Product datasheet

Anti-c-Jun antibody [EP693Y] ab40766
リコンビナント

1 Abreviews 44 References 画像数 11
製品の概要
製品名

Anti-c-Jun antibody [EP693Y]

製品の詳細

Rabbit monoclonal [EP693Y] to c-Jun

由来種

Rabbit

特異性

PBS only lot tested.

アプリケーション

適用あり : Flow Cyt (Intra), ICC/IF, WB, IHC-P, 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: HEK-293, MOJ/3T3 and PC-12 cell lysates;. IHC-P: Rat liver, mouse cerebrum and humancervix carcinoma tissues. ICC/IF: NIH/3T3 and HeLa cells. ICC/IF KO: HEK293 cells (HEK293JUN KO cells used as a negative cell line). Flow Cyt (intra): HEK-293 cells IP: NIH/3T3 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
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.
Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react withthese species. Please contact us for more information.

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: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol (glycerin, glycerine), 59% PBS, 0.05% BSA1精製度Protein A purified

ポリ /モノモノクローナル

クローン名EP693Y

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ab40766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)

1/20.

ICC/IF

1/50.
Signal can be observed in cells fixed with either methanol orparaformaldehyde.

WB

1/1000 - 1/5000. Detects a band of approximately 39 kDa(predicted molecular weight: 39 kDa).

IHC-P

(1)

IP

1/500. Perform heat mediated antigen retrieval with citrate buffer
pH 6 before commencing with IHC staining protocol.
1/20.

ターゲット情報機能

Transcription factor that recognizes and binds to the enhancer heptamer motif 5'-TGA[CG]TCA-3'.
Promotes activity of NR5A1 when phosphorylated by HIPK3 leading to increased steroidogenic
gene expression upon cAMP signaling pathway stimulation. Involved in activated KRAS-mediatedtranscriptional activation of USP28 in colorectal cancer (CRC) cells (PubMed:24623306). Binds
to the USP28 promoter in colorectal cancer (CRC) cells (PubMed:24623306).

配列類似性

Belongs to the bZIP family. Jun subfamily.
Contains 1 bZIP (basic-leucine zipper) domain.

翻訳後修飾

Ubiquitinated by the SCF(FBXW7), leading to its degradation. Ubiquitination takes placefollowing phosphorylation, that promotes interaction with FBXW7.
Phosphorylated by CaMK4 and PRKDC; phosphorylation enhances the transcriptional activity.
Phosphorylated by HIPK3. Phosphorylated by DYRK2 at Ser-243; this primes the protein forsubsequent phosphorylation by GSK3B at Thr-239. Phosphorylated at Thr-239, Ser-243 and Ser249 by GSK3B; phosphorylation reduces its ability to bind DNA. Phosphorylated by PAK2 at Thr2, Thr-8, Thr-89, Thr-93 and Thr-286 thereby promoting JUN-mediated cell proliferation andtransformation. Phosphorylated by PLK3 following hypoxia or UV irradiation, leading to increase
DNA-binding activity.
Acetylated at Lys-271 by EP300.

細胞内局在Nucleus.

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画像

Purified ab40766 at 1/20 dilution (1µg) immunoprecipitating c-Jun
in NIH/3T3 whole cell lysate.
Lane 1 (input): NIH/3T3 (Mouse embryonic fibroblast) whole celllysate 10µg
Lane 2 (+): ab40766 + NIH/3T3 whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab40766
in NIH/3T3 whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/1000
dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Immunoprecipitation - Anti-c-Jun antibody [EP693Y]

Observed band size: 39 kDa(ab40766)

All lanes : Anti-c-Jun antibody [EP693Y] (ab40766) at 1/1000
dilution (Purified)

Lane 1 : HEK-293 (Human embryonic kidney epithelial cell) wholecell lysate

Lane 2 : NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma ) whole celllysate
Lysates/proteins at 15 µg per lane.

Western blot - Anti-c-Jun antibody [EP693Y]

Secondary(ab40766)

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

Predicted band size: 39 kDa
Observed band size: 39 kDa

Blocking Buffer and concentration: 5% NFDM/TBST3

Immunocytochemistry analysis of NIH/3T3 (Mouse embryonicfibroblast) cells labeling c-Jun with Purified ab40766 at 1:50 dilution(4.06 ?g/ml). Cells were fixed in 4% Paraformaldehyde andpermeabilized with 0.1% tritonX-100. Cells were counterstainedwith Ab195889 Anti-alpha Tubulin antibody [DM1A] - MicrotubuleMarker (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 thesecondary antibody only control.

