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Key Product Details

Species Reactivity

Human, Mouse

Applications

ICC/IF (Negative), Immunohistochemistry, Immunoprecipitation (Negative), Simple Western, Western Blot

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Concentration

1 mg/ml

Product Summary for beta-Actin Antibody

Immunogen

This beta-Actin Antibody maps to a region corresponding to the N-terminus of human Beta Actin. [UniProt# P60709]

Predicted Species

Bovine (100%), Canine (100%), Chicken (100%), Chimpanzee (100%), Golden Syrian Hamster (100%), Guinea Pig (100%), Orangutan (100%), Porcine (100%), Rabbit (100%), Rat (100%), Sheep (100%), Xenopus (100%). Backed by our 100% Guarantee.

Reactivity Notes

Based on 100% sequence identity, this antibody is predicted to react with Rat, X. tropicalis, Chicken, Sheep, Bovine, Dog, Horse, Rabbit, Guinea pig, Pig, Golden hamster, Orangutan, and Chimpanzee.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

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.

Scientific Data Images for beta-Actin Antibody

Simple Western: beta-Actin Antibody [NB600-532] - Simple Western lane view shows a specific band for Beta Actin using NB600-532 at 1:200 in A431, C2C12 and C6 cell lysates. This experiment was performed under reducing conditions using the 12-230 kDa separation system.
Western Blot: beta-Actin Antibody [NB600-532] - Detection of actin in 3T3 lysates (20ug). ECL detection 30 seconds. A specific band is seen using different dilutions: Lane 1 (1:15,000), Lane 2 (1:10,000), and Lane 3 (1:15,000).
Western Blot: beta-Actin Antibody [NB600-532] - Detection of human and mouse Cytoskeletal Actin by western blot. Samples: Whole cell lysate (50 ug) from HeLa, HEK293T, Jurkat, mouse TCMK-1, and mouse NIH 3T3 cells prepared using NETN lysis buffer. Antibody: Affinity purified rabbit anti-Cytoskeletal Actin antibody NB600-532 used for WB at 0.1 ug/ml. Detection: Chemiluminescence with an exposure time of 1 second.

Applications for beta-Actin Antibody

Application
Recommended Usage

Immunohistochemistry

1:2000-1:10000

Simple Western

1:2000

Western Blot

1:2000-1:10000
Application Notes
This antibody is useful for Western Blot. A 40 kDa band is detected in HeLa whole cell lysate and mouse NIH3T3 cells. For IHC, epitope retrieval with citrate buffer pH6.0 is recommended for FFPE tissue sections.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. Separated by Size-Wes, Sally Sue/Peggy Sue.
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 6 reviews rated 4.5 using NB600-532 in the following applications:

Published Applications

Read 64 publications using NB600-532 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

Tris-Citrate/Phosphate (pH 7.0 - 8.0)

Preservative

0.09% Sodium Azide

Concentration

1 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C. Do not freeze.

Background: beta-Actin

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

Alternate Names

ACTB, betaActin

Gene Symbol

ACTB

Product Documents for beta-Actin Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for beta-Actin Antibody

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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Protocols

View specific protocols for beta-Actin Antibody (NB600-532):

[[URL:https://www.novusbio.com/products/beta-actin-antibody_nb600-532]][[Capt… Antibody]]
Western Blot Protocol

1. Perform SDS-PAGE (4-12%) on samples to be analyzed, loading 20 ug of total protein per lane.

2. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.

3. Stain the blot using ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.

4. Rinse the blot in TBS for approximately 5 minutes.

5. Block the membrane using 5% non-fat dry milk + 1% BSA in TBS for 1 hour.

6. Dilute the rabbit anti-actin primary antibody (NB 600-532) in blocking buffer and incubate 2 hours at room temperature.

7. Wash the membrane in water for 5 minutes and apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) and incubate 1 hour at room temperature.

8. Wash the blot in TBS containing 0.05-0.1% Tween-20 for 10-20 minutes.

9. Wash the blot in type I water for an additional 10-20 minutes (this step can be repeated as required to reduce background).

10. Apply the detection reagent of choice in accordance with the manufacturer's instructions (Amersham's ECL is the standard reagent used at Novus Biologicals).

Note: Tween-20 can be added to the blocking buffer at a final concentration of 0.05-0.2%, provided it does not interfere
with antibody-antigen binding.

FAQs for beta-Actin Antibody

Showing  1 - 2 of 2 FAQs Showing All
  • Q: I would like to know whether the antibody anti-actin (NB600-532) from rabbit can be also used for immunofluorescence and which dilution you recommend?

    A: This antibody is listed as being negative for immunocytochemistry/immunofluorescence, so is not recommended for your assay. As it has been validated in ICC/IF, I would highly recommend NB600-501 instead of NB600-532. Full information about this product and recommended dilutions can be found on the datasheet. For ICC/IF, we recommend a dilution of 1:1000-1:2000.

  • Q: What is the immunogen sequence of this Beta-Actin antibody?

    A: A sequence within the N-terminal region of Human beta Actin. The exact sequence is proprietary.

Showing  1 - 2 of 2 FAQs Showing All
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