beta-Actin Antibody

Images

 
Immunocytochemistry/ Immunofluorescence: beta-Actin Antibody [NB600-503] - Cultured pig trabecular meshwork cells stained with beta-Actin antibody at a dilution of 1:500 followed by an anti-rabbit Alexa Fluor488 at ...read more
Western Blot: beta-Actin Antibody [NB600-503] - Rabbit polyclonal at 1/5000.
Simple Western: beta-Actin Antibody [NB600-503] - Simple Western lane view shows a specific band for Beta Actin in 0.1 mg/ml of HeLa lysate. This experiment was performed under reducing conditions using the 12-230 kDa ...read more
Flow Cytometry: beta-Actin Antibody [NB600-503] - An intracellular stain was performed on HeLa cells with beta-actin Antibody NB600-503AF488 (blue) and a matched isotype control (orange). Cells were fixed with 4% PFA ...read more
Western Blot: beta-Actin Antibody [NB600-503] - Western blot analysis of Jurkat, COS, and CHO cell lysate using beta actin antibody at 1:100.
Immunohistochemistry-Paraffin: beta-Actin Antibody [NB600-503] - Analysis of Beta Actin in mouse epidermis using DAB with hematoxylin counterstain.
Immunocytochemistry/ Immunofluorescence: beta-Actin Antibody [NB600-503] - IF Confocal analysis of HeLa cells using Beta Actin antibody (NB600-503, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as ...read more
Flow Cytometry: beta-Actin Antibody [NB600-503] - Analysis of HeLa cells using mouse Monoclonal beta-Actin antibody (Orange) and Isotype control Antibody (Blue).

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Av, ChHa, Fi, Pm, RbSpecies Glossary
Applications WB, Simple Western, EIA, Flow, ICC/IF, IHC, IHC-P, IP, PLA, B/N
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
1.0 mg/ml

beta-Actin Antibody Summary

Immunogen
This beta-Actin Antibody was made from a synthetic peptide made to an N-terminal region of human Beta Actin. [UniProt P60709]
Localization
Cytoplasmic
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
ACTB
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Western Blot 1:100-1:2000
  • Simple Western 1:12.5
  • Enzyme Immunoassay
  • Flow Cytometry 1:10-1:1000
  • Immunocytochemistry/Immunofluorescence
  • Immunohistochemistry 1:100
  • Immunohistochemistry-Paraffin 1:100
  • Immunoprecipitation 1:10-1:500
  • Proximity Ligation Assay
  • Block/Neutralize
Application Notes
Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended. In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.

Use in Immunocytochemistry/Immunofluorescence reported in verified customer review. Use in Blocking/Neutralizing and use in Proximity Ligation Assay reported in scientific literature (PMID: 17056548 and 24740405 respectively).
Theoretical MW
42 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Control
NIH 3T3 Whole Cell Lysate
HeLa Whole Cell Lysate
beta-Actin Knockout 293T Cell Lysate
Reviewed Applications
Read 4 Reviews rated 5
using
NB600-503 in the following applications:

Publications
Read Publications using
NB600-503 in the following applications:

  • B/N
    1 publication
  • IHC
    1 publication
  • 2 publications
  • PLA
    1 publication
  • WB
    54 publications

Reactivity Notes

Rabbit reactivity and Fish reactivity reported in scientific literature (PMID: 23813946 and 25842206 respectively). Expected to cross-react with a wide range of species due to sequence identity.

