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  1. Link

    lamin-a-antibody-133a2-ab8980.pdf

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Tags & Cell Markers Subcellular Markers Nucleus Nuclear Envelope
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Validated using a knockout cell line

Anti-Lamin A antibody [133A2] (ab8980)

  • Datasheet
  • SDS
Reviews (17)Q&A (20)References (77)

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Western blot - Anti-Lamin A antibody [133A2] (ab8980)
  • Immunocytochemistry/ Immunofluorescence - Anti-Lamin A antibody [133A2] (ab8980)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A antibody [133A2] (ab8980)
  • Flow Cytometry - Anti-Lamin A antibody [133A2] (ab8980)
  • Immunocytochemistry/ Immunofluorescence - Anti-Lamin A antibody [133A2] (ab8980)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A antibody [133A2] (ab8980)
  • Immunohistochemistry (Frozen sections) - Anti-Lamin A antibody [133A2] (ab8980)
  • Immunocytochemistry/ Immunofluorescence - Anti-Lamin A antibody [133A2] (ab8980)

Key features and details

  • Mouse monoclonal [133A2] to Lamin A
  • Suitable for: Flow Cyt, IHC-Fr, WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human
  • Isotype: IgG3

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Overview

  • Product name

    Anti-Lamin A antibody [133A2]
    See all Lamin A primary antibodies
  • Description

    Mouse monoclonal [133A2] to Lamin A
  • Host species

    Mouse
  • Specificity

    Reacts against lamin A exclusively (the antibody was raised against the carboxy-terminus of 98 amino acids present in lamin A and absent from lamin C. As a result, this antibody recognizes lamin A but not lamin C; See Machiels et al 1997). Lamins do not appear to be universally distributed among different cell and tissue types. ab8980 has been tested in testis parenchyma and testicular germ cell tumours (See Machiels et al 1997). Other cell/tissue types have not been tested.
  • Tested applications

    Suitable for: Flow Cyt, IHC-Fr, WB, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Recombinant fragment corresponding to Human Lamin A (C terminal).
    Database link: P02545

  • Epitope

    Recognizes an epitope located between residues 598-611 of lamin A.
  • Positive control

    • Mouse heart normal tissue lysate - total protein (ab30291) can be used as a positive control in WB. This antibody gave a positive signal in the following cell types: HeLa WB: Wild-type HAP1 whole cell lysate; HeLa whole cell lysate
  • General notes

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.09% Sodium azide
    Constituent: 99% PBS

    This IgG3 antibody has to be purified from the culture medium by ion-exchange chromatography, therefore it will contain remnants of bovine serum albumin.
  • Concentration information loading...
  • Purity

    Ion Exchange Chromatography
  • Clonality

    Monoclonal
  • Clone number

    133A2
  • Myeloma

    P3x63-Ag8.653
  • Isotype

    IgG3
  • Light chain type

    kappa
  • Research areas

    • Tags & Cell Markers
    • Subcellular Markers
    • Nucleus
    • Nuclear Envelope
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Intermediate Filaments
    • Class V
    • Lamins

Associated products

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

    • Mouse IgG3, kappa monoclonal [MG3-35] - Isotype Control (ab18394)
  • Positive Controls

    • Mouse heart normal tissue lysate - total protein (ab30291)
  • Recombinant Protein

    • Recombinant Human Lamin A protein (ab83472)
  • Related Products

    • 10x Citrate Buffer pH 6.0 (ab64214)
    • Normal Goat Serum (ab7481)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab8980 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
Flow Cyt
1/100 - 1/200.

ab18392 - Mouse monoclonal IgG3, is suitable for use as an isotype control with this antibody.

IHC-Fr (1)
1/100 - 1/200.
WB (6)
1/100 - 1/1000. Predicted molecular weight: 74 kDa.

PMID: 9274531

IHC-P (4)
1/100 - 1/200.
ICC/IF (6)
Use a concentration of 1 µg/ml.
Notes
Flow Cyt
1/100 - 1/200.

ab18392 - Mouse monoclonal IgG3, is suitable for use as an isotype control with this antibody.

