Overview

  • Product name

  • Description

    Rabbit polyclonal to Filamin B
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • Immunogen

    Recombinant fragment corresponding to Human Filamin B aa 30-239.
    Database link: O75369

  • Positive control

    • WB: 293T, A431, H1299, HeLa and HepG2 whole cell lysate (ab7900) IF: HeLa cells

Properties

Applications

Our Abpromise guarantee covers the use of ab97457 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
WB 1/500 - 1/3000. Predicted molecular weight: 278 kDa.
ICC/IF 1/100 - 1/200.

Target

  • Function

    Connects cell membrane constituents to the actin cytoskeleton. May promote orthogonal branching of actin filaments and links actin filaments to membrane glycoproteins. Anchors various transmembrane proteins to the actin cytoskeleton. Interaction with FLNA may allow neuroblast migration from the ventricular zone into the cortical plate. Various interactions and localizations of isoforms affect myotube morphology and myogenesis. Isoform 6 accelerates muscle differentiation in vitro.
  • Tissue specificity

    Ubiquitous. Isoform 1 and isoform 2 are expressed in placenta, bone marrow, brain, umbilical vein endothelial cells (HUVEC), retina and skeletal muscle. Isoform 1 is predominantly expressed in prostate, uterus, liver, thyroid, stomach, lymph node, small intestine, spleen, skeletal muscle, kidney, placenta, pancreas, heart, lung, platelets, endothelial cells, megakaryocytic and erythroleukemic cell lines. Isoform 2 is predominantly expressed in spinal cord, platelet and Daudi cells. Also expressed in thyroid adenoma, neurofibrillary tangles (NFT), senile plaques in the hippocampus and cerebral cortex in Alzheimer disease (AD). Isoform 3 and isoform 6 are expressed predominantly in lung, heart, skeletal muscle, testis, spleen, thymus and leukocytes. Isoform 4 and isoform 5 are expressed in heart.
  • Involvement in disease

    Note=Interaction with FLNA may compensate for dysfunctional FLNA homodimer in the periventricular nodular heterotopia (PVNH) disorder.
    Defects in FLNB are the cause of atelosteogenesis type 1 (AO1) [MIM:108720]; also known as giant cell chondrodysplasia or spondylohumerofemoral hypoplasia. Atelosteogenesis are lethal short-limb skeletal dysplasias with vertebral abnormalities, disharmonious skeletal maturation, poorly modeled long bones and joint dislocations.
    Defects in FLNB are the cause of atelosteogenesis type 3 (AO3) [MIM:108721]. Atelosteogenesis are short-limb lethal skeletal dysplasias with vertebral abnormalities, disharmonious skeletal maturation, poorly modeled long bones and joint dislocations. In AO3 recurrent respiratory insufficiency and/or infections usually result in early death.
    Defects in FLNB are the cause of boomerang dysplasia (BOOMD) [MIM:112310]. This is a perinatal lethal osteochondrodysplasia characterized by absence or underossification of the limb bones and vertebre. Boomerang dysplasia is distinguished from atelosteogenesis on the basis of a more severe defect in mineralisation, with complete absence of ossification in some limb elements and vertebral segments.
    Defects in FLNB are the cause of Larsen syndrome (LRS) [MIM:150250]. An osteochondrodysplasia characterized by large-joint dislocations and characteristic craniofacial abnormalities. The cardinal features of the condition are dislocations of the hip, knee and elbow joints, with equinovarus or equinovalgus foot deformities. Spatula-shaped fingers, most marked in the thumb, are also present. Craniofacial anomalies include hypertelorism, prominence of the forehead, a depressed nasal bridge, and a flattened midface. Cleft palate and short stature are often associated features. Spinal anomalies include scoliosis and cervical kyphosis. Hearing loss is a well-recognized complication.
    Defects in FLNB are the cause of spondylocarpotarsal synostosis syndrome (SCT) [MIM:272460]; also known as spondylocarpotarsal syndrome (SCT) or congenital synspondylism or vertebral fusion with carpal coalition or congenital scoliosis with unilateral unsegmented bar. The disorder is characterized by short stature and vertebral, carpal and tarsal fusions.
  • Sequence similarities

    Belongs to the filamin family.
    Contains 1 actin-binding domain.
    Contains 2 CH (calponin-homology) domains.
    Contains 24 filamin repeats.
  • Domain

    Comprised of a NH2-terminal actin-binding domain, 24 internally homologous repeats and two hinge regions. Repeat 24 and the second hinge domain are important for dimer formation. The first hinge region prevents binding to ITGA and ITGB subunits.
  • Post-translational
    modifications

    ISGylation prevents ability to interact with the upstream activators of the JNK cascade and inhibits IFNA-induced JNK signaling.
  • Cellular localization