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.
Buffer
Tris-Citrate/Phosphate (pH 7.0 - 8.0)
Preservative
0.09% Sodium Azide
Concentration
1.0 mg/ml
Purity
Immunogen affinity purified

Alternate Names for beta-Actin Antibody

  • ACTB
  • actin, beta
  • actin, cytoplasmic 1
  • B-Actin
  • Beta Actin
  • beta cytoskeletal actin
  • betaActin
  • beta-Actin
  • BRWS1
  • I(2)-Actin
  • PS1TP5-binding protein 1
  • PS1TP5BP1

Background

Actin is a highly conserved protein involved in cell motility, structure, and integrity. Six functional actin isoforms exist and were named based on their electrophoretic mobility and distribution including alpha-cardiac muscle actin (aCAA), alpha-skeletal muscle actin (aSKA), alpha-smooth muscle actin (aSMA), beta-cytoplasmic actin (bCYA), gamma-cytoplasmic actin (gCYA) and gamma-smooth muscle actin (gSMA) (1). The cytoplasmic isoforms, beta-actin and gamma-actin, are nearly identical, differing by only 4 amino acids at the N-terminus with a theoretical molecular weight of 42 kDa (2). Beta-actin is ubiquitously expressed in eukaryotic cells and considered a housekeeping gene which is frequently used as a loading control for assays involving protein detection, such as Western blotting (3,4).

References

1. Vandekerckhove J, Weber K. 1978. At least six different actins are expressed in a higher mammal: an analysis based on the amino acid sequence of the amino-terminal tryptic peptide. J Mol Biol. 126(4):783-802. PMID: 745245

2. Gimona M, Vandekerckhove J, Goethals M, Herzog M, Lando Z, Small JV. (1994) Beta-actin specific monoclonal antibody. Cell Motil Cytoskeleton. 27(2):108-16. PMID: 8162619

3. Holden VI, Lenio S, Kuick R, Ramakrishnan SK, Shah YM, Bachman MA. (2014) Bacterial siderophores that evade or overwhelm Lipocalin 2 induce HIF-1a and pro-inflammatory cytokine secretion in cultured respiratory epithelial cells Infect Immun. 82(9):3826-36 PMID: 24980968

4. Tsai WL, Yeh PH, Tsai CY, Ting CT, Chiu YH, Tao MH, Li WC, Hung SC. (2016) Efficient Programming of Human Mesenchymal Stem Cell Derived Hepatocytes by Epigenetic Regulations. J. Gastroenterol. Hepatol. 32(1):261-269. PMID: 27218433

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Publications for beta-Actin Antibody (NB600-503)(107)

We have publications tested in 11 confirmed species: Human, Mouse, Rat, Avian , Chinese Hamster, Fish, Human , Mouse , Porcine, Primate, Rabbit.

We have publications tested in 5 applications: B/N, IHC, IHC-P, PLA, WB.


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B/N
(1)
IHC
(1)
IHC-P
(2)
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(1)
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(54)
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Human
(27)
Mouse
(22)
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(13)
Avian
(1)
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(1)
Fish
(1)
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(2)
Mouse
(1)
Porcine
(2)
Primate
(1)
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(1)
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Showing Publications 1 - 10 of 107. Show All 107 Publications.
Publications using NB600-503 Applications Species
Hoover ME, Martin EC, Llamas CB et al. Proteomic characterization of a trauma-based rat model of heterotopic ossification identifies interactive signaling networks as potential therapeutic targets J Proteomics Jul 21 2020 [PMID: 32707234] (WB, Rat) WB Rat
Suter TACS, Blagburn SV, Fisher SE et al. TAG-1 Multifunctionality Coordinates Neuronal Migration, Axon Guidance, and Fasciculation Cell Rep Jan 28 2020 [PMID: 31995756] (WB, Mouse) WB Mouse
Fine JM, Kosyakovsky J, Baillargeon AM et al. Intranasal deferoxamine can improve memory in healthy C57 mice, suggesting a partially non-disease-specific pathway of functional neurologic improvement Brain Behav Jan 20 2020 [PMID: 31960628] (WB, Mouse) WB Mouse
Shastri S, Shinde T, Sohal SS et al. Idebenone Protects against Acute Murine Colitis via Antioxidant and Anti-Inflammatory Mechanisms Int J Mol Sci Jan 12 2020 [PMID: 31940911] (WB, Mouse) WB Mouse
Alhusaini A, Fadda L, Hasan IH et al. Arctium lappa Root Extract Prevents Lead-Induced Liver Injury by Attenuating Oxidative Stress and Inflammation, and Activating Akt/GSK-3beta Signaling Antioxidants (Basel) Nov 24 2019 [PMID: 31771282] (WB, Rat) WB Rat
Li J, Jeong SY, Xiong B et al. Repurposing pyridoxamine for therapeutic intervention of intravascular cell-cell interactions in mouse models of sickle cell disease Haematologica Oct 31 2019 [PMID: 31672902] (WB, Mouse, Human) WB Mouse, Human
Akhter ET, Griffith RW, English AW, Alvarez FJ Removal of the Potassium Chloride Co-Transporter from the Somatodendritic Membrane of Axotomized Motoneurons Is Independent of BDNF/TrkB Signaling But Is Controlled by Neuromuscular Innervation eNeuro Oct 16 2019 [PMID: 31541001] (IHC, Mouse) IHC Mouse
So EY, Sun C, Wu KQ et al. Lipid phosphatase SHIP-1 regulates chondrocyte hypertrophy and skeletal development J. Cell. Physiol. Jul 9 2019 [PMID: 31287165] (WB, Mouse) WB Mouse
Shah BS, Shimell JJ, Bamji SX Regulation of dendrite morphology and excitatory synapse formation by zDHHC15 Mol. Immunol. Jun 12 2019 [PMID: 31189538] (WB, Rat) WB Rat
Schneider JS, Aras R, Williams CK, et al. GM1 Ganglioside Modifies alpha-Synuclein Toxicity and is Neuroprotective in a Rat alplha-Synuclein Model of Parkinson's Disease Sci Rep 2019 Jun 10 [PMID: 31182727] (WB, Rat) WB Rat
Show All 107 Publications.