IHC-Fr
1/100 - 1/200.
WB
1/100 - 1/1000. Predicted molecular weight: 74 kDa.

PMID: 9274531

IHC-P
1/100 - 1/200.
ICC/IF
Use a concentration of 1 µg/ml.

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. Lamin A and C are present in equal amounts in the lamina of mammals. Play an important role in nuclear assembly, chromatin organization, nuclear membrane and telomere dynamics.
    Prelamin-A/C can accelerate smooth muscle cell senescence. It acts to disrupt mitosis and induce DNA damage in vascular smooth muscle cells (VSMCs), leading to mitotic failure, genomic instability, and premature senescence.
  • Tissue specificity

    In the arteries, prelamin-A/C accumulation is not observed in young healthy vessels but is prevalent in medial vascular smooth muscle celle (VSMCs) from aged individuals and in atherosclerotic lesions, where it often colocalizes with senescent and degenerate VSMCs. Prelamin-A/C expression increases with age and disease. In normal aging, the accumulation of prelamin-A/C is caused in part by the down-regulation of ZMPSTE24/FACE1 in response to oxidative stress.
  • Involvement in disease

    Defects in LMNA are the cause of Emery-Dreifuss muscular dystrophy type 2 (EDMD2) [MIM:181350]. A degenerative myopathy characterized by weakness and atrophy of muscle without involvement of the nervous system, early contractures of the elbows, Achilles tendons and spine, and cardiomyopathy associated with cardiac conduction defects.
    Defects in LMNA are the cause of cardiomyopathy dilated type 1A (CMD1A) [MIM:115200]. Dilated cardiomyopathy is a disorder characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia. Patients are at risk of premature death.
    Defects in LMNA are the cause of familial partial lipodystrophy type 2 (FPLD2) [MIM:151660]; also known as familial partial lipodystrophy Dunnigan type. A disorder characterized by the loss of subcutaneous adipose tissue in the lower parts of the body (limbs, buttocks, trunk). It is accompanied by an accumulation of adipose tissue in the face and neck causing a double chin, fat neck, or cushingoid appearance. Adipose tissue may also accumulate in the axillae, back, labia majora, and intraabdominal region. Affected patients are insulin-resistant and may develop glucose intolerance and diabetes mellitus after age 20 years, hypertriglyceridemia, and low levels of high density lipoprotein cholesterol.
    Defects in LMNA are the cause of limb-girdle muscular dystrophy type 1B (LGMD1B) [MIM:159001]. LGMD1B is an autosomal dominant degenerative myopathy with age-related atrioventricular cardiac conduction disturbances, dilated cardiomyopathy, and the absence of early contractures. LGMD1B is characterized by slowly progressive skeletal muscle weakness of the hip and shoulder girdles. Muscle biopsy shows mild dystrophic changes.
    Defects in LMNA are the cause of Charcot-Marie-Tooth disease type 2B1 (CMT2B1) [MIM:605588]. CMT2B1 is a form of Charcot-Marie-Tooth disease, the most common inherited disorder of the peripheral nervous system. Charcot-Marie-Tooth disease is classified in two main groups on the basis of electrophysiologic properties and histopathology: primary peripheral demyelinating neuropathy or CMT1, and primary peripheral axonal neuropathy or CMT2. Neuropathies of the CMT2 group are characterized by signs of axonal regeneration in the absence of obvious myelin alterations, normal or slightly reduced nerve conduction velocities, and progressive distal muscle weakness and atrophy. CMT2B1 inheritance is autosomal recessive.
    Defects in LMNA are the cause of Hutchinson-Gilford progeria syndrome (HGPS) [MIM:176670]. HGPS is a rare genetic disorder characterized by features reminiscent of marked premature aging. Note=HGPS is caused by the toxic accumulation of a mutant form of lamin-A/C. This mutant protein, called progerin, acts to deregulate mitosis and DNA damage signaling, leading to premature cell death and senescence. Progerin lacks the conserved ZMPSTE24/FACE1 cleavage site and therefore remains permanently farnesylated. Thus, although it can enter the nucleus and associate with the nuclear envelope, it cannot incorporate normally into the nuclear lamina.
    Defects in LMNA are the cause of cardiomyopathy dilated with hypergonadotropic hypogonadism (CMDHH) [MIM:212112]. A disorder characterized by the association of genital anomalies, hypergonadotropic hypogonadism and dilated cardiomyopathy. Patients can present other variable clinical manifestations including mental retardation, skeletal anomalies, scleroderma-like skin, graying and thinning of hair, osteoporosis. Dilated cardiomyopathy is characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia.
    Defects in LMNA are the cause of mandibuloacral dysplasia with type A lipodystrophy (MADA) [MIM:248370]. A disorder characterized by mandibular and clavicular hypoplasia, acroosteolysis, delayed closure of the cranial suture, progeroide appearance, partial alopecia, soft tissue calcinosis, joint contractures, and partial lipodystrophy with loss of subcutaneous fat from the extremities. Adipose tissue in the face, neck and trunk is normal or increased.
    Defects in LMNA are a cause of lethal tight skin contracture syndrome (LTSCS) [MIM:275210]; also known as restrictive dermopathy (RD). Lethal tight skin contracture syndrome is a rare disorder mainly characterized by intrauterine growth retardation, tight and rigid skin with erosions, prominent superficial vasculature and epidermal hyperkeratosis, facial features (small mouth, small pinched nose and micrognathia), sparse/absent eyelashes and eyebrows, mineralization defects of the skull, thin dysplastic clavicles, pulmonary hypoplasia, multiple joint contractures and an early neonatal lethal course. Liveborn children usually die within the first week of life. The overall prevalence of consanguineous cases suggested an autosomal recessive inheritance.
    Defects in LMNA are the cause of heart-hand syndrome Slovenian type (HHS-Slovenian) [MIM:610140]. Heart-hand syndrome (HHS) is a clinically and genetically heterogeneous disorder characterized by the co-occurrence of a congenital cardiac disease and limb malformations.
    Defects in LMNA are the cause of muscular dystrophy congenital LMNA-related (CMD-LMNA) [MIM:613205]. It is a form of congenital muscular dystrophy. Patients present at birth, or within the first few months of life, with hypotonia, muscle weakness and often with joint contractures.
  • Sequence similarities

