Recombinant
RabMAb

Recombinant Anti-CEBP Beta antibody [E299] - C-terminal (ab32358)

Overview

  • Product name
    Anti-CEBP Beta antibody [E299] - C-terminal
    See all CEBP Beta primary antibodies
  • Description
    Rabbit monoclonal [E299] to CEBP Beta - C-terminal
  • Host species
    Rabbit
  • 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.
  • Tested applications
    Suitable for: EMSA, WB, ICC/IF, Flow Cyt, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Rat CEBP Beta aa 250-350 (C terminal). The exact sequence is proprietary.
    Database link: P21272

  • Positive control
    • WB: PC-12, NIH/3T3 and MCF7 cell lysate. ICC/IF: HeLa and NIH/3T3 cells. Flow Cyt: HeLa cells. IP: NIH/3T3 cell lysate
  • General notes

    A trial size is available to purchase for this antibody.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab32358 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
EMSA Use at an assay dependent concentration. PubMed: 18462018
WB 1/1000. Predicted molecular weight: 36 kDa.
ICC/IF 1/500.

For unpurified use at 1/50 - 1/100

 

Flow Cyt 1/600.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

For unpurified use at 1/1000

IP 1/30.

For unpurified use at 1/30

Target

  • Function
    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.
  • Tissue specificity
    Expressed at low levels in the lung, kidney and spleen.
  • Sequence similarities
    Belongs to the bZIP family. C/EBP subfamily.
    Contains 1 bZIP domain.
  • Domain
    the 9aaTAD motif is a transactivation domain present in a large number of yeast and animal transcription factors.
  • Post-translational
    modifications
    Sumoylated by polymeric chains of SUMO2 or SUMO3.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • AGP/EBP antibody
    • C EBP beta antibody
    • C/EBP beta antibody
    • C/EBP related protein 2 antibody
    • CCAAT Enhancer Binding Protein beta antibody
    • CCAAT/enhancer-binding protein beta antibody
    • CEBPB antibody
    • CEBPB_HUMAN antibody
    • CRP2 antibody
    • IL 6DBP antibody
    • IL6DBP antibody
    • Interleukin 6 dependent binding protein antibody
    • LAP antibody
    • Liver activator protein antibody
    • Liver enriched transcriptional activator antibody
    • NF IL6 antibody
    • NFIL6 antibody
    • Nuclear factor NF IL6 antibody
    • Nuclear factor NF-IL6 antibody
    • SF B antibody
    • SFB antibody
    • Silencer factor B antibody
    • TCF-5 antibody
    • TCF5 antibody
    • Transcription factor 5 antibody
    see all

Images

  • 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

  • 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.

  • 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.

  • 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).

  • 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.

References

This product has been referenced in:
  • Welz B  et al. Proteome and Phosphoproteome Analysis in TNF Long Term-Exposed Primary Human Monocytes. Int J Mol Sci 20:N/A (2019). Read more (PubMed: 30871024) »
  • Guo Q  et al. C/EBPß mediates palmitate-induced musclin expression via the regulation of PERK/ATF4 pathways in myotubes. Am J Physiol Endocrinol Metab 316:E1081-E1092 (2019). Read more (PubMed: 30964708) »
See all 25 Publications for this product

Customer reviews and Q&As

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1-5 of 5 Abreviews

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Non-reduced Denaturing (10% gel)
Sample
Mouse Cell lysate - whole cell (C3H10T cells)
Specification
C3H10T cells
Treatment
25ng/ml (BMP4)
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 22°C

Abcam user community

Verified customer

Submitted Mar 26 2015

Application
Western blot
Sample
Human Cell lysate - whole cell (mcf7)
Loading amount
100000 cells
Specification
mcf7
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted May 10 2012

Application
Western blot
Sample
Human Cell lysate - other (HT1080 Cell line)
Loading amount
100000 cells
Specification
HT1080 Cell line
Gel Running Conditions
Reduced Denaturing (12% SDS-PAGE)
Blocking step
Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Dr. Neil Taylor

Verified customer

Submitted Feb 08 2010

Application
Immunocytochemistry
Sample
Mouse Cultured Cells (Mouse Embryonic Fibroblasts)
Specification
Mouse Embryonic Fibroblasts
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100

Dr. Neil Taylor

Verified customer

Submitted Oct 06 2009

Application
Immunocytochemistry
Sample
Human Cultured Cells (HT1080 Cell line)
Specification
HT1080 Cell line
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.1% Triton X-100

Dr. Neil Taylor

Verified customer

Submitted Oct 06 2009

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