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
Overview
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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 asab16375) -
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
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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...
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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
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Related Products
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 |
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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). |
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IHC-P | Use a concentration of 0.1 - 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
Target
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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
modificationsB-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. - Information by UniProt
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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
see all -
Alternative names
- ADLD antibody
- lamin B1 antibody
- Lamin-B1 antibody
see all
Images
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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.
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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)
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
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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. -
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 withMouse 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)
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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Western blot - Anti-Lamin B1 antibody - Nuclear Envelope Marker (ab16048)This image is courtesy of an anonymous AbreviewAnti-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
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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.
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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
References (625)
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