Recombinant
RabMAb

Recombinant Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)

Rabbit recombinant monoclonal CEBP Beta antibody [E299]. Validated in WB, IP, EMSA, Flow Cyt, ICC/IF and tested in Mouse, Rat, Human. Cited in 9 publication(s).

Overview

  • Product name

    Anti-CEBP Beta antibody [E299] - BSA and Azide free
    See all CEBP Beta primary antibodies
  • Description

    Rabbit monoclonal [E299] to CEBP Beta - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IP, EMSA, Flow Cyt, ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to residues in the C terminus of Rat CEBPB.

  • Positive control

    • WB: PC12 cell lysate. IF: Hela cells.
  • General notes

    Ab220813 is the carrier-free version of ab32358. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    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.

    ab220813 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab220813 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
WB Use at an assay dependent concentration. Predicted molecular weight: 36 kDa.
IP Use at an assay dependent concentration.
EMSA Use at an assay dependent concentration. PubMed: 18462018
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for IHC-P.
  • 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

    • 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).

    • 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).

    • 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).

    • 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).

    • 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).

    • ab32358, at a 1/50 dilution, staining CEBP Beta in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells by Immunofluorescence.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32358).

    References

    This product has been referenced in:

    • 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 . Read more (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). Read more (PubMed: 25186262) »
    See all 9 Publications for this product

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