AB81265

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

KO Validated
RabMAb
Recombinant
Lab Essentials
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(1 Publication)

Rabbit Recombinant Monoclonal alpha Actinin/ACTN1 antibody. Suitable for WB and reacts with Mouse, Rat, Human 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

4 Images

Lab

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - Loading Control (AB81265)

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 (ab81265)

Predicted band size: 103 kDa

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Lab

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - Loading Control (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:

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - Loading Control (ab81265) at 1/1000 dilution

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

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Lab

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - Loading Control (AB81265)

Blocking and dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - Loading Control (ab81265) at 1/1000 dilution

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] - Loading Control (AB81265)

Blocking and dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - Loading Control (ab81265) at 1/1000 dilution

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

Carrier free

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

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EP2528Y

Isotype

IgG

Carrier free

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": "1/40", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "", "FlowCyt-species-notes": "<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a> - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/500 - 1/1000", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "1/100 - 1/250", "ICCIF-species-notes": "" }, "Mouse": { "IP-species-checked": "notRecommended", "IP-species-dilution-info": "1/40", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "1/40 - 1/100", "FlowCyt-species-notes": "<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a> - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/500 - 1/1000", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "1/100 - 1/250", "ICCIF-species-notes": "" }, "Rat": { "IP-species-checked": "notRecommended", "IP-species-dilution-info": "1/40", "IP-species-notes": "", "FlowCyt-species-checked": "notRecommended", "FlowCyt-species-dilution-info": "1/40 - 1/100", "FlowCyt-species-notes": "<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a> - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.", "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/500 - 1/1000", "WB-species-notes": "", "ICCIF-species-checked": "notRecommended", "ICCIF-species-dilution-info": "1/100 - 1/250", "ICCIF-species-notes": "" } } }

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Product details

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.

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Properties and storage information

Form

Liquid

Purification technique

Affinity purification Protein A

Storage buffer

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

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

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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

Inflammation : PubMed40021543

2025

Inhibition of Cullin3 Neddylation Alleviates Diabetic Retinopathy by Activating Nrf2 Signaling to Combat ROS-Induced Oxidative Stress and Inflammation.

Applications

Unspecified application

Species

Unspecified reactive species

Yueqin Chen,Cong Liu,Jun Tong,Chang He,Xinru Ling,Jinjin Xiang,Chunyan Xue,Genhong Yao,Lingyun Sun,Zhenggao Xie

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] - Loading Control

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - Loading Control (AB81265)

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - Loading Control (AB81265)

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - Loading Control (AB81265)

Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - Loading Control (AB81265)

Related conjugates and formulations (1)

Key facts

Host species

Clonality

Clone number

Isotype

Carrier free

Reacts with

Applications

Immunogen

Alternative Products

Primary Antibodies

Primary Antibodies

Primary Antibodies

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Complementary Products

Secondary Antibodies

Primary Antibodies

Primary Antibodies

Primary Antibodies

Reactivity data

Product details

Properties and storage information

Form

Purification technique

Storage buffer

Shipped at conditions

Appropriate short-term storage duration

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Aliquoting information

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (1)

Product promise