Validated using a knockout cell line
Recombinant
RabMAb

Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Loading Control (HRP) (ab194109)

Overview

  • Product name
    Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Loading Control (HRP)
    See all Lamin B1 primary antibodies
  • Description
    Rabbit monoclonal [EPR8985(B)] to Lamin B1 - Nuclear Loading Control (HRP)
  • Host species
    Rabbit
  • Conjugation
    HRP
  • Tested applications
    Suitable for: WB, IHC-Pmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Mouse, Rat
  • Immunogen

    Synthetic peptide. within Human Lamin B1 aa 500 to the C-terminus (internal sequence). The exact sequence is proprietary.
    Database link: P20700

  • Positive control
    • WB: Jurkat, MOLT4, Y79, CaCo 2 whole cell lysates. IHC: normal human colon tissue.
  • General notes

    Alternative versions available:
    Anti-Lamin B1 antibody [EPR8985(B)] (ab133741) - Knockout validated

    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 ab194109 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 1/5000. Detects a band of approximately 70 kDa (predicted molecular weight: 66 kDa).
IHC-P 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

ab199507 - Rabbit monoclonal IgG (HRP), is suitable for use an as isotype control with this antibody.

Target

  • Function
    Lamins are components of the nuclear lamina, a fibrous layer on the nucleoplasmic side of the inner nuclear membrane, which is thought to provide a framework for the nuclear envelope and may also interact with chromatin.
  • Involvement in disease
    Defects in LMNB1 are the cause of leukodystrophy demyelinating autosomal dominant adult-onset (ADLD) [MIM:169500]. ADLD is a slowly progressive and fatal demyelinating leukodystrophy, presenting in the fourth or fifth decade of life. Clinically characterized by early autonomic abnormalities, pyramidal and cerebellar dysfunction, and symmetric demyelination of the CNS. It differs from multiple sclerosis and other demyelinating disorders in that neuropathology shows preservation of oligodendroglia in the presence of subtotal demyelination and lack of astrogliosis.
  • Sequence similarities
    Belongs to the intermediate filament family.
  • Post-translational
    modifications
    B-type lamins undergo a series of modifications, such as farnesylation and phosphorylation. Increased phosphorylation of the lamins occurs before envelope disintegration and probably plays a role in regulating lamin associations.
  • Cellular localization
    Nucleus inner membrane.
  • Information by UniProt
  • Database links
  • Alternative names
    • ADLD antibody
    • lamin B1 antibody
    • Lamin-B1 antibody
    • LMN antibody
    • LMN2 antibody
    • LMNB antibody
    • Lmnb1 antibody
    • LMNB1_HUMAN antibody
    • MGC111419 antibody
    • OTTHUMP00000159218 antibody
    see all

Images

  • All lanes : Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Loading Control (HRP) (ab194109) at 1/5000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : LMNB1 (Lamin B1) knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 66 kDa
    Observed band size: 70 kDa (why is the actual band size different from the predicted?)


    Exposure time: 3 minutes


    ab194109 was shown to specifically react with Lamin B1 in wild-type HAP1 cells as signal was lost in LMNB1 (Lamin B1) knockout cells. Wild-type and LMNB1 (Lamin B1) knockout samples were subjected to SDS-PAGE. Ab194109 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/20000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

  • IHC image of Lamin B1 staining in a section of formalin-fixed paraffin-embedded normal human colon tissue*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab194109 at 1/500 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • All lanes : Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Loading Control (HRP) (ab194109) at 1/5000 dilution

    Lane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 2 : MOLT4 (Human acute lymphoblastic leukemia cell line) Whole Cell Lysate
    Lane 3 : Y79 (Human retinoblastoma cell line) Whole Cell Lysate
    Lane 4 : Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 66 kDa
    Observed band size: 70 kDa (why is the actual band size different from the predicted?)


    Exposure time: 1 minute


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab194109 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

References

This product has been referenced in:
  • Zeng L  et al. A Novel EphA2 Inhibitor Exerts Beneficial Effects in PI-IBS in Vivo and in Vitro Models via Nrf2 and NF-?B Signaling Pathways. Front Pharmacol 9:272 (2018). Read more (PubMed: 29662452) »
  • Xu Q  et al. Paeonol Reduces the Nucleocytoplasmic Transportation of HMGB1 by Upregulating HDAC3 in LPS-Induced RAW264.7 Cells. Inflammation N/A:N/A (2018). WB . Read more (PubMed: 29736733) »

See all 4 Publications for this product

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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