Key Product Details

Species Reactivity

Validated:

Human, Mouse, Rat, Porcine, Bovine, Equine

Cited:

Bovine

Applications

Validated:

Flow Cytometry, Immunocytochemistry/ Immunofluorescence, Immunohistochemistry, Western Blot

Cited:

Block/Neutralize, In vivo assay, Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG1 Clone # 5J11

Concentration

1 mg/ml

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Product Specifications

Immunogen

An Actin preparation derived from bovine brain.

Localization

Cytoplasm > cytoskeleton.

Specificity

This antibody recognizes all six isotypes of Actin.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG1

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 pan Actin Antibody (5J11)

Western Blot: pan Actin Antibody (5J11) [NBP2-25142]

Western Blot: pan Actin Antibody (5J11) [NBP2-25142]

Western Blot: Actin Antibody (5J11) [NBP2-25142] - NBP2-25142 was used to detect GST fusion proteins expressing each of the six human actin proteins. GST-fusion proteins encoding all six full length human actin gene products were run out on SDS-PAGE, transferred to nitrocellulose and reacted with NBP2-25142. The antibody stained all six proteins indicating that it is a panspecific actin antibody.
Immunocytochemistry/ Immunofluorescence: pan Actin Antibody (5J11) [NBP2-25142]

Immunocytochemistry/ Immunofluorescence: pan Actin Antibody (5J11) [NBP2-25142]

Immunocytochemistry/Immunofluorescence: Actin Antibody (5J11) [NBP2-25142] - Beta actin was detected in NIH-3T3 cells fixed with methanol using mouse anti-mouse beta-Actin monoclonal antibody (NBP2-25142) at 1:2700 dilution. Cells were stained using Northern Lights 557 conjugated anti-mouse secondary antibody (NL007) and counterstained with DAPI.
Flow Cytometry: pan Actin Antibody (5J11) [NBP2-25142]

Flow Cytometry: pan Actin Antibody (5J11) [NBP2-25142]

Flow Cytometry: Actin Antibody (5J11) [NBP2-25142] - analysis of HeLa cells using mouse Monoclonal Actin antibody (Orange) and Isotype control Antibody (Blue).
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Applications for pan Actin Antibody (5J11)

Application
Recommended Usage

Flow Cytometry

1:10-1:1000

Immunocytochemistry/ Immunofluorescence

1:500-1:1000

Immunohistochemistry

1:500-1:1000

Western Blot

1:1000
Application Notes
This Actin (5J11) antibody is useful for Immunocytochemistry/Immunofluorescence, Immunohistochemistry, and Western Blot, where a band can be seen at ~42 kDa.

Reviewed Applications

Read 2 reviews rated 5 using NBP2-25142 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

50% PBS, 50% glycerol

Preservative

0.035% 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 short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: pan Actin

Actin is one of the most abundant and highly conserved proteins of eukaryotes. Mammalian Actins are the product of six different genes with differing distribution patterns in cell types and in tissues. The molecular weight of all six proteins is 42 kDa, and one or more actins are found in essentially every type of crude cellular and tissue extract. As a result, antibodies to Actin are widely used. As western blotting standards, they can be used to verify that the various steps of the western blotting procedure have been performed correctly. In addition, Actin is regarded as a "housekeeping" protein which is generally not altered much in expression as a result of experimental manipulations. Thus, quantitation of the Actin band on the western is used as a standard against the band density of other proteins. The Actin isotypes were originally classified as alpha, beta and gamma forms based on their charges which resulted in different mobilities in the isoelectric focusing dimension as seen on 2-dimensional SDS-PAGE. Subsequently, the alpha spot was found to potentially contain three Actin gene products, alpha-skeletal Actin, alpha-vascular smooth muscle Actin, and alpha-cardiac muscle. The beta spot contained a single protein called simply beta Actin, while the gamma spot may contain both gamma-1 Actin and gamma-2, which are enteric and smooth muscle Actins. The six mammalian gene products are very similar as shown by a sequence alignment of the human proteins. The most closely related actin isotypes are 97% identical, while the most distantly related are 94% identical, with most of the variability at the N-terminus. With this level of identity, antibodies to any one protein may recognize all six isotypes, as is the case with clone 5J11. This antibody binds all six isotypes very strongly on western blots, and it is a very effective blotting standard which can work on any cell type or tissue extract. It also works in immunocytochemical experiments, binding strongly and cleanly to filopodia, membrane ruffles and stress fibers, all known to be rich in Actin.

Alternate Names

ACTA, actin, alpha 1, skeletal muscle, alpha skeletal muscle, alpha skeletal muscle actin, Alpha-actin-1, ASMA, CFTD, CFTDM, MPFD, NEM1, NEM2, NEM3

Gene Symbol

ACTA1

Additional pan Actin Products

Product Documents for pan Actin Antibody (5J11)

Product Datasheets

Certificate of Analysis

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Product Specific Notices for pan Actin Antibody (5J11)

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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