Anti-STAT3 抗体 [EPR361] (ab109085)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR361] to STAT3
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
-
製品名
Anti-STAT3 antibody [EPR361]
STAT3 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR361] to STAT3 -
由来種
Rabbit -
アプリケーション
適用あり: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details
適用なし: IP -
種交差性
交差種: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
-
ポジティブ・コントロール
- WB: HAP1, HEK293, A431, Raji, HeLa, HaCaT, NIH/3T3, C2C12, and SH-SY5Y cell lysates. Human, mouse, and rat brain, mouse heart, and rat kidney tissue lysates. ICC/IF: HeLa and A459 cells. IHC-P: Human pancreas tissue. Flow Cyt (intra): HeLa cells.
-
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 -20°C. Stable for 12 months at -20°C. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA -
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
EPR361 -
アイソタイプ
IgG -
研究分野
関連製品
-
Alternative Versions
-
Assay kits
-
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ab109085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) |
1/300.
|
|
WB |
1/1000 - 1/10000. Predicted molecular weight: 88 kDa.
|
|
IHC-P |
1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
|
|
ICC/IF |
1/150.
|
特記事項 |
---|
Flow Cyt (Intra)
1/300. |
WB
1/1000 - 1/10000. Predicted molecular weight: 88 kDa. |
IHC-P
1/100. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
ICC/IF
1/150. |
ターゲット情報
-
機能
Signal transducer and transcription activator that mediates cellular responses to interleukins, KITLG/SCF, LEP and other growth factors. Once activated, recruits coactivators, such as NCOA1 or MED1, to the promoter region of the target gene (PubMed:17344214). May mediate cellular responses to activated FGFR1, FGFR2, FGFR3 and FGFR4. Binds to the interleukin-6 (IL-6)-responsive elements identified in the promoters of various acute-phase protein genes. Activated by IL31 through IL31RA. Involved in cell cycle regulation by inducing the expression of key genes for the progression from G1 to S phase, such as CCND1 (PubMed:17344214). Mediates the effects of LEP on melanocortin production, body energy homeostasis and lactation (By similarity). May play an apoptotic role by transctivating BIRC5 expression under LEP activation (PubMed:18242580). Cytoplasmic STAT3 represses macroautophagy by inhibiting EIF2AK2/PKR activity. -
組織特異性
Heart, brain, placenta, lung, liver, skeletal muscle, kidney and pancreas. -
関連疾患
Hyperimmunoglobulin E recurrent infection syndrome, autosomal dominant
Autoimmune disease, multisystem, infantile-onset -
配列類似性
Belongs to the transcription factor STAT family.
Contains 1 SH2 domain. -
翻訳後修飾
Tyrosine phosphorylated upon stimulation with EGF. Tyrosine phosphorylated in response to constitutively activated FGFR1, FGFR2, FGFR3 and FGFR4 (By similarity). Activated through tyrosine phosphorylation by BMX. Tyrosine phosphorylated in response to IL6, IL11, LIF, CNTF, KITLG/SCF, CSF1, EGF, PDGF, IFN-alpha, LEP and OSM. Activated KIT promotes phosphorylation on tyrosine residues and subsequent translocation to the nucleus. Phosphorylated on serine upon DNA damage, probably by ATM or ATR. Serine phosphorylation is important for the formation of stable DNA-binding STAT3 homodimers and maximal transcriptional activity. ARL2BP may participate in keeping the phosphorylated state of STAT3 within the nucleus. Upon LPS challenge, phosphorylated within the nucleus by IRAK1. Upon erythropoietin treatment, phosphorylated on Ser-727 by RPS6KA5. Phosphorylation at Tyr-705 by PTK6 or FER leads to an increase of its transcriptional activity. Dephosphorylation on tyrosine residues by PTPN2 negatively regulates IL6/interleukin-6 signaling. -
細胞内局在
Cytoplasm. Nucleus. Shuttles between the nucleus and the cytoplasm. Translocated into the nucleus upon tyrosine phosphorylation and dimerization, in response to signaling by activated FGFR1, FGFR2, FGFR3 or FGFR4. Constitutive nuclear presence is independent of tyrosine phosphorylation. Predominantly present in the cytoplasm without stimuli. Upon leukemia inhibitory factor (LIF) stimulation, accumulates in the nucleus. The complex composed of BART and ARL2 plays an important role in the nuclear translocation and retention of STAT3. Identified in a complex with LYN and PAG1. - Information by UniProt
-
参照データベース
- Entrez Gene: 6774 Human
- Entrez Gene: 20848 Mouse
- Entrez Gene: 25125 Rat
- Omim: 102582 Human
- SwissProt: P40763 Human
- SwissProt: P42227 Mouse
- SwissProt: P52631 Rat
- Unigene: 463059 Human
see all -
別名
- 1110034C02Rik antibody
- Acute Phase Response Factor antibody
- Acute-phase response factor antibody
see all
画像
-
All lanes : Anti-STAT3 antibody [EPR361] (ab109085) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : STAT3 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 88 kDa
Observed band size: 92 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab109085 observed at 92 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab109085 was shown to react with STAT3 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255436 (knockout cell lysate ab263797) was used. Wild-type HeLa and STAT3 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. ab109085 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.
