AB200658

Anti-Actin antibody [EPR16875] - Loading Control

RabMAb
Recombinant
Lab Essentials
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(9 Publications)

Rabbit Recombinant Monoclonal alpha skeletal muscle Actin antibody. Suitable for WB, ICC/IF, Flow Cyt (Intra) and reacts with Mouse, Rat, Chicken, Human samples. Cited in 9 publications.

View Alternative Names

ACTA, ACTA1, Alpha-actin-1

7 Images

Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Actin antibody [EPR16875] - Loading Control (AB200658)

Intracellular flow cytometric analysis of 2% paraformaldehyde-fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Actin with ab200658 at 1/100 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat anti rabbit IgG (FITC) at 1/500 dilution was used as the secondary antibody.

ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Actin antibody [EPR16875] - Loading Control (AB200658)

Immunofluorescent analysis of 100% Methanol-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling Actin with ab200658 at 1/1000 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green). Cytoplasm staining on HeLa cell line is observed.

The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

The negative controls are as follows :
-ve control 1 : ab200658 at 1/1000 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

Supplier Data

Western blot - Anti-Actin antibody [EPR16875] - Loading Control (AB200658)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Actin antibody [EPR16875] - Loading Control (ab200658) at 1/2000 dilution

All lanes:

HeLa (Human epithelial cells from cervix adenocarcinoma) cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

Exposure time: 10s

Supplier Data

Western blot - Anti-Actin antibody [EPR16875] - Loading Control (AB200658)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Actin antibody [EPR16875] - Loading Control (ab200658) at 1/10000 dilution

Lane 1:

Jurkat (Human T cell leukemia cells from peripheral blood) cell lysate at 10 µg

Lane 2:

HepG2 (Human liver hepatocellular carcinoma) cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

Exposure time: 3min

Supplier Data

Western blot - Anti-Actin antibody [EPR16875] - Loading Control (AB200658)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 23129404; PMID : 11033016; PMID : 35092032; PMID : 26018190)

The identity of the bands higher than 25 kDa are unknown.

In Western blot, Anti-Actin antibody [EPR16875] - Loading Control (ab200658) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-Interferon gamma antibody [EPR28352-7] (<a href='/en-us/products/primary-antibodies/interferon-gamma-antibody-epr28352-7-ab324874'>ab324874</a>) at 1/1000 dilution

Lane 1:

Untreated mouse spleen cell whole cell lysate at 20 µg

Lane 2:

Mouse spleen cell treated with 50 ng/ml Phorbol-12-myristate-13-acetate (PMA) and 1 μg/ml Ionomycin for 1 hour then add 300ng/ml Brefeldin A for additional 3 hours

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

Observed band size: 17 kDa,42 kDa

false

Exposure time: 15s

Supplier Data

Western blot - Anti-Actin antibody [EPR16875] - Loading Control (AB200658)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Actin antibody [EPR16875] - Loading Control (ab200658) at 1/2000 dilution

Lane 1:

Mouse brain lysate at 10 µg

Lane 2:

Mouse heart lysate at 10 µg

Lane 3:

Mouse kidney lysate at 10 µg

Lane 4:

Mouse spleen lysate at 10 µg

Lane 5:

Rat brain lysate at 10 µg

Lane 6:

Rat heart lysate at 10 µg

Lane 7:

Rat kidney lysate at 10 µg

Lane 8:

C6 (Rat glial tumor cells) cell lysate at 10 µg

Lane 9:

RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

Exposure time: 5s

Supplier Data

Western blot - Anti-Actin antibody [EPR16875] - Loading Control (AB200658)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Actin antibody [EPR16875] - Loading Control (ab200658) at 1/2000 dilution

All lanes:

UMNSAH/DF-1 (Transformed chicken embyronic fibroblast cells) cell lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

false

Exposure time: 5s

578 PE

PE Anti-Actin antibody [EPR16875] - Cytoskeleton Marker
665 Alexa Fluor® 647

Alexa Fluor® 647 Anti-Actin antibody [EPR16875] - Cytoskeleton Marker
Carrier free