Immunocytochemistry/ Immunofluorescence - Antic-Jun antibody [EP693Y] (ab40766)

Intracellular Flow Cytometry analysis of HEK-293 (Humanembryonic kidney epithelial cell) cells labeling c-Jun with Purified
ab40766 at 1/20 dilution (10µg/ml) (Red). Cells were fixed with 4%
Paraformaldehyde and permeabilised with 90% Methanol. A Goat
anti rabbit IgG (Alexa Fluorr® 488, ab150077) secondary antibody
was used at 1/2000. Isotype control - Rabbit monoclonal IgG(Black). Unlabeled control - Cell without incubation with primaryantibody and secondary antibody (Blue).

Flow Cytometry (Intracellular) - Anti-c-Jun antibody[EP693Y] (ab40766)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embeddedsections) analysis of rat liver tissue sections labeling c-Jun withPurified ab40766 at 1:500 dilution (0.41 µg/ml). Heat mediatedantigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH9.0). Rabbit specific IHC polymer detection kit HRP/DAB(ab209101) was used as the secondary antibody. Negative control:
PBS instead of the primary antibody. Hematoxylin was used as acounterstain. The immunostaining was performed on a Leica
Biosystems BOND® RX instrument.

Immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections) - Anti-c-Jun antibody [EP693Y](ab40766)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embeddedsections) analysis of mouse cerebrum tissue sections labeling cJun with Purified ab40766 at 1:500 dilution (0.41 µg/ml). Heat
mediated antigen retrieval using Bond™ Epitope Retrieval Solution
2 (pH 9.0). Rabbit specific IHC polymer detection kit HRP/DAB(ab209101) was used as the secondary antibody. Negative control:PBS instead of the primary antibody. Hematoxylin was used as acounterstain. The immunostaining was performed on a Leica
Biosystems BOND® RX instrument.

Immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections) - Anti-c-Jun antibody [EP693Y](ab40766)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embeddedsections) analysis of human cervix carcinoma tissue sections
labeling c-Jun with Purified ab40766 at 1:500 dilution (0.41 µg/ml).
Heat mediated antigen retrieval using Bond™ Epitope Retrieval
Solution 2 (pH 9.0). Rabbit specific IHC polymer detection kit
HRP/DAB (ab209101) was used as the secondary antibody.
Negative control: PBS instead of the primary antibody. Hematoxylin
was used as a counterstain. The immunostaining was performed ona Leica Biosystems BOND® RX instrument.

Immunohistochemistry (Formalin/PFA-fixed paraffinembedded sections) - Anti-c-Jun antibody [EP693Y](ab40766)

ab40766 staining c-Jun in wild-type HEK293 cells (top panel) andc-Jun knockout HEK293 cells (bottom panel). The cells were fixedwith 100% methanol (5 min), permeabilized with 0.1% Triton X-100
for 5 minutes and then blocked with 1% BSA/10% normal goat
serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were thenincubated with ab40766 at 1/250 dilution and ab7291 (Mousemonoclonal to alpha Tubulin) at 1/1000 dilution overnight at +4°C,
followed by a further incubation at room temperature for 1h with agoat secondary antibody to rabbit IgG (Alexa Fluor® 488)
(ab150081) at 2 μg/ml (shown in green) and a goat secondary
antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml
Immunocytochemistry/ Immunofluorescence - Anti-(shown in pseudo color red). Nuclear DNA was labelled in blue withc-Jun antibody [EP693Y] (ab40766)

DAPI.
Image was taken with a high-content analysis system (Perkin Elmer,Operetta CLS™).

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All lanes : Anti-c-Jun antibody [EP693Y] (ab40766) at 1/1000
dilution

Lane 1 : Wild-type HEK-293 (Human epithelial cell line fromembryonic kidney) whole cell lysate

Lane 2 : Jun knockout HEK-293 (Human epithelial cell line fromembryonic kidney) whole cell lysate
Lysates/proteins at 40 µg per lane.

Predicted band size: 39 kDa
Western blot - Anti-c-Jun antibody [EP693Y](ab40766)

Lanes 1 - 2: Merged signal (red and green). Green - ab40766
observed at 35 kDa. Red - loading control, ab18058, observed at
130 kDa.
ab40766 was shown to specifically react with Jun in wild-type HEK293 cells as signal was lost in Jun knockout cells. Wild-type and Junknockout samples were subjected to SDS-PAGE. The membrane
was blocked with 3% Milk. Ab40766 and ab18058 (Mouse antiVinculin loading control) were incubated overnight at 4°C at 1/1000
dilution and 1/10000 dilution respectively. Blots were developedwith 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.

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Immunofluorescent staining of HeLa cells
ab40766 at 1/100 dilution

Immunocytochemistry/ Immunofluorescence - Antic-Jun antibody [EP693Y] (ab40766)

Anti-c-Jun antibody [EP693Y] (ab40766)

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