain research, Vol. 265, pp. 171-80, (2014) (PubMed).

  • 抗原
    RAF1 (V-Raf-1 Murine Leukemia Viral Oncogene Homolog 1 (RAF1))
    别名
    c-Raf/Raf1 (RAF1 产品)
    别名
    CRAF antibody, NS5 antibody, Raf-1 antibody, c-Raf antibody, c-raf antibody, craf antibody, raf1 antibody, RAF1 antibody, 6430402F14Rik antibody, AA990557 antibody, BB129353 antibody, Craf1 antibody, D830050J10Rik antibody, v-Raf antibody, raf antibody, cRaf antibody, Raf-1 proto-oncogene, serine/threonine kinase antibody, Raf-1 proto-oncogene, serine/threonine kinase a antibody, v-raf-leukemia viral oncogene 1 antibody, Raf-1 proto-oncogene, serine/threonine kinase S homeolog antibody, RAF1 antibody, Raf1 antibody, raf1a antibody, raf1 antibody, raf1.S antibody
    背景

    Synonyms: NS5, CRAF, Raf-1, c-Raf, CMD1NN, RAF proto-oncogene serine/threonine-protein kinase, Proto-oncogene c-RAF, RAF1, RAF

    Background: 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ID
    5894
    UniProt
    P04049
    途径
    MAPK Pathway, RTK signaling, Fc-epsilon Receptor Signaling Pathway, Neurotrophin Signaling Pathway, cAMP Metabolic Process, Stem Cell Maintenance, Hepatitis C, Autophagy, Signaling of Hepatocyte Growth Factor Receptor, VEGF Signaling, BCR Signaling
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