Anti-Actin antibody [EPR16875] - BSA and Azide free
660 APC

APC Anti-Actin antibody [EPR16875] - Cytoskeleton Marker
519 Alexa Fluor® 488

Alexa Fluor® 488 Anti-Actin antibody [EPR16875] - Cytoskeleton Marker

Key facts

Host species

Rabbit

Clonality

Monoclonal

Clone number

EPR16875

Isotype

IgG

Carrier free

Reacts with

Mouse, Rat, Chicken, Human

Applications

WB, Flow Cyt (Intra), ICC/IF

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"}, "WB" : {"fullname" : "Western blot", "shortname":"WB"}, "ICCIF" : {"fullname" : "Immunocytochemistry/ Immunofluorescence", "shortname":"ICC/IF"}, "FlowCytIntra" : {"fullname" : "Flow Cytometry (Intracellular)", "shortname":"Flow Cyt (Intra)"} }, "product-promise": { "all": "all", "testedAndGuaranteed": "tested", "guaranteed": "expected", "predicted": "predicted", "notRecommended": "not-recommended" } }, "values": { "Human": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/2000", "WB-species-notes": "", "ICCIF-species-checked": "testedAndGuaranteed", "ICCIF-species-dilution-info": "1/1000", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "testedAndGuaranteed", "FlowCytIntra-species-dilution-info": "1/100", "FlowCytIntra-species-notes": "" }, "Mouse": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/2000", "WB-species-notes": "", "ICCIF-species-checked": "guaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "guaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" }, "Rat": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/2000", "WB-species-notes": "", "ICCIF-species-checked": "guaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "guaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-species-notes": "" }, "Chicken": { "WB-species-checked": "testedAndGuaranteed", "WB-species-dilution-info": "1/2000", "WB-species-notes": "", "ICCIF-species-checked": "guaranteed", "ICCIF-species-dilution-info": "", "ICCIF-species-notes": "", "FlowCytIntra-species-checked": "guaranteed", "FlowCytIntra-species-dilution-info": "", "FlowCytIntra-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

Avoid freeze / thaw cycle

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Supplementary information

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

Actin also known as globular (G-actin) or filamentous (F-actin) actin plays a central role in cell structure and movement. This protein has a molecular weight of approximately 42 kDa and resides abundantly in eukaryotic cells especially in muscle and cytoskeletal structures. It comes in several isoforms with varying expression profiles across tissues. Actin can undergo polymerization from its G-actin monomers into F-actin filaments a process that is reversible through depolymerization allowing for dynamic cellular functions.

Biological function summary

Actin contributes to various cellular processes by forming the cytoskeleton which provides mechanical support and determines cell shape. Actin also facilitates cell motility division and intracellular transport through rapid polymerization and depolymerization cycles. Within cells actin associates with other proteins to form complexes such as the Arp2/3 complex which assists actin in the branching of filaments critical for pushing forward the cell's leading edge during movement. Techniques like actin immunofluorescence help visualize actin filaments within cells revealing its extensive network.

Pathways

Actin plays an integral role in pathways like cell movement and signal transduction. The Rho family of GTPases regulates actin cytoskeleton rearrangements influencing cell shape and migration. Actin function interactions involve proteins like myosin forming actomyosin complexes essential for muscle contraction and other cell motility activities. Actin polymerization and depolymerization cycles are key to the dynamic regulation within these pathways ensuring adequate cellular responses to environmental signals.

Mutations or misregulation of actin and its associated pathways link to conditions such as cardiomyopathies and cancer metastasis. In familial cardiomyopathy actin mutations disrupt normal cardiac muscle contraction. In cancer altered actin polymerization and depolymerization enable invasive cell migration facilitating metastasis. Proteins like myosin and tropomyosin associate with actin in these pathologies illustrating the impact of actin dynamics on disease progression and highlighting actin as a therapeutic target. Anti-actin antibodies can be used in research and diagnostics to better understand these disorders.

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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

Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.

See full target information ACTA1

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Publications (9)

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

NPJ Regenerative medicine 8:25 PubMed37217533

2023

A superior extracellular matrix binding motif to enhance the regenerative activity and safety of therapeutic proteins.

