重组Anti-CEBP Beta抗体[E299] - BSA and Azide free (ab220813)
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 - BSA and Azide free
- Suitable for: WB, IP, ICC/IF, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-CEBP Beta抗体[E299] - BSA and Azide free
参阅全部 CEBP Beta 一抗 -
描述
兔单克隆抗体[E299] to CEBP Beta - BSA and Azide free -
宿主
Rabbit -
特异性
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.
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经测试应用
适用于: WB, IP, ICC/IF, Flow Cyt (Intra)more details
不适用于: IHC-P -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab274870) -
阳性对照
- WB: HeLa, Jurkat and PC12 cell lysates. IF: HeLa, NIH/3T3 and 3T3-L1 cells. Flow Cyt (intra): HeLa cells. IP: NIH/3T3 cell lysate.
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常规说明
ab220813 is the carrier-free version of ab32358.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
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.
性能
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形式
Liquid -
存放说明
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
存储溶液
pH: 7.20
Constituent: PBS -
无载体
是 -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
E299 -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab220813于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。
应用 | Ab评论 | 说明 |
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WB |
Use at an assay dependent concentration. Predicted molecular weight: 36 kDa.
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IP |
Use at an assay dependent concentration.
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ICC/IF |
Use at an assay dependent concentration.
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Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
说明 |
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WB
Use at an assay dependent concentration. Predicted molecular weight: 36 kDa. |
IP
Use at an assay dependent concentration. |
ICC/IF
Use at an assay dependent concentration. |
Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
靶标
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功能
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. -
组织特异性
Expressed at low levels in the lung, kidney and spleen. -
序列相似性
Belongs to the bZIP family. C/EBP subfamily.
Contains 1 bZIP domain. -
结构域
the 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors. -
翻译后修饰
Sumoylated by polymeric chains of SUMO2 or SUMO3. -
细胞定位
Nucleus. - Information by UniProt
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数据库链接
- 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 -
别名
- AGP/EBP antibody
- C EBP beta antibody
- C/EBP beta antibody
see all
图片
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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?This data was developed using the same antibody clone in a different buffer formulation (ab32358).
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.
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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.This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32358).
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Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling CEBP Beta with Purified ab32358 at 1/600 dilution (1 µg/ml) (red). Cells were fixed with 80% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 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).
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32358).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32358).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32358).
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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.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32358).
实验方案
数据表及文件
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Datasheet download
Certificate of Compliance
文献 (9)
ab220813 被引用在 9 文献中.
- Guaraldo M et al. Characterization of human mitochondrial ferritin promoter: identification of transcription factors and evidences of epigenetic control. Sci Rep 6:33432 (2016). ChIP ; Human . PubMed: 27625068
- Viart V et al. Transcription factors and miRNAs that regulate fetal to adult CFTR expression change are new targets for cystic fibrosis. Eur Respir J 45:116-28 (2015). PubMed: 25186262
- Xu S et al. LIFRa-CT3 induces differentiation of a human acute myelogenous leukemia cell line HL-60 by suppressing miR-155 expression through the JAK/STAT Pathway. Leuk Res N/A:N/A (2014). Human . PubMed: 25092123
- Myles IA et al. Signaling via the IL-20 receptor inhibits cutaneous production of IL-1ß and IL-17A to promote infection with methicillin-resistant Staphylococcus aureus. Nat Immunol 14:804-11 (2013). WB, ChIP . PubMed: 23793061
- Sikkeland J & Saatcioglu F Differential expression and function of stamp family proteins in adipocyte differentiation. PLoS One 8:e68249 (2013). ICC/IF . PubMed: 23874564
- Ratajewski M et al. ABCC6 Expression Is Regulated by CCAAT/Enhancer-Binding Protein Activating a Primate-Specific Sequence Located in the First Intron of the Gene. J Invest Dermatol : (2012). PubMed: 22763786
- Michas G et al. Reciprocal regulation of 11ß-hydroxysteroid dehydrogenase 1 and glucocorticoid receptor expression by dexamethasone inhibits human coronary artery smooth muscle cell proliferation in vitro. Mol Cell Biochem 346:69-79 (2011). WB ; Human . PubMed: 20922465
- Pal R et al. C/EBPbeta regulates transcription factors critical for proliferation and survival of multiple myeloma cells. Blood 114:3890-8 (2009). WB, ChIP ; Human . PubMed: 19717648
- Meyer KB et al. Allele-Specific Up-Regulation of FGFR2 Increases Susceptibility to Breast Cancer. PLoS Biol 6:e108 (2008). EMSA ; Human . PubMed: 18462018