Overview

  • Product name

    Anti-Lamin B1 antibody [119D5-F1]
    See all Lamin B1 primary antibodies
  • Description

    Mouse monoclonal [119D5-F1] to Lamin B1
  • Host species

    Mouse
  • Specificity

    Reacts with an epitope located C-terminal of residue 231 in lamin B1. Reacts against lamin B1, does not cross react with lamin B2. Lamins do not appear to be universally distributed among different cell and tissue types. ab8982 has been shown to react with HeLa and 3T3 cells in immunocytochemistry. Other cell/tissue types have not been tested.
  • Tested applications

    Suitable for: WB, Dot blot, ELISA, Flow Cyt, ICC, ICC/IF, IHC-Frmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Sheep, Rabbit, Cow, Dog, Human
  • Immunogen

    Tissue, cells or virus corresponding to Rat Lamin B1 (C terminal). Immunogen = Purified Rat liver lamins

  • Epitope

    C-terminal to residue 231.
  • Positive control

    • NIH 3T3 whole cell lysate (ab7179) can be used as a positive control in WB. ICC/IF: Wildtype HAP1 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab8982 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 1/100 - 1/1000. Predicted molecular weight: 67 kDa.
Dot blot Use at an assay dependent concentration.
ELISA Use at an assay dependent concentration.
Flow Cyt 1/100 - 1/200.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ICC 1/20 - 1/50.
ICC/IF Use a concentration of 0.5 µg/ml.
IHC-Fr Use at an assay dependent concentration.

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 [119D5-F1] (ab8982) at 1/2000 dilution

    Lane 1 : Whole cell lysate prepared from human mammary cells (24 hour culture)
    Lane 2 : Whole cell lysate prepared from human mammary cells (48 hour culture)

    Lysates/proteins at 50000 cells per lane.

    Secondary
    All lanes : HRP conjugated goat anti-mouse polyclonal at 1/5000 dilution

    Developed using the ECL technique.

    Predicted band size: 67 kDa
    Observed band size: 70 kDa
    why is the actual band size different from the predicted?


    Exposure time: 40 seconds

    See Abreview

  • Methanol fixed HeLa and 3T3 cells (ab7179) were stained with ab8982 (1/50 and 1/20 respectively). The cells were fixed in 100% methanol for 6 minutes at -20°C. ab8982 clearly stains the nuclear envelope with very little backgrouns staining. 

    A: HeLa cells + ab8982 (1/50) (green)
    B: HeLa cells counterstained with DAPI (blue)
    C. 3T3 cells + ab8982 (1/20) (green)
    D. 3T3 cells counterstained with DAPI (blue)

  • ab8982 staining Lamin B1 in wild-type HAP1 cells (top panel) and LMNB1 knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab8982 at 0.5μg/ml and ab202272 at 1/250 dilution (shown in pseudocolour red) overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to Mouse IgG (Alexa Fluor® 488) (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.

    Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

  • Overlay histogram showing HeLA cells stained with ab8982 (red line). The cells were fixed with 100% 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 (ab7481) / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab8982, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was Mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

  • Immunohistochemistry on frozen sections of swine liver showing nuclear lamina staining in hepatocytes
  • Immunohistochemistry on frozen sections of human colon showing nuclear lamina staining in epithelial and connective tissue cells

References

This product has been referenced in:

  • Lukášová E  et al. Distinct cellular responses to replication stress leading to apoptosis or senescence. FEBS Open Bio 9:870-890 (2019). Read more (PubMed: 30982228) »
  • Paonessa F  et al. Microtubules Deform the Nuclear Membrane and Disrupt Nucleocytoplasmic Transport in Tau-Mediated Frontotemporal Dementia. Cell Rep 26:582-593.e5 (2019). Read more (PubMed: 30650353) »
See all 25 Publications for this product

Customer reviews and Q&As

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1-4 of 4 Abreviews

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Tri-Sodium Citrate
Permeabilization
Yes - with blocking
Specification
Brain
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Juliana Schneider

Verified customer

Submitted Mar 28 2017

Application
Western blot
Loading amount
15 µg
Gel Running Conditions
Non-reduced Denaturing (8%)
Sample
Human Tissue lysate - whole (Brain)
Specification
Brain
Blocking step
5% BSA + 5% Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C

Abcam user community

Verified customer

Submitted Oct 27 2014

Application
Western blot
Sample
Human Cell lysate - whole cell (Human mammary cell)
Loading amount
50000 cells
Specification
Human mammary cell
Gel Running Conditions
Reduced Denaturing (8%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Feb 22 2012

Application
Western blot
Sample
Human Cell lysate - whole cell (Colon cell line)
Loading amount
20 µg
Specification
Colon cell line
Gel Running Conditions
Reduced Denaturing (4-12%)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Dec 30 2010

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