Name: Anti-CEBP Beta antibody [E299] - C-terminal
SKU: ab32358
Rating: 5 (7 reviews)

Anti-CEBP Beta antibody [E299] - C-terminal (ab32358) is a rabbit monoclonal antibody that is used to detect CEBP Beta in Western Blot, Flow Cytometry (Intra), Flow Cytometry, IP, ICC/IF. Suitable for Human, Mouse, Rat samples.

- Specificity confirmed with CEBP Beta knockout cell line validation

Images

Immunoprecipitation - Anti-CEBP Beta antibody [E299] - C-terminal (AB32358), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

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Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	ICC/IF	Flow Cyt (Intra)
Human	Expected	Tested	Tested	Tested
Mouse	Tested	Tested	Tested	Expected
Rat	Expected	Tested	Expected	Expected

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/30	Notes For unpurified use at 1/30

Expected
Expected

Species	Dilution info	Notes
Species Rat, Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000	Notes -
Species Rat	Dilution info 1/1000	Notes -
Species Human	Dilution info 1/1000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/500	Notes For unpurified use at 1/50 - 1/100
Species Human	Dilution info 1/500	Notes For unpurified use at 1/50 - 1/100

Expected
Expected

Species	Dilution info	Notes
Species Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/600	Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. For unpurified use at 1/1000

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Associated Products

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Target data

See full target information CEBPB

Function

Important transcription factor regulating the expression of genes involved in immune and inflammatory responses (PubMed:12048245, PubMed:1741402, PubMed:18647749, PubMed:9374525). Also plays a significant role in adipogenesis, as well as in the gluconeogenic pathway, liver regeneration, and hematopoiesis. The consensus recognition site is 5'-T[TG]NNGNAA[TG]-3'. Its functional capacity is governed by protein interactions and post-translational protein modifications. During early embryogenesis, plays essential and redundant roles with CEBPA. Has a promitotic effect on many cell types such as hepatocytes and adipocytes but has an antiproliferative effect on T-cells by repressing MYC expression, facilitating differentiation along the T-helper 2 lineage. Binds to regulatory regions of several acute-phase and cytokines genes and plays a role in the regulation of acute-phase reaction and inflammation. Also plays a role in intracellular bacteria killing (By similarity). During adipogenesis, is rapidly expressed and, after activation by phosphorylation, induces CEBPA and PPARG, which turn on the series of adipocyte genes that give rise to the adipocyte phenotype. The delayed transactivation of the CEBPA and PPARG genes by CEBPB appears necessary to allow mitotic clonal expansion and thereby progression of terminal differentiation (PubMed:20829347). Essential for female reproduction because of a critical role in ovarian follicle development (By similarity). Restricts osteoclastogenesis: together with NFE2L1; represses expression of DSPP during odontoblast differentiation (By similarity). Isoform 2. Essential for gene expression induction in activated macrophages. Plays a major role in immune responses such as CD4(+) T-cell response, granuloma formation and endotoxin shock. Not essential for intracellular bacteria killing. Isoform 3. Acts as a dominant negative through heterodimerization with isoform 2 (PubMed:11741938). Promotes osteoblast differentiation and osteoclastogenesis (By similarity).

Alternative names

TCF5, PP9092, CEBPB, CCAAT/enhancer-binding protein beta, C/EBP beta, Liver activator protein, Liver-enriched inhibitory protein, Nuclear factor NF-IL6, Transcription factor 5, LAP, LIP, TCF-5

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: E299
Purification technique: Affinity purification Protein A
Specificity: 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.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

What is this antibody validated in?

Anti-CEBP Beta antibody [E299] - C-terminal (ab32358) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Flow Cytometry (Intra), Flow Cytometry (Flow Cyt), Immunoprecipitation (IP), Immunocytochemistry/immunofluorescence (ICC/IF), in Human, Mouse, Rat samples.

What is the molecular weight of CEBP Beta?

Anti-CEBP Beta [E299] - C-terminal (ab32358) specifically detects a band for CEBP Beta (UniProt: P21272) at a molecular weight of 36kDa.

Recommended positive controls

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

Trusted by the scientific community

Anti-CEBP Beta [E299] - C-terminal (ab32358) was first used in a scientific publication in 2006 and has been cited over 100 times in peer-reviewed journals.

Reviewed by scientists

Anti-CEBP Beta [E299] - C-terminal (ab32358) has over 5 independent reviews from customers.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies. Specificity confirmed

The specificity of Anti-CEBP Beta antibody [E299] - C-terminal (ab32358) has been confirmed by Western blot testing in CEBPB Knockout HAP1 cells.

