Filamin A Recombinant Rabbit Monoclonal Antibody [SA30-08]
cat.: ET1601-3
| Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
|---|---|
| Species reactivity: | Human, Mouse, Rat, Monkey |
| Applications: | WB, IF-Cell, IF-Tissue, IHC-P, FC |
| Clonality: | Monoclonal |
| Clone number: | SA30-08 |
| Form: | Liquid |
| Storage condition: | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage buffer: | 1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide. |
| Concentration: | 1ug/ul |
| Purification: | Protein A affinity purified. |
| Molecular weight: | Predicted band size: 281 kDa |
| Isotype: | IgG |
| Immunogen: | Synthetic peptide within Human Filamin A aa 2,598-2,647 / 2,647. |
| Positive control: | HeLa cell lysate, HEK-293 cell lysate, A549 cell lysate, NIH/3T3 cell lysate, C2C12 cell lysate, C6 cell lysate, COS-1 cell lysate, HeLa, C2C12, C6, human uterus tissue, mouse uterus tissue. |
| Subcellular location: | Cytoplasm. |
| Recommended Dilutions:
WB IF-Cell IF-Tissue IHC-P FC |
1:2,000 1:200-1:500 1:50 1:100-1:400 1:1,000 |
| Uniprot #: | SwissProt: P21333 Human | Q8BTM8 Mouse Gene ID: 293860 Rat |
| Alternative names: | ABP 280 ABP-280 Actin-binding protein 280 Alpha filamin Alpha-filamin APBX CSBS CVD1 Endothelial actin binding protein Endothelial actin-binding protein Filamin 1 Filamin A alpha Filamin A Filamin-1 Filamin-A FLN FLN-A FLN1 FLNA FLNA_HUMAN FMD MNS NHBP Non muscle filamin Non-muscle filamin OPD OPD1 OPD2 XLVD XMVD |
Images
|
Fig1:
Western blot analysis of Filamin A on different lysates with Rabbit anti-Filamin A antibody (ET1601-3) at 1/2,000 dilution. Lane 1: HeLa cell lysate Lane 2: HEK-293 cell lysate Lane 3: A549 cell lysate Lane 4: NIH/3T3 cell lysate Lane 5: C2C12 cell lysate Lane 6: C6 cell lysate Lane 7: COS-1 cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 281 kDa Observed band size: 281 kDa Exposure time: 3 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-3) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
|
Fig2:
Western blot analysis of Filamin A on different lysates with Rabbit anti-Filamin A antibody (ET1601-3) at 1/2,000 dilution. Lane 1: HAP1-parental cell lysate Lane 2: HAP1-Filamin A KD cell lysate Lysates/proteins at 10 µg/Lane. Predicted band size: 281 kDa Observed band size: 281 kDa Exposure time: 20 seconds; ECL: K1801; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (ET1601-3) at 1/2,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. |
|
Fig3:
Immunocytochemistry analysis of HeLa cells labeling Filamin A with Rabbit anti-Filamin A antibody (ET1601-3) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Filamin A antibody (ET1601-3) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
|
Fig4:
Immunocytochemistry analysis of C2C12 cells labeling Filamin A with Rabbit anti-Filamin A antibody (ET1601-3) at 1/500 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Filamin A antibody (ET1601-3) at 1/500 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
|
Fig5:
Immunocytochemistry analysis of C6 cells labeling Filamin A with Rabbit anti-Filamin A antibody (ET1601-3) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Filamin A antibody (ET1601-3) at 1/200 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (HA601187, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. |
|
Fig6: 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. |
|
Fig7: 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. |
|
Fig8:
Flow cytometric analysis of HeLa cells labeling Filamin A. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1601-3, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black). |
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.