Recombinant Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab133741)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR8985(B)] to Lamin B1 - Nuclear Envelope Marker
- Suitable for: IP, ICC/IF, WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker
See all Lamin B1 primary antibodies -
Description
Rabbit monoclonal [EPR8985(B)] to Lamin B1 - Nuclear Envelope Marker -
Host species
Rabbit -
Tested applications
Suitable for: IP, ICC/IF, WB, IHC-Pmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. within Human Lamin B1 aa 500 to the C-terminus. The exact sequence is proprietary.
Database link: P20700 -
Positive control
- WB: Hap1, HeLa, Jurkat, Molt4, Y79, Caco 2, C6, Raw264.7, PC-12 and NIH/3T3 cell lysates. Mouse brain,heart, kidney and spleen; and Rat brain, heart and spleen lysates. IHC-P: Human colon, liver and transitional cell carcinoma of the bladder tissues. ICC/IF: Ramos cells, HAP1-LMNB1 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
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 long term. Avoid freeze / thaw cycle. -
Dissociation constant (KD)
KD = 1.95 x 10 -10 M Learn more about KD -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR8985(B) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
- Alexa Fluor® 488 Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab194106)
- Alexa Fluor® 647 Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab194108)
- HRP Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Loading Control (ab194109)
- Alexa Fluor® 594 Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab216723)
- Anti-Lamin B1 antibody [EPR8985(B)] - BSA and Azide free (ab220797)
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Compatible Secondaries
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab133741 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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IP |
1/20.
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ICC/IF | (4) |
Use a concentration of 1 µg/ml.
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WB | (9) |
1/1000 - 1/10000. Detects a band of approximately 70 kDa (predicted molecular weight: 66 kDa).
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IHC-P | (1) |
1/300. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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Notes |
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IP
1/20. |
ICC/IF
Use a concentration of 1 µg/ml. |
WB
1/1000 - 1/10000. Detects a band of approximately 70 kDa (predicted molecular weight: 66 kDa). |
IHC-P
1/300. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 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: 4001 Human
- Entrez Gene: 16906 Mouse
- Entrez Gene: 116685 Rat
- Omim: 150340 Human
- SwissProt: P20700 Human
- SwissProt: P14733 Mouse
- SwissProt: P70615 Rat
- Unigene: 89497 Human
see all -
Alternative names
- ADLD antibody
- lamin B1 antibody
- Lamin-B1 antibody
see all
Images
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All lanes : Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab133741) at 1/1000 dilution
Lane 1 : Wild-type HeLa cell lysate
Lane 2 : LMNB1 knockout HeLa cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 66 kDa
Observed band size: 66-70 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab133741 observed at 66-70 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab133741 was shown to react with LMNB1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab255404 (knockout cell lysate ab263825) was used. Wild-type HeLa and LMNB1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab133741 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 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 in 20000 dilution for 1 hour at room temperature before imaging.
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Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab133741)
Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Ramos (Human Burkitt's lymphoma cell line) cells labeling Lamin B1 with ab133741 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/200 dilution (green). Nuclear envelope staining on Ramos cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).
The negative controls are as follows:
-ve control 1: ab133741 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2: ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/200 dilution. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab133741)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human transitional cell carcinoma of the bladder tissue labeling Lamin B1 with purified ab133741 at 1/300. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. A prediluted HRP-polymer conjugated anti-rabbit IgG was used as the secondary antibody. Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunocytochemistry/ Immunofluorescence - Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab133741)
ab133741 staining Lamin B1 in wild-type HAP1 cells (top panel) and Lamin B1 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 ab133741 at 1μg/ml dilution and ab195889 at 1/250 dilution (shown in pseudo-color red) overnight at +4°C. The cells were then incubated with ab150081 (Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488)) at 1/1000 dilution for 1 hour. Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Lanes 1 & 3 : Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab133741) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate at 20 µg
Lanes 2 & 4 : Empty
Lane 3 : LMNB1 knockout HAP1 whole cell lysate at 20 µg
Predicted band size: 66 kDaLanes 1 - 4: Merged signal (red and green). Green - ab133741 observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab133741 was shown to specifically react with Lamin B1 in wild type HAP1 cells. No band was observed when knockout samples were used. Wild-type and Lamin B1 knockout samples were subjected to SDS-PAGE. Ab133741 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.
