Anti-CEBP Beta 抗体 [E299] - C-terminal (ab32358)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [E299] to CEBP Beta - C-terminal
- Suitable for: WB, ICC/IF, IP, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
製品の概要
-
製品名
Anti-CEBP Beta antibody [E299] - C-terminal
CEBP Beta 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [E299] to CEBP Beta - C-terminal -
由来種
Rabbit -
特異性
This antibody is specific for the three CEBPB isoforms (LAP*, LAP and LIP). According to BLAST analysis, the antibody could cross-react with CEBP epsilon (32, 27 and 14kDa, 82% homology) and CEBP alpha (42kDa, 30kDa, 73% homology) in human, mouse and rat. Please be aware that this has not been confirmed experimentally. However, this could explain the background that could possibly be obtained in WB with this antibody. Please contact our Scientific Support if you have any questions.
-
アプリケーション
適用あり: WB, ICC/IF, IP, Flow Cyt (Intra)more details -
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.
(Peptide available asab274870) -
ポジティブ・コントロール
- WB: HeLa, Jurkat, PC-12, NIH/3T3 and MCF7 cell lysate. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt (intra): HeLa 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
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. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 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
E299 -
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
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ab32358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 |
---|---|---|
WB | (3) |
1/1000. Predicted molecular weight: 36 kDa.
|
ICC/IF |
1/500.
For unpurified use at 1/50 - 1/100
|
|
IP |
1/30.
For unpurified use at 1/30 |
|
Flow Cyt (Intra) |
1/600.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/1000 |
特記事項 |
---|
WB
1/1000. Predicted molecular weight: 36 kDa. |
ICC/IF
1/500. For unpurified use at 1/50 - 1/100
|
IP
1/30. For unpurified use at 1/30 |
Flow Cyt (Intra)
1/600. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/1000 |
ターゲット情報
-
機能
Important transcriptional activator in the regulation of genes involved in immune and inflammatory responses. Specifically binds to an IL-1 response element in the IL-6 gene. NF-IL6 also binds to regulatory regions of several acute-phase and cytokines genes. It probably plays a role in the regulation of acute-phase reaction, inflammation and hemopoiesis. The consensus recognition site is 5'-T[TG]NNGNAA[TG]-3'. Functions in brown adipose tissue (BAT) differentiation. -
組織特異性
Expressed at low levels in the lung, kidney and spleen. -
配列類似性
Belongs to the bZIP family. C/EBP subfamily.
Contains 1 bZIP domain. -
ドメイン
the 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors. -
翻訳後修飾
Sumoylated by polymeric chains of SUMO2 or SUMO3. -
細胞内局在
Nucleus. - Information by UniProt
-
参照データベース
- Entrez Gene: 1051 Human
- Entrez Gene: 12608 Mouse
- Entrez Gene: 24253 Rat
- Omim: 189965 Human
- SwissProt: P17676 Human
- SwissProt: P28033 Mouse
- SwissProt: P21272 Rat
- Unigene: 517106 Human
see all -
別名
- AGP/EBP antibody
- C EBP beta antibody
- C/EBP beta antibody
see all
画像
-
All lanes : Anti-CEBP Beta antibody [E299] - C-terminal (ab32358) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : CEBP Beta knockout HeLa cell lysate
Lane 3 : Jurkat cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 36 kDa
Observed band size: 40,45 kDa why is the actual band size different from the predicted?Lanes 1-3: Merged signal (red and green). Green - ab32358 observed at 40 and 45 kDa. Red - loading control ab7291 observed at 50 kDa.
ab32358 Anti-CEBP Beta antibody [E299] - C-terminal was shown to specifically react with CEBP Beta in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab261771 (knockout cell lysate ab256874) was used. Wild-type and CEBP Beta knockout samples were subjected to SDS-PAGE. ab32358 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 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 NIH/3T3 (Mouse embryonic fibroblast) cells labeling CEBP Beta with Purified ab32358 at 1:500 dilution (1.2 µg/ml). Cells were fixed in 100% Methanol. 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.
