Anti-MLK3 抗体 [EP1460Y] (ab51068)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP1460Y] to MLK3
- Suitable for: WB, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
製品の概要
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製品名
Anti-MLK3 antibody [EP1460Y]
MLK3 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP1460Y] to MLK3 -
由来種
Rabbit -
アプリケーション
適用あり: WB, ICC/IFmore details
適用なし: Flow Cyt -
種交差性
交差種: Human -
免疫原
Synthetic peptide within Human MLK3 aa 800 to the C-terminus (C terminal). The exact sequence is proprietary.
Database link: Q16584 -
ポジティブ・コントロール
- WB: HepG2, A549, HAP1, A431 and HeLa cell lysates. ICC/IF: HeLa cells.
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特記事項
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.
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: 7.20
Preservative: 0.05% Sodium azide
Constituents: 0.01% BSA, 40% Glycerol (glycerin, glycerine), 59% PBS -
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
EP1460Y -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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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ab51068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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WB |
1/1000. Detects a band of approximately 100 kDa (predicted molecular weight: 93 kDa).
For unpurified use at 1/5000. |
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ICC/IF |
1/50.
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特記事項 |
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WB
1/1000. Detects a band of approximately 100 kDa (predicted molecular weight: 93 kDa). For unpurified use at 1/5000. |
ICC/IF
1/50. |
ターゲット情報
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機能
Activates the JUN N-terminal pathway. Required for serum-stimulated cell proliferation and for mitogen and cytokine activation of MAPK14 (p38), MAPK3 (ERK) and MAPK8 (JNK1). Plays a role in mitogen-stimulated phosphorylation and activation of BRAF, but does not phosphorylate BRAF directly. Influences microtubule organization during the cell cycle. -
組織特異性
Expressed in a wide variety of normal and neoplastic tissues including fetal lung, liver, heart and kidney, and adult lung, liver, heart, kidney, placenta, skeletal muscle, pancreas and brain. -
配列類似性
Belongs to the protein kinase superfamily. STE Ser/Thr protein kinase family. MAP kinase kinase kinase subfamily.
Contains 1 protein kinase domain.
Contains 1 SH3 domain. -
翻訳後修飾
Autophosphorylation on serine and threonine residues within the activation loop plays a role in enzyme activation. Thr-277 is likely to be the main autophosphorylation site. Phosphorylation of Ser-555 and Ser-556 is induced by CDC42. -
細胞内局在
Cytoplasm > cytoskeleton > centrosome. Location is cell cycle dependent. - Information by UniProt
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参照データベース
- Entrez Gene: 4296 Human
- Omim: 600050 Human
- SwissProt: Q16584 Human
- Unigene: 502872 Human
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別名
- 2610017K16Rik antibody
- EC 2.7.11.25 antibody
- M3K11_HUMAN antibody
see all
画像
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All lanes : Anti-MLK3 antibody [EP1460Y] (ab51068) at 1/1000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate
Lane 3 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 93 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
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Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling MLK3 with Purified ab51068 at 1:50 dilution (3.52 µg/ml). Cells were fixed in 100% Methanol and permeabilized with 0.1% tritonX-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor® 488,ab150077) was used as the secondary antibody at 1:1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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All lanes : Anti-MLK3 antibody [EP1460Y] (ab51068) at 1/5000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : MAP3K11 knockout A549 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 93 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab51068 observed at 105 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab51068 was shown to react with MLK3 in wild-type A549 cells in western blot. Loss of signal was observed when knockout cell line ab267169 (knockout cell lysate ab257519) was used. Wild-type A549 and MAP3K11 knockout A549 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab51068 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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All lanes : Anti-MLK3 antibody [EP1460Y] (ab51068) at 1/5000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : MAP3K11 knockout A549 cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 93 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab51068 observed at 105 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab51068 was shown to react with MLK3 in wild-type A549 cells in western blot. Loss of signal was observed when knockout cell line ab267168 (knockout cell lysate ab257518) was used. Wild-type A549 and MAP3K11 knockout A549 cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab51068 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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Lane 1: Wild-type HAP1 cell lysate (20 µg)
Lane 2: MLK3 knockout HAP1 cell lysate (20 µg)
Lane 3: A431 cell lysate (20 µg)
Lane 4: HeLa cell lysate (20 µg)
Lanes 1 - 4: Merged signal (red and green). Green - ab51068 observed at 105 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab51068 was shown to specifically react with MLK3 when MLK3 knockout samples were used. Wild-type and MLK3 knockout samples were subjected to SDS-PAGE. ab51068 and ab8245 (loading control to GAPDH) were diluted 1/5000 and 1/1000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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 (13)
ab51068 は 13 報の論文で使用されています。
- Zhang X et al. Emodin protects H9c2 cells from hypoxia-induced injury by up-regulating miR-138 expression. Braz J Med Biol Res 52:e7994 (2019). PubMed: 30810622
- Calamaras TD et al. Mixed lineage kinase-3 prevents cardiac dysfunction and structural remodeling with pressure overload. Am J Physiol Heart Circ Physiol 316:H145-H159 (2019). PubMed: 30362822
- Xu X et al. miR-125a-5p inhibits tumorigenesis in hepatocellular carcinoma. Aging (Albany NY) 11:7639-7662 (2019). PubMed: 31527306
- Zhao H et al. Estrogen-Related Receptor ? Induces Angiogenesis and Extracellular Matrix Degradation of Temporomandibular Joint Osteoarthritis in Rats. Front Pharmacol 10:1290 (2019). PubMed: 31780931
- Lan T et al. Long non-coding RNA small nucleolar RNA host gene 12 (SNHG12) promotes tumorigenesis and metastasis by targeting miR-199a/b-5p in hepatocellular carcinoma. J Exp Clin Cancer Res 36:11 (2017). PubMed: 28073380