alpha Actinin 4 Recombinant Rabbit Monoclonal Antibody [JU20-23]
cat.: ET1706-05
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, IF-Tissue, IP, IHC-P, FC
Clonality: Monoclonal
Clone number: JU20-23
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: 105 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human alpha Actinin 4 aa 1-50 / 911.
Positive control: Hela cell lysate, PC-12 cell lysate, NIH-3T3 cell lysate, rat liver tissue lysate, A431 cell lysate, HepG2 cell lysate, A549, Hela, LOVO, human colon carcinoma tissue, mouse colon tissue,A549 .
Subcellular location: Cell junction. Cytoplasm. Nucleus.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
  IP
1:500-1:2,000
1:100-1:500
1:50-1:200
1:50-1:100
1:10-1:50
Uniprot #: SwissProt: O43707 Human | P57780 Mouse | Q9QXQ0 Rat
Alternative names: actinin 4 Actinin alpha 4 actinin4 ACTN 4 ACTN4 ACTN4_HUMAN Alpha-actinin-4 DKFZp686K23158 F actin cross linking protein F-actin cross-linking protein Focal segmental glomerulosclerosis 1 FSGS 1 FSGS FSGS1 Non muscle alpha actinin 4 Non-muscle alpha-actinin 4
Images
ET1706-05_1.jpg Fig1: Western blot analysis of alpha Actinin 4 on different lysates with Rabbit anti-alpha Actinin 4 antibody (ET1706-05) at 1/2,000 dilution.

Lane 1: A549-si NT cell lysate
Lane 2: A549-si alpha Actinin 4 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 105 kDa
Observed band size: 100 kDa

Exposure time: 30 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 (ET1706-05) 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.
ET1706-05_2.jpg Fig2: Western blot analysis of alpha Actinin 4 on different lysates with Rabbit anti-alpha Actinin 4 antibody (ET1706-05) at 1/500 dilution.

Lane 1: Hela cell lysates
Lane 2: PC-12 cell lysates
Lane 3: NIH-3T3 cell lysates
Lane 4: Rat liver tissue lysates(20 µg/Lane)
Lane 5: A431 cell lysates
Lane 6: HepG2 cell lysates

Lysates/proteins at 10 µg/Lane.

Predicted band size: 105 kDa
Observed band size: 100 kDa

Exposure time: 2 minutes;

8% 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 (ET1706-05) at 1/500 dilution was used in 5% NFDM/TBST 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.
ET1706-05_3.jpg Fig3: Immunocytochemistry analysis of A549 cells labeling alpha Actinin 4 with Rabbit anti-alpha Actinin 4 antibody (ET1706-05) at 1/50 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 4 antibody (ET1706-05) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET1706-05_4.jpg Fig4: Immunocytochemistry analysis of Hela cells labeling alpha Actinin 4 with Rabbit anti-alpha Actinin 4 antibody (ET1706-05) at 1/50 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 4 antibody (ET1706-05) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution Nuclear DNA was labelled in blue with DAPI.
ET1706-05_5.jpg Fig5: Immunocytochemistry analysis of LOVO cells labeling alpha Actinin 4 with Rabbit anti-alpha Actinin 4 antibody (ET1706-05) at 1/50 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 4 antibody (ET1706-05) at 1/50 dilution in 2% negative goat serum overnight at 4 ℃. Alexa Fluor®488 conjugate-Goat anti-Rabbit IgG was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.
ET1706-05_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Rabbit anti-alpha Actinin 4 antibody (ET1706-05) at 1/50 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 (ET1706-05) at 1/50 dilution for 0.5 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.
ET1706-05_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded mouse colon tissue with Rabbit anti-alpha Actinin 4 antibody (ET1706-05) at 1/50 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 (ET1706-05) at 1/50 dilution for 0.5 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.
ET1706-05_8.jpg Fig8: Flow cytometric analysis of A549 cells with alpha Actinin 4 antibody at 1/100 dilution (red) compared with an unlabelled 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”.