Hello. We're improving abcam.com and we'd welcome your feedback.

Hello. We're improving abcam.com and we'd welcome your feedback.

Infomation icon

We haven't added this to the BETA yet

New BETA website

New BETA website

Hello. We're improving abcam.com and we'd welcome your feedback.

Take a look at our BETA site and see what we’ve done so far.

Switch on our new BETA site

Now available

Search and browse selected products

  • A selection of primary antibodies

Purchase these through your usual distributor

In the coming months

  • Additional product types
  • Supporting content
  • Sign in to your account
  • Purchase online
Validated using a knockout cell lineRecombinantRabMAb

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

Submit a review Submit a question
Western blot - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)
  • Immunocytochemistry/ Immunofluorescence - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)
  • Immunocytochemistry/ Immunofluorescence - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)
  • Western blot - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)
  • Flow Cytometry (Intracellular) - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)
  • Immunoprecipitation - Anti-Lin28B antibody [EPR18717] - BSA and Azide free (ab240326)
  • 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: Flow Cyt (Intra), WB, IP, ICC/IF
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Unconjugated

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: Flow Cyt (Intra), WB, 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 (intra): K562 cells. IP: K562 cell lysate.

  • General notes

    ab240326 is the carrier-free version of ab191881.

    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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab240326 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
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 30 kDa (predicted molecular weight: 27 kDa).
IP
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 30 kDa (predicted molecular weight: 27 kDa).
IP
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.

Target

  • Function

    Acts as a suppressor of microRNA (miRNA) biogenesis by specifically binding the precursor let-7 (pre-let-7), a miRNA precursor. Acts by binding pre-let-7 and recruiting ZCCHC11/TUT4 uridylyltransferase, leading to the terminal uridylation of pre-let-7. Uridylated pre-let-7 miRNAs fail to be processed by Dicer and undergo degradation. Specifically recognizes the 5'-GGAG-3' motif in the terminal loop of pre-let-7. Also recognizes and binds non pre-let-7 pre-miRNAs that contain the 5'-GGAG-3' motif in the terminal loop, leading to their terminal uridylation and subsequent degradation. Mediates MYC-mediated let-7 repression. Isoform 1, when overexpressed, stimulates growth of the breast adenocarcinoma cell line MCF-7. Isoform 2 has no effect on cell growth.

  • Tissue specificity

    High expression in testis, fetal liver, placenta and in hepatocellular carcinoma (HCC). Isoform 1 is only detected in moderately and poorly differentiated HCC tissues and placenta (at protein level). Isoform 2 is detected in fetal liver, non-tumor liver tissues, as well as well-differentiated tumor tissues (at protein level).

  • Sequence similarities

    Belongs to the lin-28 family.
    Contains 2 CCHC-type zinc fingers.
    Contains 1 CSD (cold-shock) domain.

  • Cellular localization

    Cytoplasm. Nucleus. Predominantly cytoplasmic at G1 phase, accumulates in the nucleus in S and G2 phases. The frequency of nuclear localization in S and in G2 phases is 60% and 30%, respectively.
  • Information by UniProt

  • Database links

    • Alternative names

      • CSDD 2 antibody
      • CSDD2 antibody
      • FLJ16517 antibody
      • Lin 28 homolog B (C. elegans) antibody
      • Lin 28.2 antibody
      • Lin-28B antibody
      • LIN28B antibody
      • LN28B_HUMAN antibody
      • Protein lin-28 homolog B antibody
      see all

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

      Intracellular 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)

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    References (0)

    Publishing research using ab240326? Please let us know so that we can cite the reference in this datasheet.

    ab240326 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    There are currently no Customer reviews or Questions for ab240326.
    Please use the links above to contact us or submit feedback about this product.

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