Key features and details
- Mouse monoclonal [BBmLBR 12.F8] to Lamin B Receptor/LBR
- Suitable for: ICC/IF, WB, IHC-P
- Knockout validated
- Reacts with: Mouse, Human
- Isotype: IgG2b
Product nameAnti-Lamin B Receptor/LBR antibody [BBmLBR 12.F8]
See all Lamin B Receptor/LBR primary antibodies
DescriptionMouse monoclonal [BBmLBR 12.F8] to Lamin B Receptor/LBR
Tested applicationsSuitable for: ICC/IF, WB, IHC-Pmore details
Species reactivityReacts with: Mouse, Human
Recombinant fragment within Mouse Lamin B Receptor/LBR aa 1-250 (N terminal). The exact sequence is proprietary.
- ICC/IF: HEK-293 cells. WB: MEF-1 whole cell lysate IHC-P: Human Duodenum
This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact firstname.lastname@example.org.
Storage instructionsShipped 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 bufferPreservative: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine
Concentration information loading...
PurityProtein G purified
Clone numberBBmLBR 12.F8
Light chain typekappa
KO cell lysates
Our Abpromise guarantee covers the use of ab232731 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 10 µg/ml.|
|WB||Use a concentration of 1 µg/ml. Detects a band of approximately 65 kDa (predicted molecular weight: 71 kDa).|
|IHC-P||Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.|
FunctionAnchors the lamina and the heterochromatin to the inner nuclear membrane.
Involvement in diseaseDefects in LBR are a cause of Pelger-Huet anomaly (PHA) [MIM:169400]. PHA is an autosomal dominant inherited abnormality of neutrophils, characterized by reduced nuclear segmentation and an apparently looser chromatin structure. Heterozygotes show hypolobulated neutrophil nuclei with coarse chromatin. Presumed homozygous individuals have ovoid neutrophil nuclei, as well as varying degrees of developmental delay, epilepsy, and skeletal abnormalities.
Defects in LBR are the cause of hydrops-ectopic calcification-moth-eaten skeletal dysplasia (HEM) [MIM:215140]; also known as Greenberg skeletal dysplasia. HEM is a rare autosomal recessive chondrodystrophy characterized by early in utero lethality and, therefore, considered to be nonviable. Affected fetuses typically present with fetal hydrops, short-limbed dwarfism, and a marked disorganization of chondro-osseous calcification and may present with polydactyly and additional nonskeletal malformations.
Defects in LBR may be a cause of Reynolds syndrome (REYNS) [MIM:613471]. It is a syndrome specifically associating limited cutaneous systemic sclerosis and primary biliray cirrhosis. It is characterized by liver disease, telangiectasia, abrupt onset of digital paleness or cyanosis in response to cold exposure or stress (Raynaud phenomenon), and variable features of scleroderma. The liver disease is characterized by pruritis, jaundice, hepatomegaly, increased serum alkaline phosphatase and positive serum mitochondrial autoantibodies, all consistent with primary biliary cirrhosis.
Sequence similaritiesBelongs to the ERG4/ERG24 family.
modificationsPhosphorylated by CDK1 protein kinase in mitosis when the inner nuclear membrane breaks down into vesicles that dissociate from the lamina and the chromatin. It is phosphorylated by different protein kinases in interphase when the membrane is associated with these structures. Phosphorylation of LBR and HP1 proteins may be responsible for some of the alterations in chromatin organization and nuclear structure which occur at various times during the cell cycle.
Cellular localizationNucleus inner membrane.
- Information by UniProt
- DHCR 14B antibody
- DHCR14B antibody
- Integral nuclear envelope inner membrane protein antibody
All lanes : Anti-Lamin B Receptor/LBR antibody [BBmLBR 12.F8] (ab232731) at 1 µg/ml
Lane 1 : Wild-type HEK-293 whole cell lysate
Lane 2 :
Human LBR (Lamin B Receptor) knockout HEK293T cell lysate (ab257503)
Lane 3 : Wild-type MEF-1 whole cell lysate
Lane 4 : LBR knockout MEF-1 whole cell lysate
Lysates/proteins at 20 µg per lane.
All lanes : Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 71 kDa
Observed band size: 58 kDa why is the actual band size different from the predicted?
Lanes 1 - 4: Merged signal (red and green). Green - ab232731 observed at 58 kDa. Red - loading control, ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) observed at 37kDa.
ab232731 was shown to react with Lamin B Receptor (LBR) in wild-type HEK-293 and MEF-1 cells in western blot. Loss of signal was observed when LBR knockout samples were used. Wild-type and LBR knockout (ab257503) HEK-293 cell lysates and wild-type and LBR knockout MEF-1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk before incubation with ab232731 and ab181602 (Rabbit Anti-GAPDH antibody [EPR16891]) overnight at 4°C at 1 µg/ml and a 1 in 20000 dilution respectively. Blots were incubated 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 in 20000 dilution for 1 hour at room temperature before imaging.
LBR knockout MEF-1 samples were kindly provided by the Brian Burke laboratory, A-Star Institute, Singapore.
IHC image of Lamin B receptor staining in a section of formalin-fixed paraffin-embedded normal human duodenum* performed on a Leica 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 20mins. The section was then incubated with ab232731, 0.05µg/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. The inset secondary-only control image is taken from an identical assay without primary antibody.
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.
ab232731 staining Lamin B Receptor in HEK-293 cells. The cells were fixed with 4% paraformaldehyde (10min), permeabilized with 0.1%PBS-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 overnight at +4°C with ab232731 at 10µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labeled with DAPI (shown in blue).
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
All lanes : Anti-Histone H4 antibody [EPR16599] - BSA and Azide free (ab232371) at 1 µg/ml
Lane 1 : MEF-1 wild-type whole cell lysate
Lane 2 : MEF-1 LBR knockout whole cell lysate
Lysates/proteins at 10 µg per lane.
Performed under reducing conditions.
Predicted band size: 71 kDa
Observed band size: 65 kDa why is the actual band size different from the predicted?
This blot was produced using a 4-12% Bis-tris under the MOPS buffer system. The gel was run at 200V for 55 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was blocked for an hour using 3% milk before ab232731 and ab181602 (Rabbit anti-GAPDH loading control) were incubated overnight at 4°C at a 1ug/ml concentration and 1/20000 dilution respectively. Antibody binding was detected using Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (ab216772) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed (ab216777) secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
Cell samples kindly provided by the Brian Burke laboratory, A-Star Institute, Singapore.
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
ab232731 has not yet been referenced specifically in any publications.