    Belongs to the intermediate filament family.
  • Post-translational
    modifications

    Increased phosphorylation of the lamins occurs before envelope disintegration and probably plays a role in regulating lamin associations.
    Proteolytic cleavage of the C-terminal of 18 residues of prelamin-A/C results in the production of lamin-A/C. The prelamin-A/C maturation pathway includes farnesylation of CAAX motif, ZMPSTE24/FACE1 mediated cleavage of the last three amino acids, methylation of the C-terminal cysteine and endoproteolytic removal of the last 15 C-terminal amino acids. Proteolytic cleavage requires prior farnesylation and methylation, and absence of these blocks cleavage.
    Sumoylation is necessary for the localization to the nuclear envelope.
    Farnesylation of prelamin-A/C facilitates nuclear envelope targeting.
  • Cellular localization

    Nucleus. Nucleus envelope. Farnesylation of prelamin-A/C facilitates nuclear envelope targeting and subsequent cleaveage by ZMPSTE24/FACE1 to remove the farnesyl group produces mature lamin-A/C, which can then be inserted into the nuclear lamina. EMD is required for proper localization of non-farnesylated prelamin-A/C.
  • Target information above from: UniProt accession P02545 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 4000 Human
    • Entrez Gene: 16905 Mouse
    • Omim: 150330 Human
    • SwissProt: P02545 Human
    • SwissProt: P48678 Mouse
    • Unigene: 594444 Human
    • Unigene: 243014 Mouse
    • Unigene: 471227 Mouse
    • Alternative names

