Anti-Lamin B2抗体[LN43] (ab8983)

概述

  • 产品名称Anti-Lamin B2抗体[LN43]
    参阅全部 Lamin B2 一抗
  • 描述
    小鼠单克隆抗体[LN43] to Lamin B2
  • 特异性This antibody reacts with an epitope located in the C-terminal part of lamin B2. Lamins do not appear to be universally distributed among different cell and tissue types. Please see the listed positive controls for this antibody. Other cell/tissue types have not been tested.
  • 经测试应用适用于: ICC, Flow Cyt, IHC-Fr, WB, IPmore details
  • 种属反应性
    与反应: Mouse, Hamster, Human, Xenopus laevis, Zebrafish
  • 免疫原

    Tissue, cells or virus corresponding to Lamin B2. Immunogen = detergent insoluble fraction of potoroo cell line PtK1.

  • 表位Epitope located in the C-terminal part of lamin B2.
  • 阳性对照
    • IHC-Fr: testicular germ cell tumour sections (see Machiels et al 1997), parabasal cells of skin epidermis (see Broers et al). WB: NCI-H125 nuclear cell lysate Jurkat cells.

性能

应用

Our Abpromise guarantee covers the use of ab8983 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 1/400.
Flow Cyt Use 1µg for 106 cells.

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

IHC-Fr 1/1.

(when used with an avidin-biotinylated HRP complex as detection agent).

WB 1/100 - 1/1000. Detects a band of approximately 65-67 kDa.
IP Use at an assay dependent concentration.

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 LMNB2 are a cause of partial acquired lipodystrophy (APLD) [MIM:608709]. A rare childhood disease characterized by loss of subcutaneous fat from the face and trunk. Fat deposition on the pelvic girdle and lower limbs is normal or excessive. Most frequently, onset between 5 and 15 years of age. Most affected subjects are females and some show no other abnormality, but many develop glomerulonephritis, diabetes mellitus, hyperlipidemia, and complement deficiency. Mental retardation in some cases. APLD is a sporadic disorder of unknown etiology.
  • 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
    • LAMB 2 antibody
    • LAMB2 antibody
    • Lamin-B2 antibody
    • LMN 2 antibody
    • LMN B2 antibody
    • LMN2 antibody
    • LMNB 2 antibody
    • LMNB2 antibody
    • LMNB2_HUMAN antibody
    • MGC2721 antibody
    • RGD1563803 antibody
    see all

Anti-Lamin B2 antibody [LN43] 图像

  •  IF staining of a 9 days old zebrafish embryo.  

  • Immunofluorescence analysis of HeLa cells in telophase, staining Lamin B2 with ab8983.
  • All lanes : Anti-Lamin B2 antibody [LN43] (ab8983) at 1 µg/ml

    Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate
    Lane 3 : U2OS (Human osteosarcoma cell line) Whole Cell Lysate
    Lane 4 : HL60 (Human promyelocytic leukemia cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Observed band size : 75 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 37 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 20 minutes
  • IHC-Fr of human kidney showing nuclear lamina staining in the ductal epithelium.  

  • HeLa cells were fixed in 100% methanol for 6 minutes at -20°C then stained with ab8983 (1/400). This image shows that ab8983 clearly stains the nuclear envelope (green). The DNA is stained with DAPI (blue).

  • Lamin B2 - Nuclear Envelope Marker was immunoprecipitated using 0.5mg Hela whole cell extract, 5µg of Mouse monoclonal to Lamin B2 - Nuclear Envelope Marker (ab8983) and 50µl of protein G magnetic beads (+). No antibody was added to the control (-).
    The antibody was incubated under agitation with Protein G beads for 10min, Hela whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.
    Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab8983.
    Secondary: Goat polyclonal to mouse IgG light chain specific (HRP) at 1/5000 dilution.
    Band: 73kDa: Lamin B2
  • Overlay histogram showing HeLA cells stained with ab8983 (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 (ab8983, 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.

Anti-Lamin B2 antibody [LN43] (ab8983)参考文献

This product has been referenced in:
  • Kuga T  et al. Lamin B2 prevents chromosome instability by ensuring proper mitotic chromosome segregation. Oncogenesis 3:e94 (2014). IP ; Human . Read more (PubMed: 24637494) »
  • Pitcher DS  et al. Nuclear proteasomes carry a constitutive posttranslational modification which derails SDS-PAGE (but not CTAB-PAGE). Biochim Biophys Acta 1844:2222-2228 (2014). Read more (PubMed: 25192768) »

See all 14 Publications for this product

Product Wall

Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (Hela K)
Permeabilization Yes - CSK buffer
Specification Hela K
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: RT°C
Fixative Paraformaldehyde
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提交于 Nov 09 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 24°C
Sample Mouse Cell (Neuronal progenitor cell line)
Specification Neuronal progenitor cell line
Permeabilization Yes - PBS 1% BSA 0,1% Saponine
Fixative Paraformaldehyde
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提交于 Apr 08 2014

Application Western blot
Loading amount 20 µg
Gel Running Conditions Non-reduced Denaturing (precast 4-15%)
Sample Human Cell lysate - whole cell (breast cell line)
Specification breast cell line
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
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提交于 Mar 20 2014

Apologies for the misunderstanding but I thought you wanted a replacement instead of a refund. As requested I have credited the cost of ab8983. Your credit note number is ******.

If you have any further questions on the credit note, please co...

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Thank you for contacting Abcam yesterday.

As we discussed I am sending you a vial of ab97513 as a replacement for ab8983. Your new order/reference number is ********** and you should have it tomorrow morning.

If there is anything el...

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Abcam has not validated the combination of species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Xenopus laevis Cell (nuclei assembled in egg extract)
Specification nuclei assembled in egg extract
Fixative Paraformaldehyde
Permeabilization No
Blocking step BSA as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 4°C
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提交于 Oct 28 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Xenopus laevis Cell lysate - whole cell (meiotic egg extract)
Loading amount 25 µg
Specification meiotic egg extract
Gel Running Conditions Reduced Denaturing (15%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Oct 28 2010

Thank you for your enquiry. Unfortunately, this product was not made from an immunizing peptide, but from a detergent insoluble fraction of potoroo cell line PtK1. Therefore, the immunizing peptide does not exist for this product. I am sorry I ...

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Thank you for your feeback on your experience of ab8983 in your rat brain samples. We have not tested the antibody in this species so it maybe that the protein is truncated or smaller in this species or tissue. Brain contains relatively few nuclei a...

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Thank you for your enquiry and I'm sorry to hear that you are experiencing difficulty with ab8983. At this point I would like to make the following suggestions. Try using nuclear extract rather than whole cell extract, and with the nuclear extract incu...

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