alpha Actinin Mouse Monoclonal Antibody [A1G2]
cat.: EM1901-52
Product Type: Mouse monoclonal IgG1, primary antibodies
Species reactivity: Human, Rat
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: A1G2
Form: Liquid
Storage condition: 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.
Concentration: 2ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 103 kDa
Isotype: IgG1
Immunogen: Recombinant protein within Human ACTN1 aa 388-619 / 892.
Positive control: HeLa cell lysate, A431 cell lysate, A549 cell lysate, MCF7 cell lysate, PC-12 cell lysate, C6 cell lysate, A431, SiHa, JAR, human lung tissue, human liver carcinoma tissue, human skin tissue, human breast tissue, human breast carcinoma tissue, human kidney tissue.
Subcellular location: Cell membrane, cytoskeleton, Z line, cell junction, ruffle.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:1,000
1:50-1:100
1:50-1:500
1:50-1:100
Uniprot #: SwissProt: P12814 Human | Q9Z1P2 Rat
Alternative names: actinin 1 smooth muscle Actinin alpha 1 actinin, alpha 1 ACTN 1 Actn1 ACTN1_HUMAN Alpha Actinin 1 Alpha actinin cytoskeletal isoform Alpha-actinin cytoskeletal isoform Alpha-actinin-1 BDPLT15 F actin cross linking protein F-actin cross-linking protein FLJ40884 FLJ54432 Non muscle alpha actinin 1 Non-muscle alpha-actinin-1
Images
EM1901-52_1.jpg Fig1: Western blot analysis of alpha Actinin on different lysates with Mouse anti-alpha Actinin antibody (EM1901-52) at 1/1,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: A431 cell lysate
Lane 3: A549 cell lysate
Lane 4: MCF7 cell lysate
Lane 5: PC-12 cell lysate
Lane 6: C6 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 103 kDa
Observed band size: 103 kDa

Exposure time: 1 minute;

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 (EM1901-52) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
EM1901-52_2.jpg Fig2: Immunocytochemistry analysis of A431 cells labeling alpha Actinin with Mouse anti-alpha Actinin antibody (EM1901-52) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-alpha Actinin antibody (EM1901-52) at 1/50 dilution in 2% BSA overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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.
EM1901-52_3.jpg Fig3: Immunocytochemistry analysis of SiHa cells labeling alpha Actinin with Mouse anti-alpha Actinin antibody (EM1901-52) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-alpha Actinin antibody (EM1901-52) at 1/50 dilution in 2% BSA overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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.
EM1901-52_4.jpg Fig4: Immunocytochemistry analysis of JAR cells labeling alpha Actinin with Mouse anti-alpha Actinin antibody (EM1901-52) at 1/50 dilution.

Cells were fixed in 4% paraformaldehyde for 30 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 2% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-alpha Actinin antibody (EM1901-52) at 1/50 dilution in 2% BSA overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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.
EM1901-52_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human lung tissue using anti-alpha Actinin 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 (EM1901-52, 1/50) 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.
EM1901-52_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human liver carcinoma tissue using anti-alpha Actinin 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 (EM1901-52, 1/50) 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.
EM1901-52_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human skin tissue using anti-alpha Actinin 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 (EM1901-52, 1/50) 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.
EM1901-52_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human breast tissue using anti-alpha Actinin 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 (EM1901-52, 1/50) 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.
EM1901-52_9.jpg Fig9: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-alpha Actinin 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 (EM1901-52, 1/50) 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.
EM1901-52_10.jpg Fig10: Immunohistochemical analysis of paraffin-embedded human kidney tissue using anti-alpha Actinin 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 (EM1901-52, 1/200) 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.
EM1901-52_11.jpg Fig11: Flow cytometric analysis of alpha Actinin was done on A431 cells. The cells were fixed, permeabilized and stained with the primary antibody (EM1901-52, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
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