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  1. Link

    cebp-beta-antibody-e299-bsa-and-azide-free-ab220813.pdf

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Epigenetics and Nuclear Signaling Transcription Other factors
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Validated using a knockout cell lineRecombinantRabMAb

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

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  • Certificate of Compliance
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Western blot - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)
  • Immunoprecipitation - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)
  • Flow Cytometry (Intracellular) - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)
  • Immunocytochemistry/ Immunofluorescence - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)
  • Immunocytochemistry/ Immunofluorescence - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)
  • Flow Cytometry (Intracellular) - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)
  • Immunocytochemistry/ Immunofluorescence - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)
  • Anti-CEBP Beta antibody [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

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 Unconjugated

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Knockout
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Human CEBPB (CEBP Beta) knockout HeLa cell line (ab261771)

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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, ICC/IF, Flow Cyt (Intra)more details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
    (Peptide available as ab274870)

  • Positive control

    • 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.
  • General notes

    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
    For more information see here.

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

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C. Do Not Freeze.
  • Storage buffer

    pH: 7.20
    Constituent: PBS
  • Carrier free

    Yes
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    E299
  • Isotype

    IgG
  • Research areas

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Other factors
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Transcription Factors
    • Cardiovascular
    • Atherosclerosis
    • Diabetes associated
    • Cardiovascular
    • Heart
    • Hypertrophy
    • Transcription factors

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-CEBP Beta antibody [E299] - C-terminal (ab237414)
    • Alexa Fluor® 647 Anti-CEBP Beta antibody [E299] (ab237415)
    • Anti-CEBP Beta antibody [E299] - C-terminal (ab32358)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat Anti-Rabbit IgG H&L (DyLight® 488) preadsorbed (ab96899)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Immunizing Peptide (Blocking)

    • Anti-CEBP Beta antibody [E299] - C-terminal Blocking Peptide (ab274870)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human CEBPB (CEBP Beta) knockout HeLa cell line (ab261771)
  • KO cell lysates

    • Human CEBPB (CEBP Beta) knockout HeLa cell lysate (ab256874)
  • Positive Controls

    • PC-12 nuclear extract lysate (ab14884)

Applications

The Abpromise guarantee

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.Can be blocked with CEBP Beta peptide (ab274870).
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.

Notes
WB
Use at an assay dependent concentration. Predicted molecular weight: 36 kDa.Can be blocked with CEBP Beta peptide (ab274870).
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.

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.
  • Target information above from: UniProt accession P17676 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • 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
    • Unigene: 719041 Human
    • Unigene: 720603 Human
    • Unigene: 439656 Mouse
    • Unigene: 6479 Rat
    see all
  • 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

  • Western blot - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)
    Western blot - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)
    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.

    Performed under reducing conditions.

    Predicted band size: 36 kDa



    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.

     

     

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

    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 (Intracellular) - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)
    Flow Cytometry (Intracellular) - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)

    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). 

     

     

  • Immunocytochemistry/ Immunofluorescence - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)
    Immunocytochemistry/ Immunofluorescence - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)

    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)
    Immunocytochemistry/ Immunofluorescence - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)Image from Sikkeland et al. PLoS One. 2013 Jul 10;8(7):e68249. doi: 10.1371/journal.pone.0068249. Print 2013. Fig S1.

    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).

  • Flow Cytometry (Intracellular) - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)
    Flow Cytometry (Intracellular) - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)

    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). 

     

     

  • Immunocytochemistry/ Immunofluorescence - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)
    Immunocytochemistry/ Immunofluorescence - Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)

    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).

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

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (9)

Publishing research using ab220813? Please let us know so that we can cite the reference in this datasheet.

ab220813 has been referenced in 9 publications.

  • 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

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