Phospho-BRAF (T401) Recombinant Rabbit Monoclonal Antibody [JJ08-72]
Catalog# ET1701-20
Phospho-BRAF (T401) Recombinant Rabbit Monoclonal Antibody [JJ08-72]
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WB
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IF-Cell
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IP
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IHC-P
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Human
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Mouse
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Rat
概述
产品名称
Phospho-BRAF (T401) Recombinant Rabbit Monoclonal Antibody [JJ08-72]
抗体类型
Recombinant Rabbit monoclonal Antibody
免疫原
Synthetic phospho-peptide corresponding to residues surrounding Thr401 of Human BRAF aa 371-420 / 766.
种属反应性
Human, Mouse, Rat
验证应用
WB, IF-Cell, IP, IHC-P
分子量
Predicted band size: 84 kDa
阳性对照
Mouse testis tissue lysates, PC-12 cell lysates, PC-3M, MCF-7, PC-12, human testis tissue, mouse testis tissue, rat testis tissue, mouse brain tissue, rat brain tissue.
偶联
unconjugated
克隆号
JJ08-72
RRID
产品特性
形态
Liquid
存放说明
Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
存储缓冲液
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
亚型
IgG
纯化方式
Protein A affinity purified.
应用稀释度
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WB
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1:2,000
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IF-Cell
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1:100
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IP
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Use at an assay dependent concentration.
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IHC-P
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1:200
靶点
功能
BRAF is a human gene that encodes a protein called B-Raf. The gene is also referred to as proto-oncogene B-Raf and v-Raf murine sarcoma viral oncogene homolog B, while the protein is more formally known as serine/threonine-protein kinase B-Raf. B-Raf is a member of the Raf kinase family of growth signal transduction protein kinases. This protein plays a role in regulating the MAP kinase/ERKs signaling pathway, which affects cell division, differentiation, and secretion. The B-Raf protein is involved in sending signals inside cells which are involved in directing cell growth. Certain other inherited BRAF mutations cause birth defects. Drugs that treat cancers driven by BRAF mutations have been developed. Two of these drugs, vemurafenib and dabrafenib are approved by FDA for treatment of late-stage melanoma.
背景文献
1. Brown WS et al. B-Raf regulation of integrin a4 1-mediated resistance to shear stress through changes in cell spreading and cytoskeletal association in T cells. J Biol Chem 289:23141-53 (2014).
2. Wada, M. et al. The dual RAF/MEK inhibitor CH5126766/RO5126766 may be a potential therapy for RAS-mutated tumor cells. PloS one. 9: e113217(2014).
序列相似性
Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. RAF subfamily.
组织特异性
Brain and testis.
翻译后修饰
Phosphorylation at Ser-365 by SGK1 inhibits its activity.; Methylation at Arg-671 decreases stability and kinase activity.; Ubiquitinated by RNF149; which leads to proteasomal degradation. Polyubiquitinated at Lys-578 in response to EGF.
亚细胞定位
Nucleus, Cytoplasm, Cell membrane.
别名
FLJ95109 antibody
94 kDa B raf protein antibody
B raf 1 antibody
B Raf proto oncogene serine threonine protein kinase antibody
B Raf proto oncogene, serine/threonine kinase antibody
B RAF1 antibody
B-Raf proto-oncogene serine/threonine-protein kinase (p94) antibody
BRAF 1 antibody
BRAF antibody
BRAF_HUMAN antibody
展开图片
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☑ Cell treatment (CT)
Western blot analysis of Phospho-BRAF (T401) on different lysates with Rabbit anti-Phospho-BRAF (T401) antibody (ET1701-20) at 1/2,000 dilution.
Lane 1: C6 cell lysate
Lane 2: C6 starved overnight then treated with 200nM TPA for 4 hours cell lysate
Lane 3: C6 starved overnight then treated with 200nM TPA for 4 hours cell lysate, then the membrane treated with λpp for 1 hour
Lysates/proteins at 20 µg/Lane.
Predicted band size: 84 kDa
Observed band size: 84 kDa
Exposure time: 3 minutes; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1701-20) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Cell treatment (CT)
Immunohistochemical analysis of paraffin-embedded human testis tissue untreated / treated with λpp with Rabbit anti-Phospho-BRAF (T401) antibody (ET1701-20) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-20) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse testis tissue with Rabbit anti-Phospho-BRAF (T401) antibody (ET1701-20) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-20) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat testis tissue with Rabbit anti-Phospho-BRAF (T401) antibody (ET1701-20) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-20) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded mouse brain tissue with Rabbit anti-Phospho-BRAF (T401) antibody (ET1701-20) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-20) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-Phospho-BRAF (T401) antibody (ET1701-20) at 1/200 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1701-20) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. -
ICC staining of Phospho-BRAF (T401) in PC-3M cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-20, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining of Phospho-BRAF (T401) in MCF-7 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-20, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
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ICC staining of Phospho-BRAF (T401) in PC-12 cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 10% negative goat serum for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1701-20, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
请注意: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
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