RAF1 抗体 (AA 31-130)
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- 抗原 See all RAF1 抗体
- RAF1 (V-Raf-1 Murine Leukemia Viral Oncogene Homolog 1 (RAF1))
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抗原表位
- AA 31-130
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适用
- 人
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宿主
- 兔
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克隆类型
- 多克隆
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标记
- This RAF1 antibody is un-conjugated
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应用范围
- 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
- 人
- 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
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- 应用备注
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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
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- by
- Immunohistochemistry Core, NYU Langone
- No.
- #029651
- 日期
- 2014.04.01
- 抗原
- Lot Number
- 130909
- Method validated
- Immunohistochemistry
- Positive Control
- Human colon villus epithelial cells
- 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.
- 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.
生效 #029651 (Immunohistochemistry)Validation ImagesFull Methods -
- 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
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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).
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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).
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- 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
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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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