Catalog # PA1-183

beta Actin Polyclonal Antibody

Product Details
Size 100 µg
Species Reactivity Human, Mouse, Non-human primate, Rat
Published Species Rat, Mouse, Human
Host/Isotope Rabbit / IgG
Class Polyclonal
Type Antibody
Conjugate Unconjugated
Immunogen Purified full-length recombinant human beta-actin expressed in E. coli
Form Liquid
Concentration 1 mg/mL
Purification Protein G
Storage buffer PBS with 30% glycerol, 1mg/mL BSA
Contains 0.05% sodium azide
Storage Conditions -20° C, Avoid Freeze/Thaw Cycles
RRID AB_2539914

Applications Tested Dilution Publications

Immunocytochemistry (ICC) 1:50-1:200 -
Immunofluorescence (IF) 1:50 - 1:200 -
Immunoprecipitation (IP) 2 µg 1 Publication
Western Blot (WB) 1:500 - 1:2000 11 Publications
Miscellaneous PubMed (Misc) - 1 Publication

Product Specific Information

By Western blot, PA1-183 detects a prominent ~42kD protein in human, mouse, non-human primate, and rat whole cell lysates.

For visualization of actin filaments by immunofluorescence, fixation and permeabilization with methanol is required.

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 Product Images For beta Actin Polyclonal Antibody

beta Actin Antibody (PA1-183) in IF

Immunofluorescent analysis of beta-actin (green) in HeLa cells. Cells were fixed with
4% paraformaldehyde (PFA) for 15 minutes at room temperature and permeabilized
with 0.5% Triton X-100 in PBS for 15 minutes at room temperature (left panel) or
fixed and permeabilized with ice cold methanol for 10 minutes at room temperature
(middle panel). Cells were blocked with 0.3% BSA in PBS for 15 minutes at room
temperature, and probed with a beta-actin polyclonal antibody (Product # PA1-183)
at a dilution of 1:100 for at least 1 hour at room temperature. Cells were then washed
with PBS, and incubated with a DyLight 488 goat anti-rabbit IgG secondary antibody
(Product # 35552) at a dilution of 1:500 for 30 minutes at room temperature. As a
positive control, PFA-fixed cells were stained with DyLight 488-conjugated Phalloidin
(Product # 21833) at a dilution of 1:1000 (right panel). Nuclei (blue) were stained with
Hoechst 33342 dye (Product # 62249). Images were taken on a Thermo Scientific
ToxInsight Instrument at 20X magnification.

beta Actin Antibody (PA1-183) in WB

Western blot analysis of beta-actin was performed by loading 25 µg of various whole
cell lysates and 10 µL of PageRuler Plus Prestained Protein Ladder (Product #
26619) per well onto a 4-20% Tris-HCl polyacrylamide gel. Proteins were transferred
to a PVDF membrane and blocked with 5% BSA/TBST for at least 1 hour. The
membrane was probed with a beta-actin polyclonal antibody (Product # PA1-183) at
a dilution of 1:1000 overnight at 4°C on a rocking platform, washed in TBS-0.1%
Tween-20, and probed with a goat anti-rabbit IgG-HRP secondary antibody (Product
# 31460) at a dilution of 1:40,000 for at least 1 hour. Chemiluminescent detection
was performed using SuperSignal West Pico (Product # 34080).

beta Actin Antibody (PA1-183) in IP

Immunoprecipitation of beta-actin was performed on HeLa cells. Antigen-antibody
complexes were formed by incubating 750 µg of HeLa whole cell lysates with 2 µg of
a beta-actin polyclonal antibody (Product # PA1-183) overnight on a rocking platform
at 4°C. The immune complexes were captured on 50 µL Protein A/G Agarose
(Product # 20421), washed extensively, and eluted with 5X Lane Marker Reducing
Sample Buffer (Product # 39000). Samples were resolved on a 4-20% Tris-HCl
polyacrylamide gel, transferred to a PVDF membrane, and blocked with 5% BSA
/TBS-0.1%Tween-20 for at least 1 hour. The membrane was probed with a beta-
actin polyclonal antibody (Product # PA1-183) at a dilution of 1:1000 overnight
rotating at 4°C, washed in TBST, and probed with Clean-blot IP detection reagent
(Product # 21230) at a dilution of 1:250 for at least 1 hour. Chemiluminescent
detection was performed using SuperSignal West Pico (Product # 34080).

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 13 References

Western Blot (11)

Frontiers in behavioral neuroscience Species

Rat
Fenofibrate Administration Reduces Alcohol and Saccharin Intake in
Rats: Possible Effects at Peripheral and Central Levels. Dilution
Not Cited
"PA1-183 was used in Western Blotting to identify that reduction of ethanol intake by fenofibrate is a consequence of
catalase induction in the liver and central pharmacological effects."
Year
Authors: Rivera-Meza M,Muñoz D,Jerez E,Quintanilla ME,Salinas-Luypaert C,Fernandez K,Karahanian E 2020

Aging cell Species

Mouse
Systemic clearance of p16INK4a -positive senescent cells mitigates age-
associated intervertebral disc degeneration. Dilution
Not Cited
"PA1-183 was used in Western Blotting to determine the relationship between cellular senescence and and age-related
intervertebral degeneration."
Year
Authors: Patil P,Dong Q,Wang D,Chang J,Wiley C,Demaria M,Lee J,Kang J,Niedernhofer LJ,Robbins PD,Sowa G, 2019
Campisi J,Zhou D,Vo N

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Immunoprecipitation (1)

Experimental and therapeutic medicine Species

Human
1,25-(OH)2D3 and its analogue BXL-628 inhibit high glucose-induced
Dilution
activation of RhoA/ROCK pathway in HK-2 cells. 1:5,000
"PA1-183 was used in Immunoprecipitation to demonstrate that 1,25-(OH)2D3 and its analog BXenopus laevis-628
contribute to renoprotection by preventing high glucose-induced EMT and ECM accumulation via inactivating the RhoA Year
/ROCK signaling pathway in vitro." 2017
Authors: Zhang W,Yi B,Zhang K,Li A,Yang S,Huang J,Liu J,Zhang H

Miscellaneous PubMed (1)

Experimental and therapeutic medicine Species

Human
Overexpression of the pituitary tumor transforming gene upregulates
metastasis in malignant neoplasms of the human salivary glands. Dilution
1:300
"PA1-183 was used in immunohistochemistry - paraffin section and western blot to examine the contribution of the
pituitary tumor transforming gene to salivary gland malignant neoplasms"
Year
Authors: Liu J,Wang Y,He H,Jin W,Zheng R 2015

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For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization. Products are warranted to operate or perform substantially in conformance with published Product specifications in effect at the time of sale, as set forth in the Production documentation,
specifications and/or accompanying package inserts ("Documentation"). No claim of suitability for use in applications regulated by FDA is made. The warranty provided herein is valid only when used by properly trained individuals. Unless otherwise stated in the Documentation, this warranty is limited to
one year from date of shipment when the Product is subjected to normal, proper and intended usage. This warranty does not extend to anyone other than the Buyer. Any model or sample furnished to Buyer is merely illustrative of the general type and quality of goods and does not represent that any
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expressly stated on the Product or in the documentation accompanying the Product, the Product is intended for research only and is not to be used for any other purpose, including without limitation, unauthorized commercial uses, in vitro diagnostic uses, ex vivo or in vivo therapeutic uses, or any type
of consumption by or application to human or animals.

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