Applications

Unspecified application

Species

Unspecified reactive species

Yasmin K Alshoubaki,Yen-Zhen Lu,Julien M D Legrand,Rezvan Karami,Mathilde Fossat,Ekaterina Salimova,Ziad Julier,Mikaël M Martino

Open medicine (Warsaw, Poland) 18:20230700 PubMed37215053

2023

NECAB3 promotes the migration and invasion of liver cancer cells through HIF-1α/RIT1 signaling pathway.

Applications

Unspecified application

Species

Unspecified reactive species

Yicheng Tian,Longjiang Shao,Qi Wang,Gan Ru,Chuntao Zhai,Lihui Zhou

Cell biology international 47:1081-1091 PubMed36861686

2023

COL11A1-driven positive feedback loop modulates fibroblast transformation and activates pancreatic cancer progression.

Applications

Unspecified application

Species

Unspecified reactive species

Defeng Sun,Tu Dai,Yuan Ji,Weibo Shen,Wuyang Bian

Biomaterials research 27:3 PubMed36647161

2023

Engineered exosomes enriched in netrin-1 modRNA promote axonal growth in spinal cord injury by attenuating inflammation and pyroptosis.

Applications

Unspecified application

Species

Unspecified reactive species

Xiao Lu,Guangyu Xu,Zhidi Lin,Fei Zou,Siyang Liu,Yuxuan Zhang,Wei Fu,Jianyuan Jiang,Xiaosheng Ma,Jian Song

Molecular therapy : the journal of the American Society of Gene Therapy 31:552-568 PubMed36245126

2022

HCP5 prevents ubiquitination-mediated UTP3 degradation to inhibit apoptosis by activating c-Myc transcriptional activity.

Applications

Unspecified application

Species

Unspecified reactive species

Yabing Nan,Qingyu Luo,Xiaowei Wu,Wan Chang,Pengfei Zhao,Shi Liu,Zhihua Liu

The Journal of biological chemistry 298:101803 PubMed35257744

2022

Structural and biophysical characterization of transcription factor HNF-1A as a tool to study MODY3 diabetes variants.

Applications

Unspecified application

Species

Unspecified reactive species

Laura Kind,Arne Raasakka,Janne Molnes,Ingvild Aukrust,Lise Bjørkhaug,Pål Rasmus Njølstad,Petri Kursula,Thomas Arnesen

Journal of biochemical and molecular toxicology 36:e22981 PubMed34984768

2022

Hesperidin delays cell cycle progression into the G0/G1 phase via suspension of MAPK signaling pathway in intrahepatic cholangiocarcinoma.

Applications

Unspecified application

Species

Unspecified reactive species

Jie Deng,Li Liu,Li Li,Jianhai Sun,Fei Yan

Environmental toxicology 36:1052-1060 PubMed33475233

2021

Hsa_circ_0001073 targets miR-626/LIFR axis to inhibit lung cancer progression.

Applications

Unspecified application

Species

Unspecified reactive species

Qun Liu,Gang Cao,Yufeng Wan,Chuanqin Xu,Yuanqiang He,Gang Li

Oncology letters 20:208 PubMed32963614

2020

Potential biomarkers Ang II/AT1R and S1P/S1PR1 predict the prognosis of hepatocellular carcinoma.

Applications

Unspecified application

Species

Unspecified reactive species

Yuanyuan Ji,Haiyan Chen,Wei Gow,Li Ma,Yaofeng Jin,Bo Hui,Zhengan Yang,Zhidong Wang

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-Actin antibody [EPR16875] - Loading Control

Flow Cytometry (Intracellular) - Anti-Actin antibody [EPR16875] - Loading Control (AB200658)

Immunocytochemistry/ Immunofluorescence - Anti-Actin antibody [EPR16875] - Loading Control (AB200658)

Western blot - Anti-Actin antibody [EPR16875] - Loading Control (AB200658)

Western blot - Anti-Actin antibody [EPR16875] - Loading Control (AB200658)

Western blot - Anti-Actin antibody [EPR16875] - Loading Control (AB200658)

Western blot - Anti-Actin antibody [EPR16875] - Loading Control (AB200658)

Western blot - Anti-Actin antibody [EPR16875] - Loading Control (AB200658)

Related conjugates and formulations (5)

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 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 (9)

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