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Cell Biology Apoptosis Nucleus Lamins
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Validated using a knockout cell line

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

  • Datasheet
  • SDS
Reviews (96)Q&A (16)References (625)

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Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
  • Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
  • Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
  • Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
  • Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
  • Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
  • Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)

Key features and details

  • Rabbit polyclonal to Lamin B1 - Nuclear Envelope Marker
  • Suitable for: ICC/IF, WB, IHC-P
  • Knockout validated
  • Reacts with: Mouse, Human
  • Isotype: IgG

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Overview

  • Product name

    Anti-Lamin B1 antibody - Nuclear Envelope Marker
    See all Lamin B1 primary antibodies
  • Description

    Rabbit polyclonal to Lamin B1 - Nuclear Envelope Marker
  • Host species

    Rabbit
  • Tested applications

    Suitable for: ICC/IF, WB, IHC-Pmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat, Chicken, Pig, Xenopus laevis, Indian muntjac, Zebrafish
  • Immunogen

    Synthetic peptide corresponding to Mouse Lamin B1 aa 400-500 (internal sequence) conjugated to keyhole limpet haemocyanin.
    (Peptide available as ab16375)

  • General notes

    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.

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 or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Primary antibody notes

    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.
  • Clonality

    Polyclonal
  • Isotype

    IgG
  • Research areas

    • Cell Biology
    • Apoptosis
    • Nucleus
    • Lamins
    • Tags & Cell Markers
    • Subcellular Markers
    • Nucleus
    • Nuclear Envelope
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Intermediate Filaments
    • Class V
    • Lamins
    • Isotype/Loading Controls
    • Loading Controls
    • Lamin B1 - Nuclear
    • Cancer
    • Cell Death
    • Apoptosis
    • Nucleus
    • Lamins

Associated products

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Immunizing Peptide (Blocking)

    • Mouse Lamin B1 peptide (ab16375)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)

Applications

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.

Application Abreviews Notes
ICC/IF Use a concentration of 0.1 - 1 µg/ml.
WB Use a concentration of 0.1 µg/ml. Detects a band of approximately 68 kDa (predicted molecular weight: 66 kDa).

We recommend Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773).

IHC-P Use a concentration of 0.1 - 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

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.
  • Involvement in disease

    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.
  • Sequence similarities

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

    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.
  • Cellular localization

    Nucleus inner membrane.
  • Target information above from: UniProt accession P20700 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 396223 Chicken
    • Entrez Gene: 4001 Human
    • Entrez Gene: 16906 Mouse
    • Entrez Gene: 100513342 Pig
    • Entrez Gene: 116685 Rat
    • Entrez Gene: 379745 Xenopus laevis
    • Entrez Gene: 195816 Zebrafish
    • Omim: 150340 Human
    • SwissProt: P14731 Chicken
    • SwissProt: P20700 Human
    • SwissProt: P14733 Mouse
    • SwissProt: P70615 Rat
    • Unigene: 89497 Human
    • Unigene: 4105 Mouse
    • Unigene: 11362 Rat
    see all
  • Alternative names

    • 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

Images

  • Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
    Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: empty lane
    Lane 3: KO HAP1 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 react with  LMNB1 (Lamin B1) in wild type HAP1 cells. No band was observed when LMNB1 (Lamin B1) knockout samples were used. Ab16048 LMNB1 (Lamin B1) 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) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
    Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)Image courtesy of Marilena Ciciarello and Patrizia Lavia, University 'La Sapienza' CNR, Italy

    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)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)

    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

  • Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
    Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)

    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 in wild-type HAP1 cells. No band was observed 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.

  • Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
    Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
    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
    All lanes : 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?

  • Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
    Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)

    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).

  • Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
    Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)This image is courtesy of an anonymous Abreview
    Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048) at 1/1000 dilution + Human 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

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)This image is courtesy of an anonymous 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

  • Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
    Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)Image courtesy of Marilena Ciciarello and Patrizia Lavia, University 'La Sapienza' CNR, Italy

    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)

     

Protocols

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

    • Datasheet
    • SDS
  • References (625)

    Publishing research using ab16048? Please let us know so that we can cite the reference in this datasheet.

    ab16048 has been referenced in 625 publications.