    Cytoplasm > cytoskeleton. Polarized at the periphery of myotubes; Cytoplasm > cytoskeleton. Predominantly localized at actin stress fibers and Cytoplasm > cell cortex. Cytoplasm > cytoskeleton. Cytoplasm > myofibril > sarcomere > Z line. In differentiating myotubes, isoform 1, isoform 2 and isoform 3 are localized diffusely throughout the cytoplasm with regions of enrichment at the longitudinal actin stress fiber. In differentiated tubes, isoform 1 is also detected within the Z-lines.
  • Information by UniProt
  • Database links

  • Alternative names

    • ABP 278 antibody
    • ABP 280 homolog antibody
    • ABP-278 antibody
    • ABP-280 homolog antibody
    • ABP278 antibody
    • Actin binding like protein antibody
    • Actin binding protein 278 antibody
    • Actin-binding-like protein antibody
    • AOI antibody
    • Beta filamin antibody
    • Beta-filamin antibody
    • DKFZp686A1668 antibody
    • DKFZp686O033 antibody
    • Fh1 antibody
    • Filamin 1 actin binding protein 280 like antibody
    • Filamin 3 antibody
    • Filamin B antibody
    • Filamin B beta actin binding protein 278 antibody
    • Filamin homolog 1 antibody
    • Filamin-3 antibody
    • Filamin-B antibody
    • FLN B antibody
    • FLN-B antibody
    • FLN1L antibody
    • FLN3 antibody
    • FLNB antibody
    • FLNB_HUMAN antibody
    • LRS1 antibody
    • SCT antibody
    • TABP antibody
    • TAP antibody
    • Thyroid autoantigen antibody
    • Truncated ABP antibody
    • Truncated actin binding protein antibody
    • Truncated actin-binding protein antibody
    see all

Images

  • Anti-Filamin B antibody (ab97457) at 1/1000 dilution + A431 whole cell lysate at 30 µg

    Predicted band size: 278 kDa



    5% SDS PAGE.

    Secondary antibody - goat anti-rabbit HRP (ab6721)

  • Anti-Filamin B antibody (ab97457) at 1/1000 dilution + NIH3T3 whole cell lysate at 30 µg

    Predicted band size: 278 kDa



    5% SDS PAGE.

     

    Secondary antibody - goat anti-rabbit HRP (ab6721)

  • ab97547, at a 1/200 dilution, staining Filamin B in paraformaldehyde-fixed HeLa cells by Immunofluorescence analysis. Right image is merged with a DNA probe.

  • Immunofluorescence analysis of Human trabecular meshwork cells, staining Filamin B with ab97457.

    Left panel: Filamin B staining alone. Right panel: Filamin B staining (red) merged with F-actin staining (green).

    Cells were treated with dexamethasone, before fixing in paraformaldehyde and permeabilizing wih 0.2% Triton X-100. Cells were incubated with primary antibody (10 µg/ml) and AlexaFluor®488-conjugated phalloidin. Filamin B staining was detected using an AlexaFluor®546-conjugated goat anti-rabbit IgG.

References

This product has been referenced in:

  • Tang B  et al. PEA15 promotes liver metastasis of colorectal cancer by upregulating the ERK/MAPK signaling pathway. Oncol Rep 41:43-56 (2019). Read more (PubMed: 30365128) »
  • Clark R  et al. Comparative genomic and proteomic analysis of cytoskeletal changes in dexamethasone-treated trabecular meshwork cells. Mol Cell Proteomics 12:194-206 (2013). ICC/IF . Read more (PubMed: 23105009) »
See all 2 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (fibroblast)
Permeabilization
Yes - 0.3% Triton X-100 in blocking buffer
Specification
fibroblast
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Mar 15 2018

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (lymphocytes)
Total protein in input
35 µg
Immuno-precipitation step
Protein A
Specification
lymphocytes

Abcam user community

Verified customer

Submitted Feb 25 2014

Application
Western blot
Sample
Human Cell lysate - whole cell (lymphocytes)
Gel Running Conditions
Reduced Denaturing (4-12% Bis-Tris)
Loading amount
35 µg
Specification
lymphocytes
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 30 2014

Answer

Thank you for submitting the Abreview of ab97457. Your review has now been published on our website. Since unexpected results were obtained with this antibody in a tested species and application, we would like to follow up to see if the results can be improved. Since the protein target has such a high molecular weight (predicted around 278 kDa), it may be necessary to alter the protocol to ensure that the protein transfers to the membrane. You can add 0.1% SDS to the transfer buffer and decrease the methanol percentage to 10% or less to improve transfer. Decreasing the antibody dilution to 1:500 and loading 20-30 ug of protein will ensure that the 278 kDa protein can be detected after transfer. You mentioned that the smaller bands may be cleavage fragments- what kind of protease inhibitors were used? We do guarantee this antibody to work in Western blot with human samples according to our Abpromise, so if these suggestions do not improve the results we would be happy to send a replacement or issue a credit or refund. I look forward to hearing from you. Please let me know if you have any questions or if there is anything else that we can do for you.

Read More
Abreviews
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293,HELA and JURKAT)
Loading amount
10 µg
Specification
HEK293,HELA and JURKAT
Gel Running Conditions
Reduced Denaturing (4-20%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Sep 07 2011

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