Rabbit Recombinant Monoclonal alpha Actinin 4 antibody. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Rat, Mouse, Human samples. Cited in 32 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
IHC-P | IP | WB | ICC/IF | Flow Cyt (Intra) | |
---|---|---|---|---|---|
Human | Tested | Tested | Tested | Tested | Tested |
Mouse | Tested | Expected | Tested | Expected | Expected |
Rat | Tested | Expected | Tested | Expected | Expected |
Species | Dilution info | Notes |
---|---|---|
Species Rat | Dilution info 1/100 - 1/250 | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species Mouse | Dilution info 1/100 - 1/250 | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species Human | Dilution info 1/100 - 1/250 | Notes Perform heat-mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/30 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Mouse | Dilution info 1/1000 - 1/10000 | Notes - |
Species Rat | Dilution info 1/1000 - 1/10000 | Notes - |
Species Human | Dilution info 1/1000 - 1/10000 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info 1/100 - 1/250 | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
Species | Dilution info | Notes |
---|---|---|
Species Human | Dilution info - | Notes Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
Species | Dilution info | Notes |
---|---|---|
Species Rat, Mouse | Dilution info Use at an assay dependent concentration. | Notes - |
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F-actin cross-linking protein which is thought to anchor actin to a variety of intracellular structures. This is a bundling protein (Probable). Probably involved in vesicular trafficking via its association with the CART complex. The CART complex is necessary for efficient transferrin receptor recycling but not for EGFR degradation (PubMed:15772161). Involved in tight junction assembly in epithelial cells probably through interaction with MICALL2. Links MICALL2 to the actin cytoskeleton and recruits it to the tight junctions (By similarity). May also function as a transcriptional coactivator, stimulating transcription mediated by the nuclear hormone receptors PPARG and RARA (PubMed:22351778). Association with IGSF8 regulates the immune synapse formation and is required for efficient T-cell activation (PubMed:22689882).
Alpha-actinin-4, Non-muscle alpha-actinin 4, ACTN4
Rabbit Recombinant Monoclonal alpha Actinin 4 antibody. Suitable for IHC-P, IP, WB, ICC/IF, Flow Cyt (Intra) and reacts with Rat, Mouse, Human samples. Cited in 32 publications.
IgG
Rabbit
pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Liquid
Monoclonal
EPR2533(2)
Affinity purification Protein A
Blue Ice
1-2 weeks
+4°C
-20°C
Upon delivery aliquot
Stable for 12 months at -20°C
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
This product is a recombinant monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
This supplementary information is collated from multiple sources and compiled automatically.
Alpha Actinin 4 sometimes referred to as ACTN4 or alpha-actinin is a protein with a mass of approximately 100 kDa. It functions as an actin-binding protein particularly involved in the crosslinking of actin filaments. This protein is expressed widely with high levels found in muscle kidney and brain tissues. Alpha-actinin 4 also plays important roles in cellular structure and integrity by binding actin filaments in various cellular contexts.
The role of alpha-actinin 4 extends to maintaining cytoskeletal structure and cell motility. It forms part of the larger actin-binding complex that supports cellular adhesion and mechanical stability. By linking actin filaments it allows cells to withstand mechanical stress and maintain their shape. Additionally alpha-actinin 4 contributes to cellular processes like cytokinesis and signal transduction by serving as a scaffold for signaling proteins.
Alpha-actinin 4 interacts with diverse cellular signaling mechanisms. Key pathways include the actin cytoskeleton signaling and integrin-mediated cell adhesion. In these pathways alpha-actinin 4 works alongside proteins such as integrins and vinculin facilitating communication between the extracellular matrix and the cytoskeleton. These interactions are critical for cellular responses to environmental stimuli and adaptation during movement or development.