-
Immunohistochemical staining of paraffin embedded human pancreas with purified ab109085 at a working dilution of 1 in 100. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
-
Immunocytochemistry/Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) labeling STAT3 with Purified ab109085 at 1/250 dilution (5 µg/ml). Cells were fixed with 4% PFA and permeabilized with 0.1% tritonX-100. ab150077 Goat anti rabbit IgG(Alexa Fluor® 488) at 1/1000 dilution was used as the secondary antibody. Nuclei were counterstained with DAPI. PBS was used instead of the primary antibody as the negative control.
-
All lanes : Anti-STAT3 antibody [EPR361] (ab109085) at 1/1000 dilution
Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : HaCaT (Human skin keratinocyte) whole cell lysate
Lane 3 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 4 : C2C12 (Mouse myoblasts cell line) whole cell lysate
Lane 5 : Human brain lysate
Lane 6 : Mouse brain lysate
Lane 7 : Rat brain lysate
Lane 8 : Rat liver lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 88 kDa
Observed band size: 92 kDa why is the actual band size different from the predicted?
Additional bands at: 32 kDa, 40 kDa. We are unsure as to the identity of these extra bands.
Exposure time: 50 secondsBlocking/Diluting buffer: 5% NFDM/TBST.
Rabbit monoclonal [EPR16891] to GAPDH (ab181602) used as loading control.
-
All lanes : Anti-STAT3 antibody [EPR361] (ab109085) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : STAT3 knockout HAP1 whole cell lysate
Lane 3 : HeLa whole cell lysate
Lane 4 : HEK293 whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 88 kDaLanes 1 - 4: Merged signal (red and green). Green - ab109085 observed at 92 kDa. Red - loading control, ab8245, observed at 37 kDa.
Ab109085 was shown to specifically react with STAT3 in wild-type cells as signal was lost in STAT3 knockout HAP1 cells. Wild-type and STAT3 knockout samples were subjected to SDS-PAGE. Ab109085 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.
-
Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling STAT3 (red) with ab109085 at a 1/300 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluorr® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.
-
All lanes : Anti-STAT3 antibody [EPR361] (ab109085) at 1/2000 dilution (purified)
Lane 1 : Mouse heart tissue lysate
Lane 2 : Rat brain tissue lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 88 kDa
Observed band size: 88 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
-
Immunofluorescence staining of A549 cells with purified ab109085 at a working dilution of 1 in 150, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 555 goat anti rabbit (ab150078), used at a dilution of 1 in 500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, purified ab109085 was used at a dilution of 1/200 followed by an Alexa Fluor® 488 goat anti-mouse antibody at a dilution of 1/500.
-
All lanes : Anti-STAT3 antibody [EPR361] (ab109085) at 1/2000 dilution (purified)
Lane 1 : A431 cell lysate
Lane 2 : Raji cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : SH-S5SY (Human neuroblastoma cell line from bone marrow) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1000 mg/ml
Predicted band size: 88 kDa
Observed band size: 88 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
-
All lanes : Anti-STAT3 antibody [EPR361] (ab109085) at 1/1000 dilution (unpurified)
Lane 1 : A431 cell lysate
Lane 2 : Raji cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : SH-SY5Y cell lysate
Lysates/proteins at 10 µg per lane.
Predicted band size: 88 kDa
プロトコール
データシートおよび資料
-
SDS download
-
Datasheet download
参考文献 (19)
ab109085 は 19 報の論文で使用されています。
- Liao H et al. B7‑H3 promotes the epithelial‑mesenchymal transition of NSCLC by targeting SIRT1 through the PI3K/AKT pathway. Mol Med Rep 25:N/A (2022). PubMed: 35029291
- Xu GY et al. Cell-Free Extracts from Human Fat Tissue with a Hyaluronan-Based Hydrogel Attenuate Inflammation in a Spinal Cord Injury Model through M2 Microglia/Microphage Polarization. Small 18:e2107838 (2022). PubMed: 35333441
- Li X et al. New bitongling (NBTL) ameliorates rheumatoid arthritis in rats through inhibiting JAK2/STAT3 signaling pathway. Eur J Histochem 65:N/A (2021). PubMed: 33634679
- Zhou Y et al. Globular adiponectin inhibits osteoblastic differentiation of vascular smooth muscle cells through the PI3K/AKT and Wnt/β-catenin pathway. J Mol Histol 52:1067-1080 (2021). PubMed: 34398360
- Chen Q et al. TRIM18-Regulated STAT3 Signaling Pathway via PTP1B Promotes Renal Epithelial-Mesenchymal Transition, Inflammation, and Fibrosis in Diabetic Kidney Disease. Front Physiol 12:709506 (2021). PubMed: 34434118