使用敲除细胞株进行验证

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

概述

  • 产品名称Anti-Lamin B1抗体-核Envelope Marker
    参阅全部 Lamin B1 一抗
  • 描述
    兔多克隆抗体to Lamin B1 -核Envelope Marker
  • 特异性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.
  • 经测试应用适用于: ICC/IF, IHC-Fr, WB, IHC-P, IHC - Wholemountmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, Pig, Xenopus laevis, Indian Muntjac
    预测可用于: Chicken, Zebrafish
  • 免疫原

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

    (Peptide available as ab16375.)

  • 阳性对照
    • 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.
  • 常规说明

     

    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.

性能

应用

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.

应用 Ab评论 说明
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

靶标

  • 功能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.
  • 疾病相关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.
  • 序列相似性Belongs to the intermediate filament family.
  • 翻译后修饰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.
  • 细胞定位Nucleus inner membrane.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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 图像

  • 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) ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) 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)参考文献

This product has been referenced in:
  • Cruz OH  et al. Multinucleation and Polykaryon Formation is Promoted by the EhPC4 Transcription Factor in Entamoeba histolytica. Sci Rep 6:19611 (2016). IF . Read more (PubMed: 26792358) »
  • Moreno A  et al. Unreplicated DNA remaining from unperturbed S phases passes through mitosis for resolution in daughter cells. Proc Natl Acad Sci U S A N/A:N/A (2016). WB ; Human . Read more (PubMed: 27516545) »

See all 166 Publications for this product

Product Wall

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
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提交于 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)
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提交于 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)
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提交于 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
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提交于 Nov 04 2016

Application Western blot
Sample Dog Cell lysate - whole cell (Canine Kidney Epithelial MDCK cells)
Gel Running Conditions Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount 500000 cells
Specification Canine Kidney Epithelial MDCK 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
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提交于 Oct 27 2016

Application Immunohistochemistry (Frozen sections)
Sample Human Tissue sections (Mammary epithelial cell monolayer)
Permeabilization Yes - Triton
Specification Mammary epithelial cell monolayer
Blocking step Serum as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative Formaldehyde
Username

Dr. Claire Robertson

Verified customer

提交于 Sep 09 2016

Application Western blot
Sample African Green Monkey Cell lysate - whole cell (VERO E6)
Gel Running Conditions Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount 50000 cells
Specification VERO E6
Blocking step Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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提交于 Aug 31 2016

Application Western blot
Sample Human Cell lysate - whole cell (A549 (human lung epithelium cells))
Gel Running Conditions Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount 50000 cells
Treatment 200 units/mL human IFN beta
Specification A549 (human lung epithelium 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
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提交于 Aug 22 2016

Application Western blot
Sample Hamster Cell lysate - whole cell (BHK-21 (Kidney Fibroblasts))
Gel Running Conditions Reduced Denaturing (4-12% Bolt Bis-Tris Plus gel)
Loading amount 80000 cells
Specification BHK-21 (Kidney Fibroblasts)
Blocking step Li-Cor Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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提交于 Aug 22 2016

Application Western blot
Sample Human Cell lysate - nuclear (HEK293)
Gel Running Conditions Reduced Denaturing
Loading amount 10 µg
Specification HEK293
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Dr. Alexander Garvin

Verified customer

提交于 Jul 05 2016

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