Anti-Lamin B1 抗体 - Nuclear Envelope Marker (ab16048)

製品の概要

  • 製品名
    Anti-Lamin B1 antibody - Nuclear Envelope Marker
    Lamin B1 一次抗体 製品一覧
  • 製品の詳細
    Rabbit polyclonal to Lamin B1 - Nuclear Envelope Marker
  • 特異性
    Lamins do not appear to be universally distributed among different cell and tissue types. ab16048 has been shown to react with HeLa cells/lysates in Western blot and ICC. Other cell/tissue types have not been tested.
  • アプリケーション
    適用あり: ICC/IF, IHC-Fr, WB, IHC-P, IHC - Wholemountmore details
  • 種交差性
    交差種: Mouse, Rat, Human, Pig, Xenopus laevis, Indian muntjac
    交差が予測される動物種: Chicken, Zebrafish
  • 免疫原

    Synthetic peptide conjugated to KLH derived from within residues 400 - 500 of Mouse Lamin B1.

    (Peptide available as ab16375.)

  • ポジティブ・コントロール
    • This antibody gave a positive signal in the following whole cell lysates: HeLa. This antibody gave a positive signal in the following Methanol fixed cell lines: HeLa. This antibody gave a positive signal in the following FFPE tissue: Human normal liver.
  • 特記事項

    Lamin B1 and Lamin B antibodies are extremely useful as nuclear loading controls for use with nuclear extracts. When using Lamin B1 antibodies as nuclear loading controls, be aware that in apoptotic cells Lamin B1 is cleaved (Kottke TJ et al.). Lamin B1 will also be removed from a nuclear prep if the nuclear membranes are spun out. This antibody was designed to be a nuclear loading control however it has not yet been tested in appropriate lysates.

製品の特性

アプリケーション

Our Abpromise guarantee covers the use of ab16048 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

アプリケーション Abreviews 特記事項
ICC/IF Use a concentration of 1 µg/ml.
IHC-Fr Use at an assay dependent concentration.
WB Use a concentration of 0.1 µg/ml. Detects a band of approximately 68 kDa (predicted molecular weight: 66 kDa).
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC - Wholemount Use at an assay dependent concentration. PubMed: 25368174

ターゲット情報

  • 機能
    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.
  • 関連疾患
    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.
  • 配列類似性
    Belongs to the intermediate filament family.
  • 翻訳後修飾
    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.
  • 細胞内局在
    Nucleus inner membrane.
  • Information by UniProt
  • 参照データベース
  • 別名
    • 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

Anti-Lamin B1 antibody - Nuclear Envelope Marker 画像



  • Predicted band size : 66 kDa

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: empty lane
    Lane 3: LMNB1 whole cell lysate (20 µg)
    Lane 4: empty lane
    Lanes 1 - 4: Merged signal (red and green). Green - ab16048 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab16048 was shown to specifically recognize Lamin B1 when LMNB1 (Lamin B1) knockout samples were used, along with additional cross-reactive bands. Wild-type and knockout samples were subjected to SDS-PAGE. Ab16048 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 0.1 μg per mL 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.

  • Human and mouse cells stained with ab16048 (1/500). The cells were fixed and permeabilized in 4% formaldehyde, 0.2% Tritonm X100 for 10 minutes at room temperature, then washed 3x in PBS.

    A: HeLa cells + ab16048 (green)
    B: HeLa cells counterstained with DAPI (blue)
    C: 3T3 cells + ab16048 (green)
    D: 3T3 cells counterstained with DAPI (blue)



  • Performed under reducing conditions.

    Predicted band size : 66 kDa
    Observed band size : 68 kDa (why is the actual band size different from the predicted?)

    Lane 1: Wild-type HAP1 nuclear lysate (10 µg)
    Lane 2: Lamin B1 knockout HAP1 nuclear lysate (10 µg)

    Lanes 1 and 2: Green signal from target - ab16048 observed at 68 kDa. Red signal from loading control - ab10799 observed at 18 kDa.

    ab16048 was shown to specifically react with lamin B1 when lamin B1 knockout samples were used. Wild-type and lamin B1 knockout samples were subjected to SDS-PAGE. ab16048 and ab10799 (loading control to histone H3 at 0.1µg/mL) were both incubated overnight at 4°C. 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 1/10,000 dilution for 1 h at room temperature before imaging.

  • IHC image of Lamin B1 staining in Human normal Liver formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab16048, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    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 - Nuclear Envelope Marker (ab16048) at 1/1000 dilution

    Lane 1 : Hela whole cell lysate
    Lane 2 : Hela whole cell lysate with Mouse Lamin B1 peptide (ab16375) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    Alexa fluor goat polyclonal to Rabbit IgG at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 66 kDa
    Observed band size : 68-70 kDa (why is the actual band size different from the predicted?)
  • ICC/IF image of ab16048 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab16048, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).

  • Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048) at 1/1000 dilution + Pancreatic cell line - whole cell lysate at 20 µg

    Secondary
    HRP conjugated goat anti-rabbit antibody at 1/2000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 66 kDa
    Observed band size : 68 kDa (why is the actual band size different from the predicted?)


    Exposure time : 30 seconds

    This image is courtesy of an anonymous Abreview

    See Abreview

  • ab16048 staining Lamin B1 in human infantile fibromatosis tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with formaldehyde and blocked with 1% FBS/BSA for 3 hours at room temperature; antigen retrieval was by heat mediation in Tris pH9. Samples were incubated with primary antibody (1/100 in TBS + 1% BSA + 1% FBS) for 16 hours. An undiluted HRP-conjugated goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

  • ab16048 staining Lamin B1 in Pig PAE cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with formaldehyde and blocked with 3% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/100 in PBS + 1% BSA) for 1 hour. An Alexa Fluor® 488-conjugated donkey anti-rabbit IgG polyclonal (1/500) was used as the secondary antibody.

    See Abreview

  • All lanes : Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048) at 1/1000 dilution

    Lane 1 : Pig PAE whole cell lysate
    Lane 2 : Pig PAE whole cell lysate

    Lysates/proteins at 50 µg per lane.

    Secondary
    HRP-conjugated goat anti-rabbit IgG polyclonal at 1/2000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 66 kDa
    Observed band size : 65 kDa (why is the actual band size different from the predicted?)


    Exposure time : 3 minutes

    This image is courtesy of an anonymous Abreview

    See Abreview

  • Human and mouse cells stained with ab16048 (1/500). The cells were fixed in 100% methanol for 6 minutes at -20°C, then washed once in PBS.

    A: HeLa cells + ab16048 (green)
    B: HeLa cells counterstained with DAPI (blue)
    C: 3T3 cells + ab16048 (green)
    D: 3T3 cells counterstained with DAPI (blue)

     

Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048) 使用論文

This product has been referenced in:
  • Ranade D  et al. Chromosomal aneuploidies induced upon Lamin B2 depletion are mislocalized in the interphase nucleus. Chromosoma 126:223-244 (2017). WB, ICC/IF ; Human . Read more (PubMed: 26921073) »
  • Vinyoles M  et al. Activation of CK1? by PP2A/PR61? is required for the initiation of Wnt signaling. Oncogene 36:429-438 (2017). IP ; Human . Read more (PubMed: 27321178) »

See all 309 Publications for this product

Product Wall

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse Derived Breast Cancer Cell Line)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
30 µg
Specification
Mouse Derived Breast Cancer Cell Line
Blocking step
Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 26°C
Username

Abcam user community

Verified customer

投稿 Oct 09 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (smooth muscle cells)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
30 µg
Specification
smooth muscle cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 May 31 2017

Application
Western blot
Sample
Human Cell lysate - nuclear (Vascular smooth muscle cell)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Specification
Vascular smooth muscle cell
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

投稿 May 24 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - nuclear (SKOv3 cells)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Loading amount
20 µg
Specification
SKOv3 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

投稿 Mar 28 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - other (nuclear and cytoplasmic fraction from skin keratin)
Gel Running Conditions
Reduced Denaturing (any Kd BioRad)
Loading amount
20 µg
Specification
nuclear and cytoplasmic fraction from skin keratin
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Mr. Julien Coutier

Verified customer

投稿 Feb 07 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Chinese hamster Cell lysate - whole cell (Chinese hamster ovary (CHO) cells)
Gel Running Conditions
Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel (MES buffer))
Loading amount
50000 cells
Specification
Chinese hamster ovary (CHO) cells
Blocking step
Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

投稿 Dec 31 2016

Application
Western blot
Sample
Cow Cell lysate - nuclear (bovine macrophages)
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Loading amount
5 µg
Treatment
cytosol and nuclear fraction
Specification
bovine macrophages
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 18°C
Username

Abcam user community

Verified customer

投稿 Nov 17 2016

Application
Immunoprecipitation
Sample
Mouse Cell lysate - whole cell (Mouse embryonic fibroblasts (MEFs))
Total protein in input
1e+007 cells
Immuno-precipitation step
Other - Protein G Dynabeads
Specification
Mouse embryonic fibroblasts (MEFs)
Username

Abcam user community

Verified customer

投稿 Nov 04 2016

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (A549 (human lung epithelium cells))
Total protein in input
1e+007 cells
Immuno-precipitation step
Other - Protein G Dynabeads
Specification
A549 (human lung epithelium cells)
Username

Abcam user community

Verified customer

投稿 Nov 04 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Dog Cell lysate - whole cell (Canine Kidney Epithelial MDCK cells)
Total protein in input
5e+006 cells
Immuno-precipitation step
Other - Protein G Dynabeads
Specification
Canine Kidney Epithelial MDCK cells
Username

Abcam user community

Verified customer

投稿 Nov 04 2016

1-10 of 99 Abreviews or Q&A

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

登録