Other related products

We have a range of other formats of antibody clone [E299] also available for your convenience:
ab32358, Carrier free - ab220813, Alexa Fluor® 488 - ab237414, Alexa Fluor® 647 - ab237415

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

CEBP Beta also known as CCAAT/enhancer binding protein beta or C/EBPβ is a transcription factor that regulates gene expression in various tissues. It has a molecular mass of approximately 35 kDa. CEBP Beta is expressed in liver adipose tissue and immune cells among others. It binds to DNA at specific CCAAT motifs and regulates transcription of genes involved in immune response metabolism and differentiation.

Biological function summary

CEBP Beta plays important roles in cellular differentiation proliferation and immune function. CEBP Beta is part of the CEBP family of transcription factors which form homo- or hetero-dimers to function effectively often with other family members like CEBP Alpha. Its activity governs important processes in hematopoiesis and adipogenesis. This transcription factor is therefore essential in the regulation of energy homeostasis and immune response influencing the expression of various cytokines and acute-phase proteins.

Pathways

CEBP Beta integrates signals in key biological pathways such as the JAK-STAT and NF-kB pathways. Activation via cytokine signaling CEBP Beta interacts with STAT proteins to modulate gene expression linked to inflammatory responses. It has a significant role in the NF-kB pathway where it cooperates with Rel proteins to regulate genes involved in immune and stress responses including TNF and IL-1.

Associated diseases and disorders

CEBP Beta is implicated in conditions such as acute myeloid leukemia (AML) and obesity-related complications. Abnormal regulation of CEBP Beta can contribute to leukemogenesis by altering the expression of cell cycle and differentiation genes. Additionally CEBP Beta's role in lipid storage and insulin sensitivity links it to metabolic disorders such as Type 2 diabetes. Interactions with proteins like PPAR gamma in adipose tissues influence these disease processes.

Product promise

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12 product images

Immunoprecipitation - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358)
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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab32358 in NIH/3T3 whole cell lysate
For western blotting, VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used for detection at 1:1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST.
All lanes: Immunoprecipitation - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358)
Predicted band size: 36 kDa
Western blot - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358)
Lanes 1-3: Merged signal (red and green). Green - ab32358 observed at 40 and 45 kDa. Red - loading control Anti-alpha Tubulin antibody [DM1A] - 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 Human CEBPB (CEBP Beta) knockout HeLa cell line ab261771 (knockout cell lysate Human CEBPB (CEBP Beta) knockout HeLa 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 (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 (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (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.
All lanes: Western blot - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: CEBP Beta knockout HeLa cell lysate at 20 µg
Lane 2: Western blot - Human CEBPB (CEBP Beta) knockout HeLa cell line (Human CEBPB (CEBP Beta) knockout HeLa cell line ab261771)
Lane 3: Jurkat cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (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 kDa, 45 kDa
Western blot - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358)
All lanes: Western blot - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358) at 1/1000 dilution
All lanes: NIH/3T3 (Mouse embryonic fibroblast) whole cell lysates 15 at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 36 kDa
Flow Cytometry (Intracellular) - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358)
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, Goat Anti-Rabbit IgG H&L (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).
Immunocytochemistry/ Immunofluorescence - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358)
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 Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1:200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, Goat Anti-Rabbit IgG H&L (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.
Image from Sikkeland et al. PLoS One. 2013 Jul 10;8(7):e68249. doi: 10.1371/journal.pone.0068249. Print 2013. Fig S1.
Immunocytochemistry/ Immunofluorescence - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358)
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.
Western blot - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358)
All lanes: Western blot - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358)
All lanes: MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 36 kDa
Western blot - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358)
All lanes: Western blot - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358) at 1/1000 dilution
All lanes: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates 15ug at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 36 kDa
Flow Cytometry (Intracellular) - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358)
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) (Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed 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.
Western blot - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358)
Observed bands

LAP*: 38kDa

LAP: 35kDa

LIP: 20kDa
All lanes: Western blot - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358) at 1/1000 dilution
All lanes: PC-12 (Rat adrenal gland pheochromocytoma cell line) cell lysate.
Predicted band size: 36 kDa
Western blot - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358)
All lanes: Western blot - 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
Immunocytochemistry/ Immunofluorescence - Anti-CEBP Beta antibody [E299] - C-terminal (ab32358)
ab32358, at a 1/50 dilution, staining CEBP Beta in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells by Immunofluorescence.