Overview

  • Product name

    Anti-SNF5/SMARCB1 antibody [EPR20189]
    See all SNF5/SMARCB1 primary antibodies
  • Description

    Rabbit monoclonal [EPR20189] to SNF5/SMARCB1
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, Flow Cyt, IP, ICC/IF, WBmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human SNF5/SMARCB1 aa 350 to the C-terminus. The exact sequence is proprietary.
    Database link: Q12824

  • Positive control

    • WB: HeLa, Jurkat, PC-12 and NIH-3T3 whole cell lysates; Human fetal brain, fetal heart and fetal spleen lysates; Mouse and rat brain and spleen lysates. IHC-P: Human, mouse and rat kidney tissues. ICC/IF: HeLa cells. Flow Cyt: HeLa cells. IP: HeLa whole cell lysate.
  • General notes

     This product was previously labelled as SNF5

     

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab222519 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
IHC-P 1/500. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt 1/500.
IP 1/30.
ICC/IF 1/100.
WB 1/1000. Detects a band of approximately 44 kDa (predicted molecular weight: 44 kDa).

Target

  • Function

    Core component of the BAF (hSWI/SNF) complex. This ATP-dependent chromatin-remodeling complex plays important roles in cell proliferation and differentiation, in cellular antiviral activities and inhibition of tumor formation. The BAF complex is able to create a stable, altered form of chromatin that constrains fewer negative supercoils than normal. This change in supercoiling would be due to the conversion of up to one-half of the nucleosomes on polynucleosomal arrays into asymmetric structures, termed altosomes, each composed of 2 histones octamers. Stimulates in vitro the remodeling activity of SMARCA4/BRG1/BAF190A. Involved in activation of CSF1 promoter. Belongs to the neural progenitors-specific chromatin remodeling complex (npBAF complex) and the neuron-specific chromatin remodeling complex (nBAF complex). During neural development a switch from a stem/progenitor to a post-mitotic chromatin remodeling mechanism occurs as neurons exit the cell cycle and become committed to their adult state. The transition from proliferating neural stem/progenitor cells to post-mitotic neurons requires a switch in subunit composition of the npBAF and nBAF complexes. As neural progenitors exit mitosis and differentiate into neurons, npBAF complexes which contain ACTL6A/BAF53A and PHF10/BAF45A, are exchanged for homologous alternative ACTL6B/BAF53B and DPF1/BAF45B or DPF3/BAF45C subunits in neuron-specific complexes (nBAF). The npBAF complex is essential for the self-renewal/proliferative capacity of the multipotent neural stem cells. The nBAF complex along with CREST plays a role regulating the activity of genes essential for dendrite growth (By similarity). Plays a key role in cell-cycle control and causes cell cycle arrest in G0/G1. Also involved in vitamin D-coupled transcription regulation via its association with the WINAC complex, a chromatin-remodeling complex recruited by vitamin D receptor (VDR), which is required for the ligand-bound VDR-mediated transrepression of the CYP27B1 gene.
  • Involvement in disease

    Defects in SMARCB1 are a cause of rhabdoid tumor (RDT) [MIM:609322]; also known as malignant rhabdoid tumor (MRT). RDT are a highly malignant group of neoplasms that usually occur in early childhood. SMARCB1/INI1 is also frequently inactivated in epithelioid sarcomas.
    Defects in SMARCB1 are a cause of schwannomatosis (SCHWA) [MIM:162091]; also called congenital cutaneous neurilemmomatosis. Schwannomas are benign tumors of the peripheral nerve sheath that usually occur singly in otherwise normal individuals. Multiple schwannomas in the same individual suggest an underlying tumor-predisposition syndrome. The most common such syndrome is NF2. The hallmark of NF2 is the development of bilateral vestibular-nerve schwannomas; but two-thirds or more of all NF2-affected individuals develop schwannomas in other locations, and dermal schwannomas may precede vestibular tumors in NF2-affected children. There have been several reports of individuals with multiple schwannomas who do not show evidence of vestibular schwannoma. Clinical report suggests that schwannomatosis is a clinical entity distinct from other forms of neurofibromatosis.
  • Sequence similarities

