AB185204

Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free

RabMAb
Recombinant
KO Validated
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(1 Publication)

Rabbit Recombinant Monoclonal alpha Actinin/ACTN1 antibody. Carrier free. Suitable for WB and reacts with Human, Mouse, Rat samples. Cited in 1 publication.

View Alternative Names

Alpha-actinin-1, Alpha-actinin cytoskeletal isoform, F-actin cross-linking protein, Non-muscle alpha-actinin-1, ACTN1

5 Images

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Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (AB185204)

This data was developed using ab81265, the same antibody clone in a different buffer formulation.

All lanes:

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - Loading Control (<a href='/en-us/products/primary-antibodies/alpha-actinin-actn1-antibody-ep2528y-loading-control-ab81265'>ab81265</a>) at 1/1000 dilution

Lane 1:

NIH 3T3 cell lysate at 10 µg

Lane 2:

human skeletal muscle tissue lysate at 10 µg

Lane 3:

human heart tissue lysate at 10 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

Predicted band size: 103 kDa

Observed band size: 103 kDa

false

Lab

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (AB185204)

This data was developed using ab81265, the same antibody clone in a different buffer formulation.

Lane 1 : Wild-type HAP1 whole cell lysate (20 μg)

Lane 2 : Alpha Actinin knockout HAP1 whole cell lysate (20 μg)

Lane 3 : HeLa whole cell lysate (20 μg)

Lane 4 : NIH 3T3 whole cell lysate (20 μg)

Lanes 1 - 4 : Merged signal (red and green). Green - ab81265 observed at 103 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab81265 was shown to recognize alpha Actinin in wild-type HAP1 cells as signal was lost at the expected MW in alpha Actinin knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and alpha Actinin knockout samples were subjected to SDS-PAGE. ab81265 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - Loading Control (<a href='/en-us/products/primary-antibodies/alpha-actinin-actn1-antibody-ep2528y-loading-control-ab81265'>ab81265</a>)

Predicted band size: 103 kDa

false

Lab

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (AB185204)

This data was developed using the same antibody clone in a different buffer formulation (ab81265).

Lanes 1 - 4 : Merged signal (red and green). Green - ab81265 observed at 115 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

ab81265 was shown to react with ACTN1 in wild-type HAP1 cells in western blot with loss of signal observed in ACTN1 knockout sample. Wild-type and ACTN1 knockout HAP1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab81265 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Lane 1:

Wild-type HAP1 cell lysate at 20 µg

Lane 2:

ACTN1 knockout HAP1 cell lysate at 20 µg

Lane 3:

A431 cell lysate at 20 µg

Lane 4:

Daudi cell lysate at 20 µg

Predicted band size: 103 kDa

Observed band size: 115 kDa

false

Lab

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (AB185204)

This data was developed using ab81265, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

A673 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 103 kDa

Observed band size: 103 kDa

false

Lab

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (AB185204)

This data was developed using ab81265, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Lane 1:

NIH/3T3 whole cell lysate at 20 µg

Lane 2:

C2C12 whole cell lysate at 20 µg

Lane 3:

C6 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 103 kDa

Observed band size: 103 kDa

false

Unconjugated

Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - Loading Control

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EP2528Y

Isotype

IgG

Carrier free

Yes

Reacts with

Mouse, Rat, Human

Applications

applications

Immunogen

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

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Species", "Dilution Info", "Notes"], "tabs": { "all-applications": {"fullname" : "All Applications", "shortname": "All Applications"}, "IP" : {"fullname" : "Immunoprecipitation", "shortname":"IP"}, "FlowCyt" : {"fullname" : "Flow Cytometry", "shortname":"Flow Cyt"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" }, "Mouse": { "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" }, "Rat": { "IP-species-checked": "notRecommended", "IP-species-dilution-info": "", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "" } } }

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Product details

ab185204 is the carrier-free version of ab81265.

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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.

Conjugation ready
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

Compatibility
This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

pH: 7.2 - 7.4 Constituents: PBS

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

Do Not Freeze

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Supplementary information

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

Alpha Actinin-1 also known as ACTN1 or Actn 1 is a cytoskeletal protein that cross-links actin filaments in muscle and non-muscle cells. With a mass of approximately 100 kDa it is an important component of cell structures such as Z-discs in skeletal muscle. Alpha Actinin is expressed in a range of tissues including skeletal cardiac and smooth muscle as well as in various non-muscle cells. Its main role is to organize the actin cytoskeleton providing structural support and enabling cell motility.

Biological function summary

Alpha Actinin-1 helps maintain the structural integrity of cells by anchoring actin filaments and connecting them to integrins or membrane proteins. It often functions as part of a protein complex interacting with other cytoskeletal components and signaling molecules to stabilize cellular structures. In muscle cells it blends with titin and other sarcomeric proteins to assist muscle contraction. Beyond structural roles it partakes in cellular signaling pathways by interacting with signaling proteins and influencing cellular responses to external stimuli.

Pathways

Alpha Actinin-1 is involved in the regulation of the actin cytoskeleton pathway as well as the integrin signaling pathway. In these pathways it collaborates with proteins such as vinculin and paxillin modulating cell adhesion and movement. Its ability to interact with integrin-linked kinases suggests a role in transducing mechanical signals from the extracellular matrix to downstream signaling cascades.

Alpha Actinin-1 has been linked to a spectrum of muscular diseases and certain cancers. Mutations in Alpha Actinin-1 can contribute to hereditary diseases like congenital kidney disorders which affect actin filament organization in podocytes. In oncology altered expression levels of Alpha Actinin-1 can affect cell proliferation impacting tumor growth and metastasis. Its interaction with other proteins like focal adhesion kinase (FAK) underlines its involvement in these pathological conditions influencing cellular adhesion and signal transduction.

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Product protocols

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

Visit the General protocols
Visit the Troubleshooting

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Target data

F-actin cross-linking protein which is thought to anchor actin to a variety of intracellular structures. Association with IGSF8 regulates the immune synapse formation and is required for efficient T-cell activation (PubMed : 22689882).

See full target information ACTN1

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Publications (1)

Recent publications for all applications. Explore the full list and refine your search

eLife 12: PubMed39023520

2024

Netrin signaling mediates survival of dormant epithelial ovarian cancer cells.

Applications

Unspecified application

Species

Unspecified reactive species

Pirunthan Perampalam,James I MacDonald,Komila Zakirova,Daniel T Passos,Sumaiyah Wasif,Yudith Ramos-Valdes,Maeva Hervieu,Patrick Mehlen,Rob Rottapel,Benjamin Gibert,Rohann J M Correa,Trevor G Shepherd,Frederick A Dick

View all publications

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Product promise

We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.

For full details, please see our Terms & Conditions

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

Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (AB185204)

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (AB185204)

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (AB185204)

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (AB185204)

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (AB185204)

Related conjugates and formulations (1)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (1)

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Primary Antibodies

Primary Antibodies

Product promise