WB
Western blot analysis of alpha Actinin 4 on different lysates with Rabbit anti-alpha Actinin 4 antibody at 1/2,000 dilution. Lane 1: A549 cell lysate, Lane 2: HeLa cell lysate, Lane 3: NIH/3T3 cell lysate, Lane 4: Mouse lung tissue lysate, Lane 5: Rat liver tissue lysate, Lane 6: Rat lung tissue lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 20 seconds; 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.IHC
Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-alpha Actinin 4 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour 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.IHC
Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-alpha Actinin 4 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour 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.ICC/IF
Immunocytochemistry analysis of A549 cells labeling alpha Actinin 4 with Rabbit anti-alpha Actinin 4 antibody at 1/100 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-alpha Actinin 4 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of A549 cells labeling alpha Actinin 4. Cells were fixed and permeabilized. Then stained with the primary antibody (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).IP
alpha Actinin 4 was immunoprecipitated from 0.2 mg A549 cell lysate with Rabbit anti-alpha Actinin 4 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-alpha Actinin 4 antibody at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: A549 cell lysate (input), Lane 2: Rabbit anti-alpha Actinin 4 antibody IP in A549 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-alpha Actinin 4 antibody in A549 cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 3 seconds.| Product Name | alpha Actinin 4 Recombinant Rabbit Monoclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | The spectrin gene family encodes a diverse group of cytoskeletal proteins that include spectrins, dystrophins and α-actinins. There are four tissue-specific α-actinins, namely α-actinin-1, α-actinin-2, α-actinin-3 and α-actinin-4, which are localized to muscle and non-muscle cells, including skeletal, cardiac and smooth muscle cells, as well as within the cytoskeleton. Each α-actinin protein contains one Actin-binding domain, two calponin-homology domains, two EF-hand domains and four spectrin repeats, through which they function as bundling proteins that can cross-link F-Actin, thus anchoring Actin to a variety of intracellular structures. Defects in the gene encoding α-actinin-4 are the cause of focal segmental glomerulosclerosis 1 (FSGS1), a common renal lesion characterized by decreasing kidney function and, ultimately, renal failure. are actually sensitive to the Profilin proteins in these foods. |
| Immunogen | Synthetic peptide within Human alpha Actinin 4 aa 1-50 / 911. |
| Clonality | Monoclonal |
|---|---|
| Isotype | IgG |
| Host Species | Recombinant rabbit |
| Tested Applications | FCICC/IFIHCIPWB |
| WB:1:2000 IHC:1:200 ICC/IF:1:100-1:500 FC:1:1000 IP: 1-2μg/sample |
|
| Species Reactivity | HumanMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | actinin 4 antibody Actinin alpha 4 antibody actinin4 antibody ACTN 4 antibody ACTN4 antibody ACTN4_HUMAN antibody Alpha-actinin-4 antibody DKFZp686K23158 antibody F actin cross linking protein antibody F-actin cross-linking protein antibody Focal segmental glomerulosclerosis 1 antibody FSGS 1 antibody FSGS antibody FSGS1 antibody Non muscle alpha actinin 4 antibody Non-muscle alpha-actinin 4 antibody |
|---|---|
| Molecular Weight(MW) | 105 kDa(Observed band size: 100 kDa) |
| Cellular Localization | Cell junction. Cytoplasm. Nucleus. |
WB
Western blot analysis of alpha Actinin 4 on different lysates with Rabbit anti-alpha Actinin 4 antibody at 1/2,000 dilution. Lane 1: A549 cell lysate, Lane 2: HeLa cell lysate, Lane 3: NIH/3T3 cell lysate, Lane 4: Mouse lung tissue lysate, Lane 5: Rat liver tissue lysate, Lane 6: Rat lung tissue lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 20 seconds; 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 at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody at 1/50,000 dilution was used for 1 hour at room temperature.
IHC
Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-alpha Actinin 4 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour 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.
IHC
Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-alpha Actinin 4 antibody at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/200 dilution for 1 hour 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.
ICC/IF
Immunocytochemistry analysis of A549 cells labeling alpha Actinin 4 with Rabbit anti-alpha Actinin 4 antibody at 1/100 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-alpha Actinin 4 antibody at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) 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 (red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (594) was used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of A549 cells labeling alpha Actinin 4. Cells were fixed and permeabilized. Then stained with the primary antibody (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 488 conjugate-Goat anti-Rabbit IgG Secondary antibody at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
IP
alpha Actinin 4 was immunoprecipitated from 0.2 mg A549 cell lysate with Rabbit anti-alpha Actinin 4 antibody at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using Rabbit anti-alpha Actinin 4 antibody at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP Nano-secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: A549 cell lysate (input), Lane 2: Rabbit anti-alpha Actinin 4 antibody IP in A549 cell lysate, Lane 3: Rabbit IgG instead of Rabbit anti-alpha Actinin 4 antibody in A549 cell lysate, Blocking/Dilution buffer: 5% NFDM/TBST, Exposure time: 3 seconds.| Application Notes | WB:1:2000 IHC:1:200 ICC/IF:1:100-1:500 FC:1:1000 IP: 1-2μg/sample |
|---|
| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*TBS (pH7.4), 1%BSA, 40%Glycerol. Preservative: 0.05% Sodium Azide. |
