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Western blot
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Immunocytochemistry/ Immunofluorescence - Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
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Immunocytochemistry - Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
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Product Name
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Lamin B1 antibody - Nuclear Envelope Marker
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See all Lamin B1 - Nuclear Envelope Marker antibodies (7)...
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Product type
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Primary antibodies
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Description
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Rabbit polyclonal to Lamin B1 - Nuclear Envelope Marker
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Immunogen
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Synthetic peptide conjugated to KLH derived from within residues 400 - 500 of Mouse Lamin B1.
(Peptide available as ab16375.)
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Reacts with
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Hu, Ms, Rat, InMtj
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Predicted to react
(species key)
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Chk, Xl, Zfsh
(Due to sequence homology)
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Specificity
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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.
N.B. In-house QC testing has shown that Batches 112793 and 112795 do not detect Lamin B1 in 3T3 cells by Western blot. Batches 112787, 112791 and 151601 do detect Lamin B1 in Mouse cell lysates. Batch 112793 does not appear to detect Lamin B1 in Rat, Xenopus or Zebrafish lysate and only weakly detects Lamin B1 in Chicken liver lysate.
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Tested applications
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ICC/IF, IHC-P, WB
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Abreviews
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Application notes
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Recommended dilutions
IHC-P: Use at a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF: Use at a concentration of 1 µg/ml.
WB: Use at a concentration of 0.1 µg/ml. Detects a band of approximately 68-70 kDa (predicted molecular weight: 66 kDa). Can be blocked with Lamin B1 peptide (ab16375).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
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Cellular localization
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Nuclear membrane
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Research areas
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Signal Transduction >> Cytoskeleton / ECM >> Cytoskeleton >> Intermediate Filaments >> Class V >> Lamins
Tags & Cell Markers >> Subcellular Markers >> Nucleus >> Nuclear Envelope
Cell Biology >> Apoptosis >> Nucleus >> Lamins
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Relevance
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An important part of the nucleus is formed by nuclear lamina. Nuclear lamins form a network of filaments at the nucleoplasmic site of the nuclear membrane. Two main subtypes of nuclear lamins can be distinguished, i.e. A type lamins and B type lamins. The A type lamins comprise a set of three proteins arising from the same gene by alternative splicing, i.e. lamin A, lamin C and lamin Adel10, while the B-type lamins include two proteins arising from two distinct genes, i.e. lamin B1 and lamin B2.
The nuclear lamins comprise a unique subclass of the intermediate filament protein family. They share a molecular domain organisation with the other intermediate filament proteins in that they are fibrous molecules that have an aminoterminal globular head, a central rod of alpha helices and a carboxy terminal globular domain.
Many biochemical and molecular features of lamins have been studied, but their functions remain still largely undetermined. One of the functions ascribed to the lamina is the maintenance of the structural integrity of the nucleus.
Besides interactions with the nuclear membrane and other intermediate filaments, lamins interact with the nuclear chromatin. Eukaryotic chromatin is organised into loops, which are attached to the nuclear matrix. This organisation is thought to contribute to compaction of the chromatin and regulation of gene expression. Lamins, as part of the nuclear matrix, may be involved in these processes since chromatin binding sites have been detected in both A and B type lamins.
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.
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Database links
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The links below go to external sites and will open in a new browser window |
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Raised in
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Rabbit
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Clonality
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Polyclonal
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Isotype
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IgG
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Purity
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Immunogen affinity purified
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Storage buffer
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Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4 Material safety datasheet (MSDS) for this product:
Sodium Azide MSDS
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Form
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Liquid
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Concentration
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Batch dependent within range: 0.20 - 0.50 mg/ml
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Storage instructions
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Store at +4°C short term (1-2 weeks). Aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this Lamin B1 antibody - Nuclear Envelope Marker is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for Lamin B1 antibody - Nuclear Envelope Marker images, references, products related to ab16048 and other tools.
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Lamin B1 antibody - Nuclear Envelope Marker images:
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 Western blot
All lanes : Lamin B1 antibody - Nuclear Envelope Marker (ab16048) at 1/1000 dilution
Lane 1 : Hela whole cell lysate
Lane 2 : Hela whole cell lysate with 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?)
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Immunocytochemistry/ Immunofluorescence - 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).
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Immunocytochemistry - Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
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) Image courtesy of Marilena Ciciarello and Patrizia Lavia, University 'La Sapienza' CNR, Italy
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Immunocytochemistry - Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
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) Image courtesy of Marilena Ciciarello and Patrizia Lavia, University 'La Sapienza' CNR, Italy
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Western blot - Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
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
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
IHC image of Lamin B1 staining in human liver FFPE section, performed on a BondTM 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 8 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.
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References for Lamin B1 antibody - Nuclear Envelope Marker (ab16048)
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This product has been used in: (two most recent references)
Chevanne M et al. Inhibition of PARP activity by PJ-34 leads to growth impairment and cell death associated with aberrant mitotic pattern and nucleolar actin accumulation in M14 melanoma cell line. J Cell Physiol 222:401-10 (2010). WB; Human. PubMed: 19890834
Roth RJ et al. MAPK phosphatase-1 facilitates the loss of oxidative myofibers associated with obesity in mice. J Clin Invest 119:3817-29 (2009). WB; Mouse. PubMed: 19920356
See all 31 publication references for this product.
If you publish research using ab16048 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to Lamin B1
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