Alpha-actinin 4 has notable impacts on conditions like focal segmental glomerulosclerosis (FSGS) and cancer. In FSGS mutation or dysregulation of alpha-actinin 4 contributes to podocyte dysfunction leading to proteinuria and kidney damage. Its role in cancer often involves altered expression influencing cell migration and invasion. The interaction with other proteins like nephrin in FSGS or E-cadherin in cancer highlights the significance of alpha-actinin 4 in pathogenic processes.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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All lanes: Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198) at 1/10000 dilution
All lanes: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 15 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 105 kDa
Observed band size: 105 kDa
This data was developed using ab108198, the same antibody clone in a different buffer formulation.
All lanes: Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198) at 1/1000 dilution
Lane 1: Human skeletal muscle lysate at 20 µg
Lane 2: A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg
Lane 3: Mouse brain lysate at 20 µg
Lane 4: Rat brain lysate at 20 µg
Lane 5: Rat heart lysate at 20 µg
All lanes: Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution
Predicted band size: 105 kDa
Observed band size: 105 kDa
Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling alpha Actinin 4 with Purified ab108198 at 1:50 dilution (10 µg/ml) (Red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody was used at 1:2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (Blue).
Immunocytochemistry analysis of MCF7 (Human breast adenocarcinoma epithelial cell) cells labeling alpha Actinin 4 with Purified ab108198 at 1:250 dilution (2.0 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat liver tissue sections labeling alpha Actinin 4 with Purified ab108198 at 1:150 dilution (3.29 µg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse liver tissue sections labeling alpha Actinin 4 with Purified ab108198 at 1:150 dilution (3.29 µg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling alpha Actinin 4 with Purified ab108198 at 1:150 dilution (3.29 µg/mL). Heat mediated antigen retrieval was performed using Perform heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). Tissue was counterstained with Hematoxylin. ImmunoHistoProbe one step HRP Polymer (ready to use) secondary antibody was used. PBS instead of the primary antibody was used as the negative control.
Purified ab108198 at 1/50 dilution (2μg) immunoprecipitating alpha Actinin 4 in HeLa whole cell lysate.
Lane 1 (input): HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate 10μg
Lane 2 (+): ab108198 + HeLa whole cell lysate.
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab108198 in HeLa whole cell lysate.
VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) (1/1000 dilution) was used for Western blotting.
Blocking Buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM/TBST.
Observed band size: 105 kDa
All lanes: Immunoprecipitation - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)
Predicted band size: 105 kDa
ab108198 staining ACTN4 in wild-type HAP1 cells (top panel) and ACTN4 knockout HAP1 cells (bottom panel). The cells were fixed with PFA (10min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab108198 at 1/100 dilution and Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291 (Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
Lane 1: Wild-type HAP1 whole cell lysate (20 μg)
Lane 2: ACTN4 (alpha Actinin 4) knockout HAP1 whole cell lysate (20 μg)
Lane 3: HeLa whole cell lysate (20 μg)
Lane 4: MCF7 whole cell lysate (20 μg)
Lanes 1 - 4: Merged signal (red and green). Green - ab108198 observed at 105 kDa. Red - loading control, Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484, observed at 37 kDa.
