Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR18717] to Lin28B - BSA and Azide free
  • Suitable for: WB, Flow Cyt, IP, ICC/IF
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Lin28B antibody [EPR18717] - BSA and Azide free
    See all Lin28B primary antibodies

  • Description

    Rabbit monoclonal [EPR18717] to Lin28B - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WB, Flow Cyt, IP, ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • WB: HEK293, K562 and HepG2 cell lysates. ICC: JAR and K562 cells. Flow Cyt: K562 cells. IP: K562 cell lysate.

  • General notes

    Ab240326 is the carrier-free version of ab191881. 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

    ab240326 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Images

  • Western blot - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)
    Western blot - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)
    All lanes : Anti-Lin28B antibody [EPR18717] (ab191881) at 1/2000 dilution

    Lane 1 : Wild-type HEK293 cell lysate
    Lane 2 : LIN28B knockout HEK293 cell lysate
    Lane 3 : HepG2 cell lysate
    Lane 4 : K562 cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 27 kDa
    Observed band size: 35 kDa
    why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab191881).

    Lanes 1 - 4: Merged signal (red and green). Green - ab191881 observed at 35 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55 kDa.

    ab191881 was shown to react with Lin28B in wild-type HEK-293 cells in Western blot with loss of signal observed in LIN28B knockout sample. Wild-type HEK-293 and LIN28B knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween®) before incubation with ab191881 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 2000 dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)
    Immunocytochemistry/ Immunofluorescence - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized JAR (Human placenta choriocarcinoma cell line) cells labeling Lin28B with ab191881 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasm and nuclear staining on JAR cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab191881 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191881).

  • Immunocytochemistry/ Immunofluorescence - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)
    Immunocytochemistry/ Immunofluorescence - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)

    Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling Lin28B with ab191881 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing cytoplasm and nuclear staining on K562 cells. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution (red).

    The negative controls are as follows:
    -ve control 1: ab191881 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/1000 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/1000 dilution.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191881).

  • Western blot - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)
    Western blot - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)
    All lanes : Anti-Lin28B antibody [EPR18717] (ab191881) at 1/1000 dilution

    Lane 1 : Wild-type HEK293T cell lysate
    Lane 2 : LIN28B knockout HEK293T cell lysate
    Lane 3 : HepG2 cell lysate
    Lane 4 : SW480 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 27 kDa
    Observed band size: 36 kDa why is the actual band size different from the predicted?



    This data was developed using the same antibody clone in a different buffer formulation (ab191881).

    Lanes 1-4: Merged signal (red and green). Green - ab191881 observed at 36 kDa. Red - loading control ab7291 observed at 50 kDa.

    ab191881 Anti-Lin28B antibody [EPR18717] was shown to specifically react with Lin28B in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab265066 (knockout cell lysate ab257504) was used. Wild-type and Lin28B knockout samples were subjected to SDS-PAGE. ab191881 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.

     

     

  • Flow Cytometry - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)
    Flow Cytometry - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)

    Flow cytometric analysis of 4% paraformaldehyde-fixed K562 (Human chronic myelogenous leukemia cells from bone marrow) cells labeling Lin28B with ab191881 at 1/150 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191881).

  • Immunoprecipitation - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)
    Immunoprecipitation - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)

    Lin28B was immunoprecipitated from 1mg of K562 (Human chronic myelogenous leukemia cells from bone marrow) whole cell lysate with ab191881 at 1/50 dilution. Western blot was performed from the immunoprecipitate using ab191881 at 1/1000 dilution. Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500 dilution.

    Lane 1: K562 whole cell lysate 10ug (Input).

    Lane 2: ab191881 IP in K562 whole cell lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab191881 in K562 whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

    Exposure time: 5 seconds.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab191881).

  • Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)
    Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)

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