    • Godavarthy PS  et al. The vascular bone marrow niche influences outcome in chronic myeloid leukemia via the E-selectin - SCL/TAL1 - CD44 axis. Haematologica 105:136-147 (2020). PubMed: 31018977
    • Spadotto V  et al. PRMT1-mediated methylation of the microprocessor-associated proteins regulates microRNA biogenesis. Nucleic Acids Res 48:96-115 (2020). PubMed: 31777917
    • Xie S  et al. Andrographolide Protects Against Adverse Cardiac Remodeling After Myocardial Infarction through Enhancing Nrf2 Signaling Pathway. Int J Biol Sci 16:12-26 (2020). PubMed: 31892842
    • Lowe DJ  et al. Chronic irradiation of human cells reduces histone levels and deregulates gene expression. Sci Rep 10:2200 (2020). PubMed: 32042076
    • Wintner O  et al. A Unified Linear Viscoelastic Model of the Cell Nucleus Defines the Mechanical Contributions of Lamins and Chromatin. Adv Sci (Weinh) 7:1901222 (2020). PubMed: 32328409
    View all Publications for this product

    Customer reviews and Q&As

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    Western blot abreview for Anti-Lamin B1 antibody - Nuclear Envelope Marker

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    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (Human coronary artery endothelial cells)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    10 µg
    Treatment
    10Gy of X-ray, collected 7 days after irradiation
    Specification
    Human coronary artery endothelial cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: rt°C
    Read More

    Sylwia Kabacik

    Verified customer

    Submitted Dec 17 2020

    Western blot abreview for Anti-Lamin B1 antibody - Nuclear Envelope Marker

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    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - nuclear (Gastric Cancer Cell Line)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    3 µg
    Specification
    Gastric Cancer Cell Line
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    Submitted Oct 29 2020

    Western blot abreview for Anti-Lamin B1 antibody - Nuclear Envelope Marker

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    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - nuclear (RAW264.7)
    Gel Running Conditions
    Reduced Denaturing
    Loading amount
    3 µg
    Specification
    RAW264.7
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
    Read More

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    Verified customer

    Submitted Oct 29 2020

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Lamin B1 antibody - Nuclear Envelope Marker

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    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (primary adult microvascular endothelial cells)
    Permeabilization
    Yes - 0.1% triton X
    Specification
    primary adult microvascular endothelial cells
    Blocking step
    Serum as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: RT°C
    Fixative
    Paraformaldehyde
    Read More

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    Submitted Sep 10 2020

    Western blot abreview for Anti-Lamin B1 antibody - Nuclear Envelope Marker

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    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Rat Tissue lysate - nuclear (soleus muscle)
    Gel Running Conditions
    Reduced Non-Denaturing (Native) (10)
    Loading amount
    10 µg
    Specification
    soleus muscle
    Blocking step
    No blocking step used for 12 hour(s) and 0 minute(s) · Concentration: 1/1000µg/mL · Temperature: 4°C
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    Inna Paramonova

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    Submitted Sep 26 2019

    Western blot abreview for Anti-Lamin B1 antibody - Nuclear Envelope Marker

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    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (melanoma)
    Gel Running Conditions
    Reduced Denaturing (10%)
    Loading amount
    10 µg
    Specification
    melanoma
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

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    Verified customer

    Submitted Jun 26 2019

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Lamin B1 antibody - Nuclear Envelope Marker

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    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (Human Melanoma)
    Permeabilization
    Yes - 0.2% Triton X100
    Specification
    Human Melanoma
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
    Fixative
    Paraformaldehyde
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    Verified customer

    Submitted Apr 30 2019

    Western blot abreview for Anti-Lamin B1 antibody - Nuclear Envelope Marker

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    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Mouse Cell lysate - nuclear (MEF)
    Gel Running Conditions
    Reduced Denaturing (12)
    Loading amount
    10 µg
    Specification
    MEF
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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    Submitted Jan 09 2019

    Western blot abreview for Anti-Lamin B1 antibody - Nuclear Envelope Marker

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    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Tissue lysate - nuclear (B Cells)
    Gel Running Conditions
    Reduced Denaturing (4-20%)
    Loading amount
    30 µg
    Specification
    B Cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

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    Verified customer

    Submitted Dec 18 2018

    Immunocytochemistry/ Immunofluorescence abreview for Anti-Lamin B1 antibody - Nuclear Envelope Marker

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    Abreviews
    Abreviews
    abreview image
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (HeLa Cells)
    Permeabilization
    Yes - Triton 0.3%
    Specification
    HeLa Cells
    Blocking step
    BSA as blocking agent for 15 minute(s) · Concentration: 1% · Temperature: 21°C
    Fixative
    Paraformaldehyde
    Read More

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