Astana Biomed Group, an authorized Abcam distributor in Central Asia

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, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

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, Flow Cyt, ICC/IFmore details
    Unsuitable for: IHC-P

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    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: HeLa cells. IP: NIH/3T3 cell lysate.

  • 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

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

    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.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

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

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

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Alternative products to Anti-CEBP Beta antibody [E299] - BSA and Azide free (ab220813)

© 2021 Astana Biomed Group LLP. All rights reserved.