Reviews for beta-Actin Antibody (NB600-503) (4) 54

Average Rating: 5
(Based on 4 reviews)
We have 4 reviews tested in 3 species: Human, Mouse, Pig.

Reviews using NB600-503:
Filter by Applications
ICC
(2)
WB
(2)
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Human
(2)
Mouse
(1)
Pig
(1)
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Images Ratings Applications Species Date Details
Immunocytochemistry beta-Actin NB600-503
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reviewed by:
Anonymous
ICC Pig 12/05/2016
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Summary

ApplicationImmunocytochemistry
Sample Testedpig trabecular meshwork cells
SpeciesPig
Immunocytochemistry beta-Actin NB600-503
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reviewed by:
Anony- mous
ICC Human 06/11/2014
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Summary

ApplicationImmunocytochemistry
SpeciesHuman
Special ApplicationsIF Confocal analysis of HeLa cells using Beta Actin antibody (NB600-503, 1:5). An Alexa Fluor 488-conjugated Goat to rabbit IgG was used as secondary antibody (green. DAPI was used to stain the cell nuclei (blue).
FileView PDF
Western Blot beta-Actin NB600-503
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reviewed by:
Anony- mous
WB Human 06/11/2014
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Summary

ApplicationWestern Blot
SpeciesHuman
Special ApplicationsWestern blot analysis of extracts from Jurkat, COS and CHO cells using Actin antibody (NB600-503, 1:200).
FileView PDF
Western Blot beta-Actin NB600-503
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reviewed by:
Astrid Spannhoff
WB Mouse 11/19/2010
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Summary

ApplicationWestern Blot
Sample TestedNIH 3T3 cell lysate, Sample Amount: 5 ul
SpeciesMouse

Blocking

Blocking DetailsBlocking Buffer: 5% Milk in PBST, Blocking Time: 1 hour, Blocking Temp: 20 degrees Celsius

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1:5000, Incubation Dilution Buffer: 5% Milk in PBST, Incubation Time: overnight, Temp: 4C

Secondary Antibody

Secondary DescriptionSecondary Ab: anti-mouse IgG HRP labeled, Secondary Ab Dilution Ratio: 1:5000
Secondary Manufacturer Cat#NA9310

Details

Detection NotesDetection Method: ECL, Exposure Time: 30 seconds, Specific Bands: around 40 kDa

Product General Protocols

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for beta-Actin Antibody (NB600-503). (Showing 1 - 6 of 6 FAQs).