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Lanes 1 & 3 : Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab133741)
Lane 1 : Wild-type HAP1 whole cell lysate at 20 µg
Lane 2 : Empty
Lane 3 : Lamin B1 knockout HAP1 cell lysate at 20 µg
Predicted band size: 66 kDaLanes 1 - 3: Merged signal (red and green).
Green - Target observed at 70 kDa. Red - loading control, ab8245, observed at 37 kDa.
This western blot image is a comparison between ab133741 and a competitor's discontinued goat polyclonal antibody.
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Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab133741) at 1/1000 dilution + GST-tagged Recombinant Human Lamin B1 protein (aa 1 to 586) at 0.015 µg with 5% NFDM/TBST
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 66 kDa
Observed band size: 100 kDa why is the actual band size different from the predicted?
Exposure time: 1 secondRecombinant Human Lamin B1 protein (ab114163)
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All lanes : Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab133741) at 1/50000 dilution (purified)
Lane 1 : Jurkat (Human T cell leukemia cells from peripheral blood)
cell lysate
Lane 2 : Molt-4 (Human lymphoblastic leukemia cell line) cell lysate
Lane 3 : Y79 (Human retinoblastoma cell line) cell lysate
Lane 4 : Caco-2 (Human colorectal adenocarcinoma cells) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 66 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab133741)
Immunohistochemical staining of paraffin embedded Mouse Cerebral cortex with purified ab133741 at a working dilution of 1/300. The secondary antibody used is a HRP polymer for rabbit IgG. Nuclear envelope staining on neuron cells of Cerebral cortex tissue is observed. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
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All lanes : Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab133741) at 1/50000 dilution (purified)
Lane 1 : C6 (Rat glial tumor cells) cell lysate
Lane 2 : PC12 (Rat adrenal gland pheochromocytoma) cell lysate
Lane 3 : NIH/3T3 (Mouse embyro fibroblast cells) cell lysate
Lane 4 : RAW264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 66 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?Blocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab133741) at 1/10000 dilution (purified) + Jurkat (Human T cell leukemia cells from peripheral blood) cell lysate at 10 µg
Secondary
Peroxidase conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution
Predicted band size: 66 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab133741)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human liver tissue labelling Lamin B1 with unpurified ab133741 at 1/250.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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All lanes : Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab133741) at 1/10000 dilution
Lane 1 : Mouse brain lysates
Lane 2 : Mouse heart lysates
Lane 3 : Mouse kidney lysates
Lane 4 : Mouse spleen lysates
Lane 5 : Rat brain lysates
Lane 6 : Rat heart lysates
Lane 7 : Rat spleen lysates
Lane 8 : C6 (Rat glial tumor cells) whole cell lysates
Lane 9 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates
Lane 10 : NIH/3T3 (Mouse embyro fibroblast cells)
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution
Predicted band size: 66 kDa
Observed band size: 70 kDa why is the actual band size different from the predicted?Blocking and Diluting buffer and concentration: 5% NFDM/TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B1 antibody [EPR8985(B)] - Nuclear Envelope Marker (ab133741)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue labelling Lamin B1 with unpurified ab133741 at 1/250.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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ab133741 (purified) at 1/20 immunoprecipitating Lamin B1 in Jurkat cells (Lane 1). For western blotting, ab133741 was used at 1/1000 dilution and an HRP-conjugated goat anti-rabbit IgG was used as the secondary antibody (1/1500).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (126)
ab133741 has been referenced in 126 publications.
- Yang L et al. Effect of the isoflavone corylin from cullen corylifolium on colorectal cancer growth, by targeting the STAT3 signaling pathway. Phytomedicine 80:153366 (2021). PubMed: 33080566
- Kim S et al. Characterization of ferroptosis in kidney tubular cell death under diabetic conditions. Cell Death Dis 12:160 (2021). PubMed: 33558472
- Chen D et al. Downregulation of long non-coding RNA MR4435-2HG suppresses breast cancer progression via the Wnt/ß-catenin signaling pathway. Oncol Lett 21:373 (2021). PubMed: 33777197
- Kang JW et al. PUMA facilitates EMI1-promoted cytoplasmic Rad51 ubiquitination and inhibits DNA repair in stem and progenitor cells. Signal Transduct Target Ther 6:129 (2021). PubMed: 33785736
- Todkar K et al. Selective packaging of mitochondrial proteins into extracellular vesicles prevents the release of mitochondrial DAMPs. Nat Commun 12:1971 (2021). PubMed: 33785738