Product Type: | Recombinant Rabbit monoclonal IgG, primary antibodies |
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Species reactivity: | Human, Mouse, Rat |
Applications: | WB, IF-Cell, 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: | A549 cell lysate, HeLa cell lysate, NIH/3T3 cell lysate, Mouse lung tissue lysate, Rat liver tissue lysate, Rat lung tissue lysate, A549, NIH/3T3, mouse liver tissue, rat liver tissue. |
Subcellular location: | Cell junction. Cytoplasm. Nucleus. |
Recommended Dilutions:
WB IF-Cell IHC-P FC IP |
1:2,000 1:100-1:500 1:200 1:1,000 1-2μg/sample |
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 |
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. |
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Fig2:
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 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. Predicted band size: 105 kDa Observed band size: 100 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 (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. |
Fig3:
Immunocytochemistry analysis of A549 cells labeling alpha Actinin 4 with Rabbit anti-alpha Actinin 4 antibody (ET1706-05) 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 (ET1706-05) at 1/100 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. |
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Fig4:
Immunocytochemistry analysis of NIH/3T3 cells labeling alpha Actinin 4 with Rabbit anti-alpha Actinin 4 antibody (ET1706-05) 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 (ET1706-05) at 1/100 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:
Immunohistochemical analysis of paraffin-embedded mouse liver tissue with Rabbit anti-alpha Actinin 4 antibody (ET1706-05) 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 (ET1706-05) 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. |
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Fig6:
Immunohistochemical analysis of paraffin-embedded rat liver tissue with Rabbit anti-alpha Actinin 4 antibody (ET1706-05) 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 (ET1706-05) 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. |
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Fig7:
Flow cytometric analysis of A549 cells labeling alpha Actinin 4. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1706-05, 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). |
Fig8:
Flow cytometric analysis of NIH/3T3 cells labeling alpha Actinin 4. Cells were fixed and permeabilized. Then stained with the primary antibody (ET1706-05, 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). |
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Fig9:
alpha Actinin 4 was immunoprecipitated from 0.2 mg A549 cell lysate with ET1706-05 at 2 µg/10 µl beads. Western blot was performed from the immunoprecipitate using ET1706-05 at 1/1,000 dilution. HRP Conjugated Anti-Rabbit IgG for IP secondary antibody at 1/5,000 dilution was used for 1 hour at room temperature. Lane 1: A549 cell lysate (input) Lane 2: ET1706-05 IP in A549 cell lysate Lane 3: Rabbit IgG instead of ET1706-05 in A549 cell lysate Blocking/Dilution buffer: 5% NFDM/TBST Exposure time: 3 seconds; ECL: K1801 |