PRODUCT CODE: ET1601-3

Filamin A Recombinant Rabbit Monoclonal Antibody [SA30-08] (ET1601-3)

  • Recombinant

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

Western blot analysis of Filamin A on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1601-3, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: MCF-7 cell lysate<br />
Lane 2: Jurkat cell lysate<br />
Lane 3: Hela cell lysate<br />
Lane 4: NIH/3T3 cell lysate
  • Western blot analysis of Filamin A on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1601-3, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.<br />
Positive control: <br />
Lane 1: MCF-7 cell lysate<br />
Lane 2: Jurkat cell lysate<br />
Lane 3: Hela cell lysate<br />
Lane 4: NIH/3T3 cell lysate
  • ICC staining of Filamin A in Hela cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1601-3, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Filamin A in AGS cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1601-3, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • ICC staining of Filamin A in HUVEC cells (green). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (ET1601-3, 1/50) for 1 hour at room temperature, washed with PBS. Alexa Fluor®488 Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. The nuclear counter stain is DAPI (blue).
  • Immunohistochemical analysis of paraffin-embedded human uterus tissue using anti-Filamin A antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-3, 1/100) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Immunohistochemical analysis of paraffin-embedded mouse uterus tissue using anti-Filamin A antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (ET1601-3, 1/400) for 30 minutes at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
  • Flow cytometric analysis of Filamin A was done on Hela cells. The cells were fixed, permeabilized and stained with the primary antibody (ET1601-3, 1/50) (blue). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Rabbit IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; red).
Western blot analysis of Filamin A on different lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (ET1601-3, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:5,000 dilution was used for 1 hour at room temperature.
Positive control:
Lane 1: MCF-7 cell lysate
Lane 2: Jurkat cell lysate
Lane 3: Hela cell lysate
Lane 4: NIH/3T3 cell lysate

Applications

  • WB

  • ICC

  • IF

  • IHC-P

  • FC

REACTIVITY

  • Human

  • Mouse

  • Rat

SPECIFICATIONS

Product Type

Recombinant Rabbit monoclonal primary

Product Name

Filamin A Recombinant Rabbit Monoclonal Antibody [SA30-08] (ET1601-3)

Immunogen

Synthetic peptide within c-terminal human filamin a.

Host

Rabbit

Positive Control

MCF-7 cell lysate, Jurkat cell lysate, Hela cell lysate, NIH/3T3 cell lysate, Hela, AGS, HUVEC, human uterus tissue, mouse uterus tissue.

Conjugation

Unconjugated

Clonality

Monoclonal

Clone Number

SA30-08

PROPERTIES

Form

Liquid

Storage Condition

Store at +4C after thawing. Aliquot store at -20C or -80C. Avoid repeated freeze / thaw cycles.

Storage Buffer

1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.

Concentration

1 ug/ul

PURIFICATION

Protein A affinity purified.

MOLECULAR WEIGHT

281 kDa

Isotype

IgG

APPLICATION DILUTION

  • WB

  • 1:500-1:5,000

  • ICC/IF

  • 1:50-1:200

  • IHC-P

  • 1:50-1:500

  • FC

  • 1:50-1:100

TARGET

UNIPROT #

PROTEIN NAME

Filamin A

SYNONYMS

ABP 280 antibody; ABP-280 antibody; Actin-binding protein 280 antibody; Alpha filamin antibody; Alpha-filamin antibody; APBX antibody; CSBS antibody; CVD1 antibody; Endothelial actin binding protein antibody; Endothelial actin-binding protein antibody; Filamin 1 antibody; Filamin A alpha antibody; Filamin A antibody; Filamin-1 antibody; Filamin-A antibody; FLN antibody; FLN-A antibody; FLN1 antibody; FLNA antibody; FLNA_HUMAN antibody; FMD antibody; MNS antibody; NHBP antibody; Non muscle filamin antibody; Non-muscle filamin antibody; OPD antibody; OPD1 antibody; OPD2 antibody; XLVD antibody; XMVD antibody

SEQUENCE SIMILARITIES

Belongs to the filamin family.

TISSUE SPECIFICITY

Ubiquitous.

POST-TRANSLATIONAL MODIFICATION

Phosphorylation at Ser-2152 is negatively regulated by the autoinhibited conformation of filamin repeats 19-21. Ligand binding induces a conformational switch triggering phosphorylation at Ser-2152 by PKA.; Phosphorylation extent changes in response to cell activation.; Polyubiquitination in the CH1 domain by a SCF-like complex containing ASB2 leads to proteasomal degradation. Prior dissociation from actin may be required to expose the target lysines. Ubiquitinated in endothelial cells by RNF213 downstream of the non-canonical Wnt signaling pathway, leading to its degradation by the proteasome.

SUBCELLULAR LOCATION

Cytoplasm.

FUNCTION

Caldesmon, Filamin 1, Nebulin and Villin are differentially expressed and regulated Actin binding proteins. Both muscular (CDh) and non-muscular (CDl) forms of Caldesmon have been identified and each has been shown to bind to Actin as well as to calmodulin and Myosin. CDh is expressed predominantly on thin filaments in smooth muscle, whereas CDl is widely expressed in non-muscle tissues and cells. Filamin 1, which is ubiquitously expressed and exists as a homodimer, functions to crosslink Actin to filaments. Nebulin is a large filamentous protein specific to muscle tissue that may function as a ruler for filament length. Several isoforms of Nebulin are produced by alternative exon usage. Villin is Ca2+-regulated and is the major structural component of the brush border of absorptive cells.