Product datasheet

Anti-STAT3 antibody [EPR787Y] ab68153
リコンビナント

10 Abreviews 352 References 画像数 8
製品の概要
製品名

Anti-STAT3 antibody [EPR787Y]

製品の詳細

Rabbit monoclonal [EPR787Y] to STAT3

由来種

Rabbitアプリケーション

適用あり : Flow Cyt (Intra), ChIC/CUT&RUN-seq, ICC/IF, WB, IHC-P
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: Rat and mouse heart tissue lysates; Human, mouse, and rat brain tissue lysates; Rat kidney
tissue lysate; HeLa, HAP1, A431, HaCaT, NIH/3T3, C2C12, and Raji cell lysates. IHC-P: Humanbrain tissue. ICC/IF: HeLa cells. Flow Cyt (intra): Raji cells. ChIC/CUT&RUN seq: HepG2 cell

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
For more information see here.
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.
Avoid freeze / thaw cycle.

バッファー

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS

精製度

Protein A purified

ポリ /モノモノクローナル

クローン名

EPR787Y
1アイソタイプ

IgG

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ab68153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。
アプリケーション特記事項

Abreviews

Flow Cyt (Intra)

1/30 - 1/50.

ChIC/CUT&RUN-seq

Use at an assay dependent concentration.

ICC/IF(1)

1/200. For unpurified use at 1/140.

WB

(7)

1/1000 - 1/2000. Detects a band of approximately 75, 88 kDa(predicted molecular weight: 88 kDa).

IHC-P

(1)

1/200. Perform heat mediated antigen retrieval with Tris/EDTA
buffer pH 9.0 before commencing with IHC staining protocol.

For unpurified use at 1/140.
追加情報

Is unsuitable for IP.

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 NCOA1or MED1, to the promoter region of the target gene (PubMed:17344214). May mediate cellularresponses 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. Activatedby 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 theeffects 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 dominantAutoimmune 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 toconstitutively activated FGFR1, FGFR2, FGFR3 and FGFR4 (By similarity). Activated throughtyrosine 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 phosphorylation2

is important for the formation of stable DNA-binding STAT3 homodimers and maximaltranscriptional activity. ARL2BP may participate in keeping the phosphorylated state of STAT3
within the nucleus. Upon LPS challenge, phosphorylated within the nucleus by IRAK1. Uponerythropoietin 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 thenucleus 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.

画像ChIC/CUT&RUN was performed using a pAG-MNAse at a final
concentration of 700 ng/mL, 2.5 x 10^5 HepG2 cells (starvedovernight and treated with 100ng/ml IL-6 for 30min) and 5 µg ofab68153 [EPR787Y]. The resulting DNA was sequenced on theIllumina NovaSeq 6000 to a depth of 10 million reads. The negative
IgG control ab172730 is also shown.
Additional screenshots of mapped reads can be downloaded here.
The University of Geneva owns patents relevant to ChIC (ChromatinImmuno-Cleavage) methods.

ChIC/CUT&RUN sequencing - Anti-STAT3 antibody[EPR787Y] (ab68153)

All lanes : Anti-STAT3 antibody [EPR787Y] (ab68153) 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
Western blot - Anti-STAT3 antibody [EPR787Y]

Observed band size: 92 kDa(ab68153)3

Lanes 1- 2: Merged signal (red and green). Green - ab68153
observed at 92 kDa. Red - Anti-GAPDH antibody [6C5] - LoadingControl (ab8245) observed at 37 kDa.
ab68153 was shown to react with STAT3 in wild-type HeLa cells inwestern 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 toSDS-PAGE. Membrane was blocked for 1 hour at room
temperature in 0.1% TBST with 3% non-fat dried milk. ab68153 andAnti-GAPDH antibody [6C5] - Loading Control (ab8245) overnightat 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-MouseIgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary
antibodies at 1 in 20000 dilution for 1 hour at room temperature
before imaging.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embeddedsections) analysis of human brain tissue sections labelling STAT3
with purified ab68153 at 1/200. Heat mediated antigen retrieval
was performed using Tris/EDTA buffer pH 9. A prediluted HRPpolymer conjugated anti-rabbit IgG was used as the secondary
antibody. Counterstained with Hematoxylin.

Immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections) - Anti-STAT3 antibody[EPR787Y] (ab68153)

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All lanes : Anti-STAT3 antibody [EPR787Y] (ab68153) 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 myoblast cell line) whole cell lysate
Lane 5 : Human brain lysate
Lane 6 : Mouse brain lysate
Lane 7 : Rat brain lysate
Western blot - Anti-STAT3 antibody [EPR787Y]

Lane 8 : Rat kidney lysate(ab68153)

Lysates/proteins at 20 µg per lane.

SecondaryAll lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000
dilution

Predicted band size: 88 kDa
Observed band size: 92 kDa

Exposure time: 90 seconds

Blocking/Diluting buffer: 5% NFDM/TBST.
Rabbit monoclonal [EPR16891] to GAPDH (ab181602) used asloading control.

5All lanes : Anti-STAT3 antibody [EPR787Y] (ab68153) at 1/500
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 kDa
Western blot - Anti-STAT3 antibody [EPR787Y](ab68153)

Lanes 1 - 4: Merged signal (red and green). Green - ab68153
observed at 92 kDa. Red - loading control, ab8245, observed at 37kDa.
Ab68153 detected the expected band for STAT3 in wild-type cells
along with additional cross-reactive bands. The band was not seen
in STAT3 knockout HAP1 cells. Wild-type and STAT3 knockout
samples were subjected to SDS-PAGE. Ab68153 and ab8245(Mouse anti GAPDH loading control) were incubated overnight at4°C at 1/500 dilution and 1/10000 dilution respectively. Blots weredeveloped 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.

Immunocytochemistry/Immunofluorescence analysis of HeLa cells
labelling STAT3 (green) with purified ab68153 at 1/200. Cells were
fixed with 4% paraformaldehyde. An Alexa Fluor® 488-conjugatedgoat anti-rabbit IgG (1/200) was used as the secondary antibody.
DAPI (blue) was used as the nuclear counterstain.

Immunocytochemistry/ Immunofluorescence - AntiSTAT3 antibody [EPR787Y] (ab68153)

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ab68153 stainingSTAT3in the human cell line HeLa (human cervixadenocarcinoma) by intracellular flow cytometry. Cells were fixedwith 4% paraformaldehyde, permeabilized with 90% methanol andthe sample was incubated with the primary antibody at a dilution of
1/30. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of
1/2000 was used as the secondary antibody.
Isoytype control: Rabbit monoclonal IgG (Black)
Unlabelled control: Cell without incubation with primary antibody andsecondary antibody (Blue)

Flow Cytometry (Intracellular) - Anti-STAT3 antibody[EPR787Y] (ab68153)

Anti-STAT3 antibody [EPR787Y] (ab68153)

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