    Belongs to the SNF5 family.
  • Post-translational
    modifications

    Phosphorylated upon DNA damage, probably by ATM or ATR.
  • Cellular localization

    Nucleus.
  • Information by UniProt
  • Database links

  • Alternative names

    • BAF47 antibody
    • BRG1-associated factor 47 antibody
    • hSNF5 antibody
    • INI1 antibody
    • Integrase interactor 1 protein antibody
    • Malignant rhabdoid tumor suppressor antibody
    • RDT antibody
    • RTPS1 antibody
    • Sfh1p antibody
    • SMARCB1 antibody
    • SNF5 homolog antibody
    • SNF5_HUMAN antibody
    • SNF5L1 antibody
    • Snr1 antibody
    • Sucrose nonfermenting yeast homolog like 1 antibody
    • SWI/SNF complex component SNF5 antibody
    • SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily B member 1 antibody
    • SWI10 antibody
    • Transcription factor TYE4 antibody
    • Transcription regulatory protein SNF5 antibody
    • TYE4 antibody
    see all

Images

  • All lanes : Anti-SNF5/SMARCB1 antibody [EPR20189] (ab222519) at 1/1000 dilution

    Lane 1 : HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 2 : G-401 (Human rhabdoid tumor kidney epithelial cell line) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 44 kDa
    Observed band size: 44 kDa


    Exposure time: 10 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Negative control: G-401 (PMID: 19789351).

     

  • All lanes : Anti-SNF5/SMARCB1 antibody [EPR20189] (ab222519) at 1/1000 dilution

    Lane 1 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg
    Lane 2 : PC-12 (rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg
    Lane 3 : NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate at 10 µg
    Lane 4 : Human fetal brain lysate at 10 µg
    Lane 5 : Human fetal heart lysate at 10 µg
    Lane 6 : Human fetal spleen lysate at 10 µg
    Lane 7 : Mouse brain lysate at 10 µg
    Lane 8 : Mouse spleen lysate at 10 µg
    Lane 9 : Rat brain lysate at 10 µg
    Lane 10 : Rat spleen lysate at 10 µg

    Secondary
    Lanes 1-3 & 7-10 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
    Lanes 4-6 : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 44 kDa
    Observed band size: 44 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times : Lane 1-3: 2 seconds; Lanes 4-5/7: 15 seconds; Lane 6/8-9: 4 seconds; Lane 10: 1 minute.

  • SNF5/SMARCB1 was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab222519 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab222519 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

    Lane 1: HeLa whole cell lysate 10 μg (input).
    Lane 2: ab222519 IP in HeLa whole cell lysate,
    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab222519 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST
    Exposure time: 10 seconds.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling SNF5/SMARCB1 with ab222519 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), Ready to use. Nuclear staining in human kidney is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunohistochemical analysis of paraffin-embedded mouse kidney tissue labeling SNF5/SMARCB1 with ab222519 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), Ready to use. Nuclear staining in mouse kidney is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunohistochemical analysis of paraffin-embedded rat kidney tissue labeling SNF5/SMARCB1 with ab222519 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP), Ready to use. Nuclear staining in rat kidney is observed. Counter stained with hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells labeling SNF5/SMARCB1 with ab222519 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear staining in HeLa cell line. 

    The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) (red) at 1/200 dilution.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    Negative control: G-401 cell line (PMID:19789351).

  • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol-permeablized G-401 (Human rhabdoid tumor kidney epithelial cell line, Left) and HeLa (Human epithelial cell line from cervix adenocarcinoma, Right) cells labeling SNF5/SMARCB1 with ab222519 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/2000 dilution was used as the secondary antibody.

    Negative control: G-401 (PMID:19789351)

References

ab222519 has not yet been referenced specifically in any publications.

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