使用敲除细胞株进行验证

Anti-Lamin B1抗体[119D5-F1] (ab8982)

概述

  • 产品名称
    Anti-Lamin B1抗体[119D5-F1]
    参阅全部 Lamin B1 一抗
  • 描述
    小鼠单克隆抗体[119D5-F1] to Lamin B1
  • 特异性
    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.
  • 经测试应用
    适用于: WB, Dot blot, ELISA, Flow Cyt, ICC, ICC/IF, IHC-Frmore details
  • 种属反应性
    与反应: Mouse, Rat, Sheep, Rabbit, Cow, Dog, Human
  • 免疫原

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

  • 表位
    C-terminal to residue 231.
  • 阳性对照
    • ICC/IF: Wildtype HAP1 cells.

性能

应用

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.

应用 Ab评论 说明
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.

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

图片

  • 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
  • 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
    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

    Image courtesy of an anonymous Abreview.

    See Abreview

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

文献

This product has been referenced in:
  • Li F  et al. Enhancement of early cardiac differentiation of dedifferentiated fat cells by dimethyloxalylglycine via notch signaling pathway. Am J Transl Res 8:4791-4801 (2016). WB ; Rat . Read more (PubMed: 27904680) »
  • Sun X  et al. Disruption of endothelial cell mitochondrial bioenergetics in lambs with increased pulmonary blood flow. Antioxid Redox Signal 18:1739-52 (2013). Read more (PubMed: 23244702) »

See all 15 Publications for this product

客户评价及客户问答

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
Username

Juliana Schneider

Verified customer

提交于 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
Username

Abcam user community

Verified customer

提交于 Oct 27 2014

Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this h...

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Have a lovely holiday and please ask your colleague to confirm if you wish to get a replacement and when.

I look forward to hearing from you soon.

Thank you for getting back to me and for providing some further details.

I would like to reassure you that our Abpromise applies to your complaint since you purchased this product within the guarantee period. This means that in the event that...

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Thank you for your enquiry regarding ab8982 and for taking the time to provide some useful details of the experiments. I am very sorry to hear that your customer is having problems with this antibody.

A) Image: Both images labelled as PPAR. I...

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I am sorry to hear that you have been experiencing problems using this product in the application that you wish.

All our customer feedback, including complaints are monitored weekly by our in house technical support team. If a product is at f...

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Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

提交于 Feb 22 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Abcam user community

Verified customer

提交于 Dec 30 2010

I'm sorry to hear you are having a problem with ab8982. This antibody has been purchased many times in the past few months without any reported problems, so I am fairly confident that we can modify your protocol and get the antibody to work for you....

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

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