Anti-BRAF (mutated V600E) 抗体 [VE1] (ab228461)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Mouse monoclonal [VE1] to BRAF (mutated V600E)
- Suitable for: IHC-P, WB
- Reacts with: Human
製品の概要
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製品名
Anti-BRAF (mutated V600E) antibody [VE1]
BRAF 一次抗体 製品一覧 -
製品の詳細
Mouse monoclonal [VE1] to BRAF (mutated V600E) -
由来種
Mouse -
特異性
The VE1 monoclonal is a sensitive antibody that detects mutated, constitutively active BRAF protein where glutamic acid is present at codon 600 instead of valine (V600E) (PubMed IDs: 21638088, 23657789).
Please be aware that non-specific nuclear staining has been reported with this antibody (PubMed IDs: 23763264, 23589031, 24838325).
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アプリケーション
適用あり: IHC-P, WBmore details -
種交差性
交差種: Human
交差が予測される動物種: Mouse, Chicken -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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ポジティブ・コントロール
- WB: A375 cell lysate. IHC-P: A375, Caco-2 and Human melanoma tissue with B-RAF V600E mutation.
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特記事項
The V600E activating mutation in BRAF is found in several cancer types including: ~65% of pleomorphic xanthoastrocytomas, ~52% of microsatellite-unstable colon cancer tumors, ~50% of melanomas, ~35% of papillary thyroid carcinomas and ~5% of microsatellite-stable colon cancers (PubMed IDs: 21274720, 24508103, 18682506, 16024606).
The majority (>90%) of BRAF mutant cancers harbor the V600E mutation. The mutation leads to activation of the MAPK signaling pathway that increases cell invasion and reduces apoptosis. It also leads to reduced expression of melanocyte differentiation antigens and subsequent immune evasion (PubMed IDs: 21638088, 20551059).
The VE1 monoclonal antibody is manufactured by Abcam. If you require a particular buffer formulation or a particular conjugate for your experiments, please see Custom formulation and conjugation services
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@abcam.com.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
バッファー
pH: 7.50
Preservative: 0.1% Sodium azide
Constituents: Tris, 0.3% Carrier protein -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
VE1 -
アイソタイプ
IgG2b -
軽鎖の種類
kappa -
研究分野
関連製品
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Compatible Secondaries
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Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab228461の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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IHC-P | (1) |
1/100.
Perform heat mediated epitope retrieval with citrate buffer pH6 before commencing with IHC staining protocol |
WB | (2) |
1/1000. Predicted molecular weight: 84 kDa.
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特記事項 |
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IHC-P
1/100. Perform heat mediated epitope retrieval with citrate buffer pH6 before commencing with IHC staining protocol |
WB
1/1000. Predicted molecular weight: 84 kDa. |
ターゲット情報
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機能
Involved in the transduction of mitogenic signals from the cell membrane to the nucleus. May play a role in the postsynaptic responses of hippocampal neuron. -
組織特異性
Brain and testis. -
関連疾患
Note=Defects in BRAF are found in a wide range of cancers.
Defects in BRAF may be a cause of colorectal cancer (CRC) [MIM:114500].
Defects in BRAF are involved in lung cancer (LNCR) [MIM:211980].
Defects in BRAF are involved in non-Hodgkin lymphoma (NHL) [MIM:605027]. NHL is a cancer that starts in cells of the lymph system, which is part of the body's immune system. NHLs can occur at any age and are often marked by enlarged lymph nodes, fever and weight loss.
Defects in BRAF are a cause of cardiofaciocutaneous syndrome (CFC syndrome) [MIM:115150]; also known as cardio-facio-cutaneous syndrome. CFC syndrome is characterized by a distinctive facial appearance, heart defects and mental retardation. Heart defects include pulmonic stenosis, atrial septal defects and hypertrophic cardiomyopathy. Some affected individuals present with ectodermal abnormalities such as sparse, friable hair, hyperkeratotic skin lesions and a generalized ichthyosis-like condition. Typical facial features are similar to Noonan syndrome. They include high forehead with bitemporal constriction, hypoplastic supraorbital ridges, downslanting palpebral fissures, a depressed nasal bridge, and posteriorly angulated ears with prominent helices. The inheritance of CFC syndrome is autosomal dominant.
Defects in BRAF are the cause of Noonan syndrome type 7 (NS7) [MIM:613706]. Noonan syndrome is a disorder characterized by facial dysmorphic features such as hypertelorism, a downward eyeslant and low-set posteriorly rotated ears. Other features can include short stature, a short neck with webbing or redundancy of skin, cardiac anomalies, deafness, motor delay and variable intellectual deficits.
Defects in BRAF are the cause of LEOPARD syndrome type 3 (LEOPARD3) [MIM:613707]. LEOPARD3 is a disorder characterized by lentigines, electrocardiographic conduction abnormalities, ocular hypertelorism, pulmonic stenosis, abnormalities of genitalia, retardation of growth, and sensorineural deafness.