ab108198 was shown to specifically react with alpha Actinin 4 in wild-type HAP1 cells as signal was lost in ACTN4 (alpha Actinin 4) knockout cells. Wild-type and ACTN4 (alpha Actinin 4) knockout samples were subjected to SDS-PAGE. ab108198 and Anti-GAPDH antibody [mAbcam 9484] - Loading Control ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)
Predicted band size: 105 kDa
Anti-CD38 antibody [5C5C3] - Extracellular domain staining at 1/500 dilution, shown in green; Rabbit anti-ACTN4 [EPR2533(2)] (ab108198) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-CD38 antibody [5C5C3] - Extracellular domain ab204940 was shown to bind specifically to CD38. A band was observed at 42 kDa in wild-type A549 cell lysates with no signal observed at this size in CD38 knockout cell line. To generate this image, wild-type and CD38 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-CD38 antibody [5C5C3] - Extracellular domain (Anti-CD38 antibody [5C5C3] - Extracellular domain ab204940) at 1/500 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: CD38 knockout A549 cell lysate at 20 µg
Lane 3: Daudi cell lysate at 20 µg
Lane 4: HCT 116 cell lysate at 20 µg
All lanes: Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 34 kDa
Observed band size: 42 kDa
Western blot: Anti-LIPA antibody [1F9] (Anti-Lysosomal acid lipase/LAL antibody [1F9] ab219113) staining at 1/1000 dilution, shown in green; Rabbit anti-ACTN4 [EPR2533(2)] (ab108198) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, Anti-Lysosomal acid lipase/LAL antibody [1F9] ab219113 was shown to bind specifically to LIPA. A band was observed at 54 kDa in wild-type A549 cell lysates with no signal observed at this size in LIPA knockout cell line. To generate this image, wild-type and LIPA knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse IgG H&L 800CW and Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution.
All lanes: Western blot - Anti-Lysosomal acid lipase/LAL antibody [1F9] (Anti-Lysosomal acid lipase/LAL antibody [1F9] ab219113) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: LIPA knockout A549 cell lysate at 20 µg
Lanes 1 - 2: Goat anti-Mouse IgG H&L 800CW at 1/20000 dilution
Lanes 1 - 2: Goat anti-Rabbit IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Observed band size: 54 kDa
Negative control: 293T (PMID: 20444890).
The expression profile is consistent with what has been described in the literature (PMID: 20444890).
This blot was produced using a 4-12% Bis-Tris gel and MOPS buffer. The gel was run at 200V for 54 minutes and then transferred to a Nitrocellulose membrane at 25V for 10 minutes. The membrane was blocked for an hour using 3% milk and incubated overnight at 4°C with Mouse monoclonal [CM2B4] to Polyoma virus, Large T antigen ( Anti-Polyoma virus, Large T antigen antibody [CM2B4] ab307450) and Rabbit anti-ACTN4 (loading control, ab108198) at a 1/1000 and 1/20 000 dilution, respectively. Antibody binding was detected using Goat Anti-Mouse IgG H&L (IRDye® 800CW, green) and Goat Anti-Rabbit IgG H&L (IRDye® 680RD, red) secondary antibodies at a 1/20 000 dilution (1h at room temperature) before imaging.
Blocking/Dilution buffer: 3% NFDM/TBST.
All lanes: Western blot - Anti-Polyoma virus, Large T antigen antibody [CM2B4] (Anti-Polyoma virus, Large T antigen antibody [CM2B4] ab307450) at 1/1000 dilution
Lane 1: MKL-1 whole cell lysate at 20 µg
Lane 2: 293T whole cell lysate at 20 µg
All lanes: Western blot - Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) at 1/20000 dilution
Observed band size: 48 kDa
Western blot: Mouse Monoclonal[10/Fibronectin] to Fibronectin Anti-Fibronectin antibody [10/Fibronectin] ab281575 staining at 1/500 dilution, shown in green; Rabbit anti-ACTN4 (ab108198) loading control staining at 1/20,000 dilution, shown in magenta. A band was observed at 238-268 kDa in Wild-type MCF7 cell lysates with no signal observed at this size in FN1 knockout MCF7 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % Milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20,000 dilution.
All lanes: Western blot - Anti-Fibronectin antibody [10/Fibronectin] (Anti-Fibronectin antibody [10/Fibronectin] ab281575) at 1/500 dilution
Lane 1: Wild-type MCF7 at 20 µg
Lane 2: Western blot - Human FN1 knockout MCF7 cell line (Human FN1 knockout MCF7 cell line ab286324) at 20 µg
Lane 3: HepG2 at 20 µg
Lane 4: PC-3 at 20 µg
All lanes: Goat anti-Mouse 800CW & Goat anti-Rabbit 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 272 kDa
Observed band size: 238 kDa, 268 kDa
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