WB
Western blot analysis of alpha Actinin on different lysates with Rabbit anti-alpha Actinin antibody at 1/5,000 dilution. Lane 1: A431 cell lysate, Lane 2: NIH/3T3 cell lysate, Lane 3: C6 cell lysate, Lane 4: COS-1 cell lysate, Lane 5: Mouse heart tissue lysate, Lane 6: Rat colon tissue lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 10 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 1/5,000 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 human breast cancer tissue with Rabbit anti-alpha Actinin antibody at 1/1,000 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 1/1,000 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 colon tissue with Rabbit anti-alpha Actinin antibody at 1/1,000 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 1/1,000 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 Siha cells labeling alpha Actinin with Rabbit anti-alpha Actinin antibody at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-alpha Actinin antibody at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. 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 (647) were used as the secondary antibody at 1/1,000 dilution.FC
Flow cytometric analysis of SH-SY5Y cells labeling alpha Actinin. 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).| Product Name | alpha Actinin Rabbit Polyclonal Antibody |
|---|---|
| Antibody Type | Primary Antibodies |
| Product description | Alpha actinins belong to the spectrin gene superfamily which represents a diverse group of cytoskeletal proteins, including the alpha and beta spectrins and dystrophins. Alpha actinin is an actin-binding protein with multiple roles in different cell types. In nonmuscle cells, the cytoskeletal isoform is found along microfilament bundles and adherens-type junctions, where it is involved in binding actin to the membrane. In contrast, skeletal, cardiac, and smooth muscle isoforms are localized to the Z-disc and analogous dense bodies, where they help anchor the myofibrillar actin filaments. This gene encodes a nonmuscle, cytoskeletal, alpha actinin isoform and maps to the same site as the structurally similar erythroid beta spectrin gene. Three transcript variants encoding different isoforms have been found for this gene. |
| Immunogen | Recombinant protein within Human alpha Actinin aa 388-619 / 892. |
| Clonality | Polyclonal |
|---|---|
| Isotype | IgG |
| Host Species | Rabbit |
| Tested Applications | FCICC/IFIHCWB |
| WB:1:5000 IHC:1:1000 ICC/IF:1:200 FC:1:1000 |
|
| Species Reactivity | HumanMonkeyMouseRat |
| Concentration | 1mg/ml |
| Purification | Protein A |
| Alternative Names | actinin 1 smooth muscle antibody
Actinin alpha 1 antibody
actinin alpha 1 antibody ACTN 1 antibody Actn1 antibody ACTN1_HUMAN antibody Alpha Actinin 1 antibody Alpha actinin cytoskeletal isoform antibody Alpha-actinin cytoskeletal isoform antibody Alpha-actinin-1 antibody BDPLT15 antibody F actin cross linking protein antibody F-actin cross-linking protein antibody FLJ40884 antibody FLJ54432 antibody Non muscle alpha actinin 1 antibody Non-muscle alpha-actinin-1 antibody |
|---|---|
| Molecular Weight(MW) | 103 kDa |
| Cellular Localization | Cell junction, Cell membrane, Cell projection, Cytoplasm, Plasma membrane. Cytoskeleton. |
WB
Western blot analysis of alpha Actinin on different lysates with Rabbit anti-alpha Actinin antibody at 1/5,000 dilution. Lane 1: A431 cell lysate, Lane 2: NIH/3T3 cell lysate, Lane 3: C6 cell lysate, Lane 4: COS-1 cell lysate, Lane 5: Mouse heart tissue lysate, Lane 6: Rat colon tissue lysate, Lysates/proteins at 20 µg/Lane. Exposure time: 10 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 1/5,000 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 human breast cancer tissue with Rabbit anti-alpha Actinin antibody at 1/1,000 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 1/1,000 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 colon tissue with Rabbit anti-alpha Actinin antibody at 1/1,000 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 1/1,000 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 Siha cells labeling alpha Actinin with Rabbit anti-alpha Actinin antibody at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-alpha Actinin antibody at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (488) was used as the secondary antibody at 1/1,000 dilution. 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 (647) were used as the secondary antibody at 1/1,000 dilution.
FC
Flow cytometric analysis of SH-SY5Y cells labeling alpha Actinin. 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).| Application Notes | WB:1:5000 IHC:1:1000 ICC/IF:1:200 FC:1:1000 |
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| Form | Liquid |
|---|---|
| Storage Instructions | Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles. |
| Storage Buffer | 1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide. |