      • 70 kDa lamin antibody
      • CDDC antibody
      • EMD2 antibody
      • FPL antibody
      • FPLD antibody
      • HGPS antibody
      • IDC antibody
      • LAMIN A antibody
      • lamin A/C antibody
      • LAMIN C antibody
      • Lamin-A/C antibody
      • LDP1 antibody
      • LFP antibody
      • LMN 1 antibody
      • LMN A antibody
      • LMN C antibody
      • LMNA antibody
      • LMNA_HUMAN antibody
      • LMNC antibody
      • PRO1 antibody
      • Renal carcinoma antigen NY-REN-32 antibody
      see all

    Images

    • Western blot - Anti-Lamin A antibody [133A2] (ab8980)
      Western blot - Anti-Lamin A antibody [133A2] (ab8980)
      All lanes : Anti-Lamin A antibody [133A2] (ab8980) at 1/100 dilution

      Lane 1 : Wild-type HAP1 whole cell lysate
      Lane 2 : LMNA knockout HAP1 whole cell lysate
      Lane 3 : HeLa whole cell lysate
      Lane 4 : HepG2 whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Predicted band size: 74 kDa
      Observed band size: 74 kDa



      Lanes 1 - 4: Merged signal (red and green). Green - ab8980 observed at 74 kDa. Red - loading control, ab181602, observed at 37 kDa.

      ab8980 was shown to recognize  in wild-type HAP1 cells as signal was lost at the expected MW in LMNA knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and LMNA knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3pc Milk. Ab8980 and ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1/100 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye® 800CW) preabsorbed ab216772 and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    • Immunocytochemistry/ Immunofluorescence - Anti-Lamin A antibody [133A2] (ab8980)
      Immunocytochemistry/ Immunofluorescence - Anti-Lamin A antibody [133A2] (ab8980)

      ICC/IF image of ab8980 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum (ab7481) / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab8980 at 1µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (ab96879) IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A antibody [133A2] (ab8980)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A antibody [133A2] (ab8980)This image is courtesy of an Abreview submitted by Mr Carl Hobbs

      ab8980 staining of Lamin A in adult mouse liver sections. Formaldehyde-fixed paraffin-embedded sections of mouse liver tissue were incubated with ab8980 (1/2500) fo 2 hours. Antigen retrieval was performed by heat induction in citrate buffer pH 6.0 (ab64214). Please see accompanying abreview for additional information.

      See Abreview

    • Flow Cytometry - Anti-Lamin A antibody [133A2] (ab8980)
      Flow Cytometry - Anti-Lamin A antibody [133A2] (ab8980)
      Overlay histogram showing HeLa cells stained with ab8980 (red line). The cells were fixed with 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 / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab8980, 1/100 dilution) 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 IgG3 [MG3-35] (2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.

    • Immunocytochemistry/ Immunofluorescence - Anti-Lamin A antibody [133A2] (ab8980)
      Immunocytochemistry/ Immunofluorescence - Anti-Lamin A antibody [133A2] (ab8980)
      These images show HeLa cells stained with Laminin A antibody - ab8980 (green) and with DAPI (blue). The pictures were kindly supplied as part of the review submitted by Dr Josef Gotzmann at Medical University of Vienna, Austria.
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A antibody [133A2] (ab8980)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin A antibody [133A2] (ab8980)

      Immunohistochemistry on paraffin section of human colon.

    • Immunohistochemistry (Frozen sections) - Anti-Lamin A antibody [133A2] (ab8980)
      Immunohistochemistry (Frozen sections) - Anti-Lamin A antibody [133A2] (ab8980)

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

    • Immunocytochemistry/ Immunofluorescence - Anti-Lamin A antibody [133A2] (ab8980)
      Immunocytochemistry/ Immunofluorescence - Anti-Lamin A antibody [133A2] (ab8980)
      Immunocytochemical staining of fiboblasts showing nuclear lamina

    Protocols

    • Recommended WB protocol for lamin proteins
    • Recommended IHC-Fr protocol for lamin proteins

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (77)

    Publishing research using ab8980? Please let us know so that we can cite the reference in this datasheet.

    ab8980 has been referenced in 77 publications.

