Anti-ERK5 抗体 [EP791Y] (ab40809)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EP791Y] to ERK5
- Suitable for: Flow Cyt (Intra), WB, IP, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
-
製品名
Anti-ERK5 antibody [EP791Y]
ERK5 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EP791Y] to ERK5 -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), WB, IP, ICC/IFmore details -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human ERK5 aa 800-900 (C terminal). The exact sequence is proprietary.
-
ポジティブ・コントロール
- WB: HeLa, HAP1, NIH/3T3 and PC-12 cell lysates. Flow Cyt (intra): HeLa and A549 cells. ICC: HeLa cells. IP: HeLa cell lysate.
-
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.
製品の特性
-
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: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 0.05% BSA, 59% PBS -
Concentration information loading...
-
精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EP791Y -
アイソタイプ
IgG -
研究分野
関連製品
-
Alternative Versions
-
Compatible Secondaries
-
Isotype control
-
KO cell lines
-
KO cell lysates
-
Positive Controls
-
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ab40809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 |
---|---|---|
Flow Cyt (Intra) |
1/50.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/100 dilution. |
|
WB | (1) |
1/10000. Detects a band of approximately 115 kDa (predicted molecular weight: 89 kDa).
For unpurified use at 1/1000 - 1/5000 dilution. |
IP |
1/30.
For unpurified use at 1/50 dilution. |
|
ICC/IF |
1/100.
For unpurified use at 1/250 - 1/500 dilution. |
特記事項 |
---|
Flow Cyt (Intra)
1/50. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/100 dilution. |
WB
1/10000. Detects a band of approximately 115 kDa (predicted molecular weight: 89 kDa). For unpurified use at 1/1000 - 1/5000 dilution. |
IP
1/30. For unpurified use at 1/50 dilution. |
ICC/IF
1/100. For unpurified use at 1/250 - 1/500 dilution. |
ターゲット情報
-
機能
Plays a role in various cellular processes such as proliferation, differentiation and cell survival. The upstream activator of MAPK7 is the MAPK kinase MAP2K5. Upon activation, it translocates to the nucleus and phosphorylates various downstream targets including MEF2C. EGF activates MAPK7 through a Ras-independent and MAP2K5-dependent pathway. May have a role in muscle cell differentiation. May be important for endothelial function and maintenance of blood vessel integrity. MAP2K5 and MAPK7 interact specifically with one another and not with MEK1/ERK1 or MEK2/ERK2 pathways. -
組織特異性
Expressed in many adult tissues. Abundant in heart, placenta, lung, kidney and skeletal muscle. Not detectable in liver. -
配列類似性
Belongs to the protein kinase superfamily. CMGC Ser/Thr protein kinase family. MAP kinase subfamily.
Contains 1 protein kinase domain. -
ドメイン
The second proline-rich region may interact with actin targeting the kinase to a specific location in the cell.
The TXY motif contains the threonine and tyrosine residues whose phosphorylation activates the MAP kinases. -
翻訳後修飾
Dually phosphorylated on Thr-219 and Tyr-221, which activates the enzyme (By similarity). Autophosphorylated in vitro on threonine and tyrosine residues when the C-terminal part of the kinase, which could have a regulatory role, is absent. -
細胞内局在
Cytoplasm. Nucleus. Translocates to the nucleus upon activation. - Information by UniProt
-
参照データベース
- Entrez Gene: 5598 Human
- Entrez Gene: 23939 Mouse
- Entrez Gene: 114509 Rat
- Omim: 602521 Human
- SwissProt: Q13164 Human
- SwissProt: Q9WVS8 Mouse
- SwissProt: P0C865 Rat
- Unigene: 150136 Human
see all -
別名
- Big MAP kinase 1 antibody
- BMK 1 antibody
- BMK 1 kinase antibody
see all
画像
-
All lanes : Anti-ERK5 antibody [EP791Y] (ab40809) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : MAPK7 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 89 kDa
Observed band size: 115 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab40809 observed at 115 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab40809 was shown to react with ERK5 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265508 (knockout cell lysate ab258042) was used. Wild-type HeLa and MAPK7 knockout HeLa 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. ab40809 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 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.
-
Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling ERK5 with Purified ab40809 at 1:100 (4.8 µg/ml). Cells were fixed in 4% Paraformaldehyde 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 nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
-
Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling ERK5 with purified ab40809 at 1/50 dilution (10 ug/ml) (red). Cells were fixed with 80% methanol. A Goat anti rabbit IgG (Alexa Fluorr®488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
-
All lanes : Anti-ERK5 antibody [EP791Y] (ab40809) at 1/2000 dilution (Purified)
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates with 5% NFDM/TBST
Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates with 5% NFDM/TBST
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates with 5% NFDM/TBST
Lysates/proteins at 15 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Predicted band size: 89 kDa
Observed band size: 115 kDa why is the actual band size different from the predicted? -
Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
Lane 2: MAPK7 (ERK5) knockout HAP1 whole cell lysate (20 µg)
Lane 3: HeLa whole cell lysate (20 µg)Lanes 1 - 3: Merged signal (red and green). Green - ab40809 observed at 88 kDa. Red - loading control, ab9484, observed at 37 kDa.
Unpurified ab40809 was shown to specifically react with ERK5 in wild-type HAP1 cells as signal was lost in MAPK7 (ERK5) knockout cells. Wild-type and MAPK7 (ERK5) knockout samples were subjected to SDS-PAGE. Unpurified ab40809 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
-
Anti-ERK5 antibody [EP791Y] (ab40809) at 1/5000 dilution (unpurified) + Hela cell lysate at 10 µg
Predicted band size: 89 kDa
Observed band size: 115 kDa why is the actual band size different from the predicted?
The predicted weight of 89 kDa is for the precursor version of human ERK5 protein. However, ab40809 detects endogenous levels of total Erk5 protein which appears around 115 kDa in SDS PAGE -
Overlay histogram showing A549 cells stained with unpurified ab40809 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab40809, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
-
ab40809 (purified) at 1:30 dilution (2ug) immunoprecipitating ERK5 in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10ug
Lane 2 (+): ab40809 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab40809 in HeLa whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
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
-
SDS download
-
Datasheet download
参考文献 (14)
ab40809 は 14 報の論文で使用されています。
- Li J et al. Ulinastatin promotes macrophage efferocytosis and ameliorates lung inflammation via the ERK5/Mer signaling pathway. FEBS Open Bio 12:1498-1508 (2022). PubMed: 35778889
- Zhang Q et al. Isoflurane post-conditioning contributes to anti-apoptotic effect after cerebral ischaemia in rats through the ERK5/MEF2D signaling pathway. J Cell Mol Med 25:3803-3815 (2021). PubMed: 33621420
- Cilenti F et al. A PGE2-MEF2A axis enables context-dependent control of inflammatory gene expression. Immunity 54:1665-1682.e14 (2021). PubMed: 34129840
- Chen R et al. Cx43 and AKAP95 regulate G1/S conversion by competitively binding to cyclin E1/E2 in lung cancer cells. Thorac Cancer 11:1594-1602 (2020). PubMed: 32338437
- Sun Y et al. Liraglutide Promotes Osteoblastic Differentiation in MC3T3-E1 Cells by ERK5 Pathway. Int J Endocrinol 2020:8821077 (2020). PubMed: 33488706