Skip to main content

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.


Images

Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (AB108198), expandable thumbnail
  • Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (AB108198), expandable thumbnail
  • Flow Cytometry (Intracellular) - Anti-alpha Actinin 4 antibody [EPR2533(2)] (AB108198), expandable thumbnail
  • Immunocytochemistry/ Immunofluorescence - Anti-alpha Actinin 4 antibody [EPR2533(2)] (AB108198), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Actinin 4 antibody [EPR2533(2)] (AB108198), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
IHC-PIPWBICC/IFFlow Cyt (Intra)
Human
Tested
Tested
Tested
Tested
Tested
Mouse
Tested
Expected
Tested
Expected
Expected
Rat
Tested
Expected
Tested
Expected
Expected

Tested
Tested

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.

Tested
Tested

Species

Human

Dilution info

1/30

Notes

-

Expected
Expected

Species

Rat, Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

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

-

Tested
Tested

Species

Human

Dilution info

1/100 - 1/250

Notes

-

Expected
Expected

Species

Rat, Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Tested
Tested

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.

Expected
Expected

Species

Rat, Mouse

Dilution info

Use at an assay dependent concentration.

Notes

-

Associated Products

Select an associated product type

6 products for Alternative Product

Target data

Function

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).

Alternative names

Recommended products

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.

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR2533(2)

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Stable for 12 months at -20°C

Notes

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:

  • - High batch-to-batch consistency and reproducibility
  • - Improved sensitivity and specificity
  • - Long-term security of supply
  • - Animal-free batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

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.

Biological function summary

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.

Pathways

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.

Associated diseases and disorders

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.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

14 product images

  • Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198), expandable thumbnail

    Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

    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

    Secondary

    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.

  • Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198), expandable thumbnail

    Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

    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

    Secondary

    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 (Intracellular) - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198), expandable thumbnail

    Flow Cytometry (Intracellular) - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

    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/ Immunofluorescence - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

    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) - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

    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) - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

    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) - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

    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.

  • Immunoprecipitation - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198), expandable thumbnail

    Immunoprecipitation - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

    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

  • Immunocytochemistry/ Immunofluorescence - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198), expandable thumbnail

    Immunocytochemistry/ Immunofluorescence - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

    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).

  • Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198), expandable thumbnail

    Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

    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

  • Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198), expandable thumbnail

    Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

    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

    Secondary

    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-alpha Actinin 4 antibody [EPR2533(2)] (ab108198), expandable thumbnail

    Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

    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

    Secondary

    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

  • Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198), expandable thumbnail

    Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

    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

    Secondary

    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 - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198), expandable thumbnail

    Western blot - Anti-alpha Actinin 4 antibody [EPR2533(2)] (ab108198)

    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

    Secondary

    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

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com