    • Guo H  et al. Carthamin yellow improves cerebral ischemia-reperfusion injury by attenuating inflammation and ferroptosis in rats. Int J Mol Med 47:1 (2021). PubMed: 33576458
    • Antmen E  et al. Micropatterned Surfaces Expose the Coupling between Actin Cytoskeleton-Lamin/Nesprin and Nuclear Deformability of Breast Cancer Cells with Different Malignancies. Adv Biol (Weinh) 5:e2000048 (2021). PubMed: 33724728
    • Zhang S  et al. Neuroprotective mechanisms of e-viniferin in a rotenone-induced cell model of Parkinson's disease: significance of SIRT3-mediated FOXO3 deacetylation. Neural Regen Res 15:2143-2153 (2020). PubMed: 32394973
    • Yang Y  et al. Lack of sphingomyelin synthase 2 reduces cerebral ischemia/reperfusion injury by inhibiting microglial inflammation in mice. Exp Ther Med 20:241 (2020). PubMed: 33178339
    • Fan JR  et al. Lamin A-mediated nuclear lamina integrity is required for proper ciliogenesis. EMBO Rep 21:e49680 (2020). PubMed: 32815283
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 37 Abreviews or Q&A

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Lamin A antibody [133A2]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (MEFs)
    Permeabilization
    Yes - PBST
    Specification
    MEFs
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More

    Mr. Yuanxin Yang

    Verified customer

    Submitted Dec 24 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Lamin A antibody [133A2]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Mouse Cell (MEF cells)
    Permeabilization
    Yes - 0.2% Triton X100
    Specification
    MEF cells
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Nov 20 2017

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Lamin A antibody [133A2]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (MCF10A)
    Permeabilization
    Yes - 0.2% Triton x100
    Specification
    MCF10A
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0.1% · Temperature: 25°C
    Fixative
    Paraformaldehyde
    Read More

    Abcam user community

    Verified customer

    Submitted Mar 30 2017

    Western blot abreview for Anti-Lamin A [133A2] antibody

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Loading amount
    15 µg
    Gel Running Conditions
    Non-reduced Denaturing (8%)
    Sample
    Mouse Cell lysate - whole cell (Mice brain tissue)
    Specification
    Mice brain tissue
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 24°C
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 15 2014

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Lamin A [133A2] antibody

    Good
    Abreviews
    Abreviews
    abreview image
    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
    Read More

    Abcam user community

    Verified customer

    Submitted Apr 03 2014

    Question

    I wanted to ask a question about the antibody: ‘Anti-Lamin A antibody [133A2] (ab8980)’. Does this antibody also stain positive for progerin, and has this been tested by you or any of your customers? Since it is monoclonal we need to be sure whether it also stains for progerin. Thanks in advance for your answer.

    Read More

    Abcam community

    Verified customer

    Asked on Nov 26 2014

    Answer

    We have not previously tested whether this antibody stains progerin.

    The epitope of ab8980 corresponds to amino acids 598-611.According to uniprot (http://www.uniprot.org/uniprot/P02545), progerin lacks amino acids 607-656.

    There is therefore 62% of the epitope amino acids present in progerin so it could react however we cannot guarantee this.

    If you would like an anti-lamin A antibody which will also detect progerin, I could recommend Anti-Lamin A antibody (ab26300) whose immunogen corresponds to amino acids 570 to 587 which are also present in progerin.

    Read More

    Elisa Thomas

    Abcam Scientific Support

    Answered on Nov 26 2014

    Question

    I will test my blot with an alpha-tubulin antibody this week or the next week .

    I come back toward you when I will have new results

    Best regards

    Read More

    Abcam community

    Verified customer

    Asked on Oct 02 2012

    Answer

    Thanks! I will look forward to hearing from you soon.