  1. Why is beta actin used as a loading control in blotting ?
    • Beta actin is a highly-expressed protein found in all cells, and is considered a house keeping protein whose expression is necessary for any cell's proper functioning. For this reason it is used as a loading control to confirm that the same amount of total protein is loaded in each lane of an SDS-PAGE gel so that appropriate comparisons can be made between the actual proteins of interest in different samples.
  2. Are the beta-Actin antibodies validated in Simple Western?
    • Yes, we offer a few beta actin antibodies that have been tested in Simple Western: NBP1-47423, NB600-501, NB600-532, NB600-503, and NB100-56874.
  3. Do Beta-Actin antibodies come in lyophilized format?
    • ACTB antibodies like MAB8969 AND MAB8929 come in lyophilized form.
  4. What is the theoretical molecular weight for Beta-Actin antibodies?
    • The TMW for beta Actin antibodies is approximately 42 kDa.
  5. I wanted to know which of the two housekeeping genes B-actin or GAPDH can be used for best results. I intend to do an experiment to determine expression of IL-6 and TNF-alpha in liver and kidney tissues.
    • For homogenized tissue, beta-actin and GAPDH are both fine.
  6. What is the immunogen sequence of this Beta-Actin antibody?
    • A sequence within the N-terminal region of Human beta Actin. The exact sequence is proprietary.

Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 NB600-503AF405
Alexa Fluor 488 NB600-503AF488
Alexa Fluor 647 NB600-503AF647
Alexa Fluor 700 NB600-503AF700
Allophycocyanin NB600-503APC
Biotin NB600-503B
DyLight 350 NB600-503UV
DyLight 405 NB600-503V
DyLight 488 NB600-503G
DyLight 550 NB600-503R
DyLight 594 NB600-503DL594
DyLight 650 NB600-503C
DyLight 680 NB600-503FR
DyLight 755 NB600-503IR
FITC NB600-503F
HRP NB600-503H
Janelia Fluor 549 NB600-503JF549
Janelia Fluor 646 NB600-503JF646
PE NB600-503PE
PerCP NB600-503PCP

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Blogs on beta-Actin.


  Read full blog post.

The use of Beta Actin (AC-15) as a loading control across multiple species
Actin is a fundamental component of the cytoskeleton, where it has the ability to create and break down actin filament formation in response to various cell needs.  Actin has six highly conserved isoforms, however beta and gamma actin are the two...  Read full blog post.

Tips on choosing an ideal loading control antibody for Western Blotting
Western blotting is one of the most commonly used antibody assay techniques in cell and molecular biology research since its development over three decades ago, and is considered the gold standard for protein detection and quantification. When p...  Read full blog post.

Beta-Actin's Role in Neuronal Plasticity
Beta-Actin is a highly conserved protein involved in cell growth, cytoskeletal and extracellular support structures and cell migration. Because beta-Actin is ubiquitously expressed in all eukaryotic cells, it is frequently used as a loading contro...  Read full blog post.

ACTB - an abundant cytoskeletal component with applications for gene expression analysis
Actin is the widely studied and ubiquitous cytoskeletal protein capable of forming dynamic microfilament structures. These filaments are essential for diverse cellular functions including cell shape, migration, cytokinesis, and intracellular traffi...  Read full blog post.

A New Standard in Antibody Testing - Simple Western Certified Antibodies
The Western blot is one of the most commonly used antibody assay techniques in cell and molecular biology research since its development over three decades ago, and is considered the gold standard for protein detection and quantification. The tradi...  Read full blog post.

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Recent Reviews

5
5
4
4
0
3
0
2
0
1
0

Anonymous
12/05/2016
Application: ICC
Species: Pig

Anony- mous
06/11/2014
Application: ICC
Species: Human

Anony- mous
06/11/2014
Application: WB
Species: Human

Bioinformatics

Gene Symbol ACTB
Entrez
Uniprot