Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Lamin B1 antibody [EPR8985(B)] (ab133741)

Overview

  • Product name

    Anti-Lamin B1 antibody [EPR8985(B)]
    See all Lamin B1 primary antibodies
  • Description

    Rabbit monoclonal [EPR8985(B)] to Lamin B1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IP, ICC/IF, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Positive control

    • WB: Jurkat, Molt4, Y79, Caco 2, C6, Raw264.7, PC-12 and NIH/3T3 cell lysates. Mouse brain,heart, kidney and spleen; and Rat brain, heart and spleen lysates. IHC-P: Human colon, liver and transitional cell carcinoma of the bladder tissues. ICC/IF: Ramos cells, HAP1-LMNB1 cells.
  • General notes

    A trial size is available to purchase for this antibody.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab133741 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
IP 1/20.
ICC/IF Use a concentration of 1 µg/ml.
WB 1/1000 - 1/10000. Detects a band of approximately 70 kDa (predicted molecular weight: 66 kDa).
IHC-P 1/300. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

See IHC antigen retrieval protocols.

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

  • Lane 1: Hap1 wildtype cell lysate (20 µg)

    Lane 2: LMNB1 Hap1 knockout cell lysate (20 µg)

    Lane 3: HeLa wildtype cell lysate (20 µg)

    Lane 4: LMNB1 HeLa knockout cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab133741 observed at 66-70 kDa. Red - loading control, ab8245 observed at 37 kDa.

    ab133741 was shown to react with Lamin B1 in HeLa wildtype. Loss of signal was observed when knockout sample ab263825 was used. Wild-type and Lamin B1 knockout samples were subjected to SDS-PAGE. ab133741 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) 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.
  • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: Empty
    Lane 3: LMNB1 HAP1 knockout whole cell lysate (20 µg)
    Lane 4: Empty
    Lanes 1 - 4: Merged signal (red and green). Green - ab133741 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab133741 was shown to specifically react with Lamin B1 in wild type HAP1 cells. No band was observed when knockout samples were used. Wild-type and Lamin B1 knockout samples were subjected to SDS-PAGE. Ab133741 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed 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 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Ramos (Human Burkitt's lymphoma cell line) cells labeling Lamin B1 with ab133741 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/200 dilution (green). Nuclear envelope staining on Ramos cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
    The negative controls are as follows:
    -ve control 1: ab133741 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
    -ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/200 dilution.

     

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human transitional cell carcinoma of the bladder tissue labeling Lamin B1 with purified ab133741 at 1/300. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

  • ab133741 staining Lamin B1 in wild-type HAP1 cells (top panel) and Lamin B1 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 ab133741 at 1μg/ml dilution and ab195889 at 1/250 dilution (shown in pseudo-color red) overnight at +4°C. The cells were then incubated with ab150081 (Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488)) at 1/1000 dilution for 1 hour. Nuclear DNA was labelled in blue with DAPI.

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

  • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: Empty
    Lane 3: Lamin B1 knockout HAP1 cell lysate (20 µg)
    Lanes 1 - 3: Merged signal (red and green).

    Green - Target observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.

    This western blot image is a comparison between ab133741 and a competitor's discontinued goat polyclonal antibody.

     

  • Anti-Lamin B1 antibody [EPR8985(B)] (ab133741) at 1/1000 dilution + GST-tagged Recombinant Human Lamin B1 protein (aa 1 to 586) at 0.015 µg with 5% NFDM/TBST

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

    Predicted band size: 66 kDa
    Observed band size: 100 kDa
    why is the actual band size different from the predicted?


    Exposure time: 1 second


    Recombinant Human Lamin B1 protein (ab114163)

  • All lanes : Anti-Lamin B1 antibody [EPR8985(B)] (ab133741) at 1/50000 dilution (purified)

    Lane 1 : Jurkat (Human T cell leukemia cells from peripheral blood)
    cell lysate
    Lane 2 : Molt-4 (Human lymphoblastic leukemia cell line) cell lysate
    Lane 3 : Y79 (Human retinoblastoma cell line) cell lysate
    Lane 4 : Caco-2 (Human colorectal adenocarcinoma cells) cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution

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



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunohistochemical staining of paraffin embedded Mouse Cerebral cortex with purified ab133741 at a working dilution of 1/300. The secondary antibody used is a HRP polymer for rabbit IgG. Nuclear envelope staining on neuron cells of Cerebral cortex tissue is observed. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

  • All lanes : Anti-Lamin B1 antibody [EPR8985(B)] (ab133741) at 1/50000 dilution (purified)

    Lane 1 : C6 (Rat glial tumor cells) cell lysate
    Lane 2 : PC12 (Rat adrenal gland pheochromocytoma) cell lysate
    Lane 3 : NIH/3T3 (Mouse embyro fibroblast cells) cell lysate
    Lane 4 : RAW264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution

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



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Anti-Lamin B1 antibody [EPR8985(B)] (ab133741) at 1/10000 dilution (purified) + Jurkat (Human T cell leukemia cells from peripheral blood) cell lysate at 10 µg

    Secondary
    Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

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



    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling Lamin B1 with unpurified ab133741 at 1/250.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • All lanes : Anti-Lamin B1 antibody [EPR8985(B)] (ab133741) at 1/10000 dilution

    Lane 1 : Mouse brain lysates
    Lane 2 : Mouse heart lysates
    Lane 3 : Mouse kidney lysates
    Lane 4 : Mouse spleen lysates
    Lane 5 : Rat brain lysates
    Lane 6 : Rat heart lysates
    Lane 7 : Rat spleen lysates
    Lane 8 : C6 (Rat glial tumor cells) whole cell lysates
    Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
    Lane 10 : NIH/3T3 (Mouse embyro fibroblast cells)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

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



    Blocking and Diluting buffer and concentration: 5% NFDM/TBST

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling Lamin B1 with unpurified ab133741 at 1/250.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • ab133741 (purified) at 1/20 immunoprecipitating Lamin B1 in Jurkat cells (Lane 1).  For western blotting, ab133741 was used at 1/1000 dilution and an HRP-conjugated goat anti-rabbit IgG was used as the secondary antibody (1/1500).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

References

This product has been referenced in:

  • Ji XJ  et al. Exendin-4 Exacerbates Burn-Induced Morbidity in Mice by Activation of the Sympathetic Nervous System. Mediators Inflamm 2019:2750528 (2019). Read more (PubMed: 30800001) »
  • Kucharzewska P  et al. NIK-IKK complex interaction controls NF-?B-dependent inflammatory activation of endothelium in response to LTßR ligation. J Cell Sci 132:N/A (2019). Read more (PubMed: 30837284) »
See all 53 Publications for this product

Customer reviews and Q&As

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1-10 of 11 Abreviews

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (hiPSC)
Permeabilization
Yes - 0.1% Triton X
Specification
hiPSC
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1%
Fixative
Methanol

Abcam user community

Verified customer

Submitted Jul 08 2019

Application
Western blot
Sample
Human Cell lysate - nuclear (ARPE-19 cells)
Gel Running Conditions
Reduced Denaturing (4-12% BisTris Gel)
Loading amount
10 µg
Treatment
H2O2 for 10 min
Specification
ARPE-19 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Abcam user community

Verified customer

Submitted May 21 2019

Application
Western blot
Sample
Mouse Tissue lysate - nuclear (mouse RPE nuclear fractions)
Gel Running Conditions
Reduced Denaturing (4-12% BisTris Gel)
Loading amount
10 µg
Treatment
smoke extract injection
Specification
mouse RPE nuclear fractions
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Dr. Lei Wang

Verified customer

Submitted May 21 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Melanoma cell)
Permeabilization
Yes - 0.1% Triton 100
Specification
Melanoma cell
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 1%
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 12 2019

Application
Western blot
Sample
Mouse Cell lysate - whole cell (MEF cells)
Gel Running Conditions
Reduced Denaturing (4-12% Gradient Gel)
Loading amount
50 µg
Specification
MEF cells
Blocking step
Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Aug 01 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (293T cells)
Gel Running Conditions
Reduced Denaturing (4-12% Gradient Gel)
Loading amount
50 µg
Specification
293T cells
Blocking step
Licor Blocking Buffer as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 25°C

Abcam user community

Verified customer

Submitted May 03 2018

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Pig Tissue sections (Intestine, spleen, liver.)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: EDTA pH9
Permeabilization
No
Specification
Intestine, spleen, liver.
Blocking step
Serum as blocking agent for 20 minute(s) · Concentration: 2.5% · Temperature: 25°C
Fixative
Formaldehyde

Victor Bernal

Verified customer

Submitted Apr 23 2018

Application
Western blot
Sample
Mouse Cell lysate - nuclear (mouse macrophage cell line)
Gel Running Conditions
Reduced Denaturing (10% acylamide)
Loading amount
10 µg
Specification
mouse macrophage cell line
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Mar 28 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Dog Cell (Kidney)
Permeabilization
Yes - 0.5% triton x 100
Specification
Kidney
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Fixative
Paraformaldehyde

Mr. Aravindh Subramani

Verified customer

Submitted Apr 18 2016

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Cell lines: SH-SY5Y (neuronal, human) MLP29 (hepat)
Gel Running Conditions
Non-reduced Denaturing (4-12%)
Loading amount
20 µg
Specification
Cell lines: SH-SY5Y (neuronal, human) MLP29 (hepat
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.05% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jan 28 2016

1-10 of 11 Abreviews

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