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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (ab222391)

Submit a review Submit a question References (10)
Western blot - Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (ab222391)
  • Immunocytochemistry/ Immunofluorescence - Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (ab222391)
  • Flow Cytometry (Intracellular) - Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (ab222391)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (ab222391)
  • Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (ab222391)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [E398L] to Lamin B Receptor/LBR - BSA and Azide free
  • Suitable for: Flow Cyt (Intra), IHC-P, ICC/IF, WB
  • Knockout validated
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 647 APC PE Unconjugated

Overview

  • Product name

    Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free
    See all Lamin B Receptor/LBR primary antibodies

  • Description

    Rabbit monoclonal [E398L] to Lamin B Receptor/LBR - BSA and Azide free

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt (Intra), IHC-P, ICC/IF, WBmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Rat

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • IF: HeLa cells. WB: Jurkat cell lysate. HeLa: HEK-293 cell lysate. Flow Cyt (intra): HeLa cells.

  • General notes

    ab222391 is the carrier-free version of ab32535.

    Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

    This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

    Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.

    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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Mouse: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab222391 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
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 67 kDa (predicted molecular weight: 71 kDa).
Notes
Flow Cyt (Intra)
Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF
Use at an assay dependent concentration.
WB
Use at an assay dependent concentration. Detects a band of approximately 67 kDa (predicted molecular weight: 71 kDa).

Target

  • Function

    Anchors the lamina and the heterochromatin to the inner nuclear membrane.

  • Involvement in disease

    Defects 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 similarities

    Belongs to the ERG4/ERG24 family.

  • Post-translational
    modifications

    Phosphorylated 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 localization

    Nucleus inner membrane.
  • Information by UniProt

  • Database links

    • Alternative names

      • DHCR 14B antibody
      • DHCR14B antibody
      • Integral nuclear envelope inner membrane protein antibody
      • Lamin-B receptor antibody
      • LBR antibody
      • LBR_HUMAN antibody
      • LMN 2R antibody
      • LMN2R antibody
      • MGC9041 antibody
      • PHA antibody
      • PRO0650 antibody
      see all

    Images

    • Western blot - Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (ab222391)
      Western blot - Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (ab222391)
      All lanes : Anti-Lamin B Receptor/LBR antibody [E398L] (ab32535) at 1/500 dilution

      Lane 1 : Wild-type HEK-293 cell lysate
      Lane 2 : Human LBR (Lamin B Receptor) knockout HEK-293T cell lysate (ab257503)

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 71 kDa
      Observed band size: 58 kDa why is the actual band size different from the predicted?



      This data was developed using the same antibody clone in a different buffer formulation (ab32535).

      Lanes 1 - 2: Merged signal (red and green). Green - ab32535 observed at 58 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

      ab32535 was shown to react with Lamin B Receptor/LBR in wild-type HEK-293 cells in western blot. Loss of signal was observed when LBR knockout cell lysate ab257503 was used. Wild-type and LBR knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk before incubation with ab32535 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 500 dilution 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.

    • Immunocytochemistry/ Immunofluorescence - Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (ab222391)
      Immunocytochemistry/ Immunofluorescence - Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (ab222391)

      Ab32535, at a 1/500 dilution, staining Lamin B Receptor/LBR in HeLa cells by Immunofluorescence.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32535).

    • Flow Cytometry (Intracellular) - Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (ab222391)
      Flow Cytometry (Intracellular) - Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (ab222391)

      Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Lamin B Receptor/LBR with unpurified ab32535 at 1/20 dilutionc (10ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. 

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32535). 

       

       

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (ab222391)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (ab222391)
      IHC image of ab32535 staining in human breast cancer formalin fixed paraffin embedded tissue section, 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 20 mins. The section was then incubated with ab32535, 5µg/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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab32535).

    • Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (ab222391)
      Anti-Lamin B Receptor/LBR antibody [E398L] - BSA and Azide free (ab222391)

    References (10)

    Publishing research using ab222391? Please let us know so that we can cite the reference in this datasheet.

    ab222391 has been referenced in 10 publications.

    • Cundell MJ  et al. A PP2A-B55 recognition signal controls substrate dephosphorylation kinetics during mitotic exit. J Cell Biol 214:539-54 (2016). PubMed: 27551054
    • Mimura Y  et al. ELYS regulates the localization of LBR by modulating its phosphorylation state. J Cell Sci 129:4200-4212 (2016). WB, ICC/IF ; Human . PubMed: 27802161
    • Wang Y  et al. p32 is a novel target for viral protein ICP34.5 of herpes simplex virus type 1 and facilitates viral nuclear egress. J Biol Chem 289:35795-805 (2014). PubMed: 25355318
    • Clever M  et al. The nucleoporin ELYS/Mel28 regulates nuclear envelope subdomain formation in HeLa cells. Nucleus 3:187-99 (2012). ICC/IF . PubMed: 22555603
    • Doehle BP  et al. Vpu-deficient HIV strains stimulate innate immune signaling responses in target cells. J Virol 86:8499-506 (2012). PubMed: 22647704
    • Tseng LC & Chen RH Temporal control of nuclear envelope assembly by phosphorylation of lamin B receptor. Mol Biol Cell 22:3306-17 (2011). WB . PubMed: 21795390
    • Ahlenstiel CL  et al. Direct evidence of nuclear Argonaute distribution during transcriptional silencing links the actin cytoskeleton to nuclear RNAi machinery in human cells. Nucleic Acids Res : (2011). PubMed: 22064859
    • Gaudy-Marqueste C  et al. LBR mutation and nuclear envelope defects in a patient affected with Reynolds syndrome. J Med Genet 47:361-70 (2010). PubMed: 20522425
    • Maksimova N  et al. Neuroblastoma amplified sequence gene is associated with a novel short stature syndrome characterised by optic nerve atrophy and Pelger-Huet anomaly. J Med Genet : (2010). ICC/IF ; Human . PubMed: 20577004
    • Anderson DJ  et al. Recruitment of functionally distinct membrane proteins to chromatin mediates nuclear envelope formation in vivo. J Cell Biol 186:183-91 (2009). WB ; Human . PubMed: 19620630

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