    Read More

    Abcam Scientific Support

    Answered on Oct 02 2012

    Question

    Thank you for your response.

    I used Subcellular Protein Fractionation Kit for Cultured Cells 78840 from Pierce.

    They said that the kit is compatible with mammalian cultured cells.

    So I can used for mouse samples. I have not used theses mouse fractions with an other antibody.

    In my blot with lysates from mouse , I have proteins from extraction with RIPA-N and with fractionned extraction. And no band .

    So I don’t know what I should do.

    What advice would you give me?

    Thank you in advance.

    Best regards

    Read More

    Abcam community

    Verified customer

    Asked on Oct 01 2012

    Answer

    Thank you for your email.

    "In my blot with lysates from mouse , I have proteins from extraction with RIPA-N and with fractionned extraction. And no band." Does this mean you have performed cell lysis with RIPA buffer also and there was no band?

    Do you have other antibodies available e.g. anti actin or beta actin, anti GAPDH etc that can be used to against same lysates. If these antibodies shows positive results then we can assume that lysates are alright and the problem is due to antibody not the kit.

    Thanks! I will be looking forwardto hearing from you soon.

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    Abcam Scientific Support

    Answered on Oct 01 2012

    Question

    Product code: 8980
    Lot number: GR68178-2
    Inquiry: I have bought an anti-Lamin A antibody. it functions in western blot with human cellular cell lines (K562, SET2) I have but no murine (FDCP1, Baf3, EML). I don't know why, I made a fragmented extraction protein : nuclear, bound-chromatin with a kit from Pierce. So I know that I have well nuclear proteins. More I would like know if you have a method to eluate quickly nuclear proteins and bound-chromatin proteins. Thank you. Best regards

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    Abcam community

    Verified customer

    Asked on Oct 01 2012

    Answer

    Thank you for your email. I am sorry to hear that you have been experiencing problems with this antibody.

    The antibody has been characterized with mouse lysates and found to be specific to mouse protein, which is why we are selling this as mouse and human specific product. As the antibody is working well with human lysates so let's keep aside the quality of this, which seems OK. So the problem might be with the fractions prepared from KIT. Could you tell us the catalogue number of Pierce kit used. Is this kit suitable for mouse samples? Have you done any troubleshooting or used any other antibody than ab8980 with same fractions?

    I would suggest preparing mouse cell line lysates without fractions i.e. simple WB using RIPA or other lysis buffer and then checking the results. If antibody fails then antibody is damaged if not then protein extraction protocol might need further updates.

    Please let me know of your thoughts. I hope these suggestions would be helpful.

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    Abcam Scientific Support

    Answered on Oct 01 2012

    Question

    Hi, Could you recommend a good antibody that I could use as loading control for western blot on cell membrane fractions? Thanks

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    Abcam community

    Verified customer

    Asked on Mar 30 2012

    Answer

    Thank you for your enquiry.

    Below is a selection of antibodies that are suitable for detecting the plasma membrane and nuclear envelope. They all work in western blot and I would review the datasheets to confirm which species they are tested against.

    Plasma membrane
    ab4645 Anti-Pma1 antibody [40B7]
    www.abcam.com/4645

    ab7671 Anti-alpha 1 Sodium Potassium ATPase antibody [464.6]
    www.abcam.com/ab7671

    ab16505 Anti-pan Cadherin antibody
    www.abcam.com/ab16505

    ab22744 Anti-pan Cadherin antibody [mAbcam22744]
    www.abcam.com/ab22744

    Nuclear envelope
    ab8980 Anti-Lamin A antibody [133A2]
    www.abcam.com/ab8980

    ab8984 Anti-Lamin A + C antibody [131C3]
    www.abcam.com/ab8984

    ab16048 Anti-Lamin B1 antibody
    www.abcam.com/ab16048

    I hope this is useful. Just let me know if you have further questions.

    Read More

    Abcam Scientific Support

    Answered on Mar 30 2012

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