-
Anti-CEBP Beta antibody [E299] - C-terminal (ab32358) at 1/1000 dilution + NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates 15 at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 36 kDa -
Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling CEBP Beta with Purified ab32358 at 1/600 dilution (1µg/ml) (red). Cells were fixed with 80% 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 primary antibody and secondary antibody (Blue).
-
3T3-L1 (Mouse embryonic fibroblast cell line) cells were plated on cover slips, grown to post confluency and treated with adipogenic cocktail for 16 h. Cells were washed briefly with phosphate buffered saline (PBS) and fixed in methanol at −20°C for 5 min. Cells were then blocked with 1% BSA for 30 min before incubation with ab32358 at a 1/100 dilution at 4°C overnight and incubated with Alexa Fluor® 488 goat anti-rabbit secondary antibodies (1∶500) for 1 h at room temperature.
DAPI staining was used for visualizing the nuclei.
Images were acquired with an Olympus FlowView FV1000.
-
ab32358 (purified) at 1:30 dilution (2µg) immunoprecipitating CEBP Beta in NIH/3T3 whole cell lysate.
Lane 1 (input): NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10µg
Lane 2 (+): ab32358 & NIH/3T3 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab32358 in NIH/3T3 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. -
Anti-CEBP Beta antibody [E299] - C-terminal (ab32358) + MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 36 kDa -
Anti-CEBP Beta antibody [E299] - C-terminal (ab32358) at 1/1000 dilution + PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates 15ug at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 36 kDa -
Anti-CEBP Beta antibody [E299] - C-terminal (ab32358) at 1/1000 dilution + PC-12 (Rat adrenal gland pheochromocytoma cell line) cell lysate.
Predicted band size: 36 kDa
Observed bands
LAP*: 38kDa
LAP: 35kDa
LIP: 20kDa -
All lanes : Anti-CEBP Beta antibody [E299] - C-terminal (ab32358) at 1/500 dilution
Lane 1 : NIH/3T3 (Mouse embryo fibroblast cell line) cells
Lane 2 : MCF7 (Human breast adenocarcinoma cell line) cells
Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma cell line) cells
Lane 4 : Rat Spleen Lysate
Lane 5 : Rat Kidney Lysate
Lane 6 : Rat Heart Lysate
Lane 7 : Rat Brain Lysate
Lane 8 : Mouse Spleen Lysate
Lane 9 : Mouse Kidney Lysate
Lane 10 : Mouse Heart Lysate
Lane 11 : Mouse Brain Lysate
Predicted band size: 36 kDa -
ab32358, at a 1/50 dilution, staining CEBP Beta in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells by Immunofluorescence.
-
Overlay histogram showing HeLa (Human epithelial cell line from cervix adenocarcinoma) cells stained with ab32358 (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 (ab32358, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabeled 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.
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 (82)
ab32358 は 82 報の論文で使用されています。
- Zhang Q et al. Effects of 4-nonylphenol on adipogenesis in 3T3-L1 preadipocytes and C3H/10T1/2 mesenchymal stem cells. J Appl Toxicol 42:588-599 (2022). PubMed: 34553387
- Sterken BA et al. C/EBPβ isoform-specific regulation of migration and invasion in triple-negative breast cancer cells. NPJ Breast Cancer 8:11 (2022). PubMed: 35042889
- Zhang S et al. H3K27ac nucleosomes facilitate HMGN localization at regulatory sites to modulate chromatin binding of transcription factors. Commun Biol 5:159 (2022). PubMed: 35197580
- Song X et al. Jian Pi Tiao Gan Yin alleviates obesity phenotypes through mTORC1/SREBP1 signaling in vitro and in vivo. Ann Transl Med 10:291 (2022). PubMed: 35433951
- Müller C et al. Enhanced C/EBPβ function promotes hypertrophic versus hyperplastic fat tissue growth and prevents steatosis in response to high-fat diet feeding. Elife 11:N/A (2022). PubMed: 35451956