Note=A chromosomal aberration involving BRAF is found in pilocytic astrocytomas. A tandem duplication of 2 Mb at 7q34 leads to the expression of a KIAA1549-BRAF fusion protein with a constitutive kinase activity and inducing cell transformation. -
配列類似性
Belongs to the protein kinase superfamily. TKL Ser/Thr protein kinase family. RAF subfamily.
Contains 1 phorbol-ester/DAG-type zinc finger.
Contains 1 protein kinase domain.
Contains 1 RBD (Ras-binding) domain. -
細胞内局在
Nucleus. Cytoplasm. Cell membrane. Colocalizes with RGS14 and RAF1 in both the cytoplasm and membranes. - Information by UniProt
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参照データベース
- Entrez Gene: 673 Human
- Entrez Gene: 109880 Mouse
- Omim: 164757 Human
- SwissProt: Q04982 Chicken
- SwissProt: P15056 Human
- SwissProt: P28028 Mouse
- Unigene: 550061 Human
- Unigene: 245513 Mouse
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別名
- FLJ95109 antibody
- 94 kDa B raf protein antibody
- B raf 1 antibody
see all
画像
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Immunohistochemical analysis of formalin fixed paraffin embedded human melanoma labelling BRAF (mutated V600E) with ab228461 at 1/600 dilution. The immunostaining was performed on a Ventana DISCOVERY ULTRA (Roche Tissue Diagnostics) instrument with an OptiView DAB IHC Detection Kit. Heat mediated antigen retrieval was conducted for 32 mins at 100°C with ULTRA cell conditioning solution (CC1, pH 8.5). ab228461 anti-BRAF (mutated V600E) antibody [VE1] was incubated at 37°C for 16 mins. Sections were counterstained with Hematoxylin II. Image inset shows absence of staining in secondary antibody only control.
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Immunohistochemical analysis of formalin-fixed paraffin-embedded human melanoma tissue (carrying mutant BRAF V600E) labelling BRAF (mutated V600E) with ab228461 at 1/100 dilution. The section was pre-treated using heat-mediated antigen retrieval method with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab228461, 1/100 dilution, at room temperature and detected using a Leica BOND™ Polymer refine kit.
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All lanes : Anti-BRAF (mutated V600E) antibody [VE1] (ab228461) at 1/1000 dilution
Lane 1 : HCT 116 cell lysate (wildtype BRAF)
Lane 2 : SW480 cell lysate (wildtype BRAF)
Lane 3 : Caco-2 cell lysate (wildtype BRAF)
Lane 4 : HT-29 cell lysate (mutant BRAF V600E)
Lane 5 : A375 cell lysate (mutant BRAF V600E)
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 84 kDa
Observed band size: 85 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-BRAF (mutated V600E) antibody [VE1] staining at 1/1000 dilution, shown in green; Rabbit Anti-GAPDH antibody [EPR16891] (ab181602) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab228461 was shown to bind specifically to mutant BRAF V600E. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed (ab216777) at 1/20000 dilution.
This image was generated in-house using a previous batch, manufactured using hybridoma production method.
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IHC image of BRAF (mutated V600E) staining in a section of formalin-fixed paraffin-embedded A375 cell line (carrying mutant BRAF V600E) performed on a Leica BONDTM system using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab228461, 5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
This image was generated in-house using a previous batch, manufactured using hybridoma production method.
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Negative control cell line image: IHC image of BRAF (mutated V600E) staining in a section of formalin-fixed paraffin-embedded Caco-2 cell line performed on a Leica BONDTM system using the standard protocol. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab228461, 5ug/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody.
This image was generated in-house using a previous batch, manufactured using hybridoma production method.
プロトコール
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (110)
ab228461 は 110 報の論文で使用されています。
- Rangel-Pozzo A et al. Three-dimensional telomere profiles in papillary thyroid cancer variants: A pilot study. Bosn J Basic Med Sci 22:481-487 (2022). PubMed: 34882527
- Oh KY et al. BRAF V600E and previously unidentified KRAS G12C mutations in odontogenic tumors may affect MAPK activation differently depending on tumor type. Genes Chromosomes Cancer 61:481-490 (2022). PubMed: 35353428
- Shaalan AAM et al. Prognostic Value of LINC-ROR (rs1942347) Variant in Patients with Colon Cancer Harboring BRAF Mutation: A Propensity Score-Matched Analysis. Biomolecules 12:N/A (2022). PubMed: 35454158
- Zurnadzhy L et al. Clinicopathological Implications of the BRAFV600E Mutation in Papillary Thyroid Carcinoma of Ukrainian Patients Exposed to the Chernobyl Radiation in Childhood: A Study for 30 Years After the Accident. Front Med (Lausanne) 9:882727 (2022). PubMed: 35665338
- Bogdanova T et al. The high degree of similarity in histopathological and clinical characteristics between radiogenic and sporadic papillary thyroid microcarcinomas in young patients. Front Endocrinol (Lausanne) 13:970682 (2022). PubMed: 36060986