MA5-15452

antibody from Invitrogen Antibodies

Targeting: ACTB

Western blot (Supportive data in Antibodypedia)

ELISA (Recommended by provider)

Immunocytochemistry (Supportive data in Antibodypedia)

Flow cytometry (Supportive data in Antibodypedia)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

MA5-15452

Provider

Invitrogen Antibodies

Product name

beta Actin Monoclonal Antibody (8H10D10)

Antibody type

Monoclonal

Antigen

Synthetic peptide

Description

MA5-15452 targets beta-Actin in indirect ELISA, FACS, IF, and WB applications and shows reactivity with Hamster, Human, mouse, Non-human primate, and Rat samples. The MA5-15452 immunogen is synthetic peptide corresponding to amino-terminal residues of human beta-Actin, conjugated to KLH. MA5-15452 detects beta-Actin which has a predicted molecular weight of approximately 42kDa.

Reactivity

Human, Mouse, Rat, Hamster

Host

Mouse

Isotype

IgG

Antibody clone number

8H10D10

Vial size

100 µL

Concentration

Conc. Not Determined

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Parecoxib inhibits esophageal squamous cell carcinoma progression via the PDK1-AKT pathway.

Huang HM, Huang XY, Wu SP, Chen CK, He XH, Zhang YF
Cellular & molecular biology letters 2022 Mar 19;27(1):28

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Low-Dose Albendazole Inhibits Epithelial-Mesenchymal Transition of Melanoma Cells by Enhancing Phosphorylated GSK-3β/Tyr216 Accumulation.

He Z, Lei S, Liang F, Tan L, Zhang W, Xie L, Zheng H, Lu Y
Journal of oncology 2021;2021:4475192

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The hepcidin regulator erythroferrone is a new member of the erythropoiesis-iron-bone circuitry.

Castro-Mollo M, Gera S, Ruiz-Martinez M, Feola M, Gumerova A, Planoutene M, Clementelli C, Sangkhae V, Casu C, Kim SM, Ostland V, Han H, Nemeth E, Fleming R, Rivella S, Lizneva D, Yuen T, Zaidi M, Ginzburg Y
eLife 2021 May 18;10

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Immunomodulatory effects of Radix isatidis polysaccharides in vitro and in vivo.

Tao W, Fu T, He ZJ, Zhou HP, Hong Y
Experimental and therapeutic medicine 2021 Dec;22(6):1405

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Cathepsin S Regulates Antigen Processing and T Cell Activity in Non-Hodgkin Lymphoma.

Dheilly E, Battistello E, Katanayeva N, Sungalee S, Michaux J, Duns G, Wehrle S, Sordet-Dessimoz J, Mina M, Racle J, Farinha P, Coukos G, Gfeller D, Mottok A, Kridel R, Correia BE, Steidl C, Bassani-Sternberg M, Ciriello G, Zoete V, Oricchio E
Cancer cell 2020 May 11;37(5):674-689.e12

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Prohibitin is a prognostic marker and therapeutic target to block chemotherapy resistance in Wilms' tumor.

Ortiz MV, Ahmed S, Burns M, Henssen AG, Hollmann TJ, MacArthur I, Gunasekera S, Gaewsky L, Bradwin G, Ryan J, Letai A, He Y, Naranjo A, Chi YY, LaQuaglia M, Heaton T, Cifani P, Dome JS, Gadd S, Perlman E, Mullen E, Steen H, Kentsis A
JCI insight 2019 Aug 8;4(15)

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Chromatin Remodeling in Response to BRCA2-Crisis.

Gruber JJ, Chen J, Geller B, Jäger N, Lipchik AM, Wang G, Kurian AW, Ford JM, Snyder MP
Cell reports 2019 Aug 20;28(8):2182-2193.e6

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Influence of Matrices on 3D-Cultured Prostate Cancer Cells' Drug Response and Expression of Drug-Action Associated Proteins.

Edmondson R, Adcock AF, Yang L
PloS one 2016;11(6):e0158116

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An Antibody-Drug Conjugate Directed against Lymphocyte Antigen 6 Complex, Locus E (LY6E) Provides Robust Tumor Killing in a Wide Range of Solid Tumor Malignancies.

Asundi J, Crocker L, Tremayne J, Chang P, Sakanaka C, Tanguay J, Spencer S, Chalasani S, Luis E, Gascoigne K, Desai R, Raja R, Friedman BA, Haverty PM, Polakis P, Firestein R
Clinical cancer research : an official journal of the American Association for Cancer Research 2015 Jul 15;21(14):3252-62

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of beta-Actin using beta-Actin monoclonal antibody (Product # MA5-15452) in NIH/3T3 (1), Jurkat (2), HeLa (3), CHO (4), PC12 (5), HEK293 (6), COS-7 (7), A549 (8) and MCF-7 (9) cell lysate.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of beta-Actin using beta-Actin monoclonal antibody (Product # MA5-15452) in NIH/3T3 (1), Jurkat (2), HeLa (3), CHO (4), PC12 (5), HEK293 (6), COS-7 (7), A549 (8) and MCF-7 (9) cell lysate.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis of beta-Actin using beta-Actin monoclonal antibody (Product # MA5-15452) in NIH/3T3 (1), Jurkat (2), HeLa (3), CHO (4), PC12 (5), HEK293 (6), COS-7 (7), A549 (8) and MCF-7 (9) cell lysate.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western Blot was performed using Anti-beta Actin Monoclonal Antibody (8H10D10) (Product # MA5-15452) and a ~40 kDa band corresponding to Actin, cytoplasmic 1 was observed across cell lines and tissues tested. Whole cell extracts (30 µg lysate) of HeLa (Lane 1), A549 (Lane 2), A-431 (Lane 3), U-2 OS (Lane 4), Jurkat (Lane 5), PC-12 (Lane 6), NIH/3T3 (Lane 7), Mouse Spleen (Lane 8), Mouse Liver (Lane 9), Rat Spleen (Lane 10), Rat Liver (Lane 11) were electrophoresed using NuPAGE™ 4-12% Bis-Tris Protein Gel (Product # NP0321BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # LC2001) by iBlot® 2 Dry Blotting System (Product # IB21001). The Blot was probed with the primary antibody (1:2000 dilution) and detected by chemiluminescence with Goat anti-Mouse IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A28177, 1:4000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005). doi:10.1038/bcj.2014.18

Immunocytochemistry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of SKBR-3 (left) and A549 (right) cells using beta-Actin monoclonal antibody (Product # MA5-15452) (Red). Blue: DRAQ5 fluorescent DNA dye. Red: the secondary Ab is Cy3-Goat anti mouse IgG

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of SKBR-3 (left) and A549 (right) cells using beta-Actin monoclonal antibody (Product # MA5-15452) (Red). Blue: DRAQ5 fluorescent DNA dye. Red: the secondary Ab is Cy3-Goat anti mouse IgG

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Immunofluorescence analysis of Actin, cytoplasmic 1 was performed using 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 5 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with beta Actin Monoclonal Antibody (8H10D10) (Product # MA5-15452) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Mouse IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 647 (Product # A32787), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing Cytoskeleton localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60x magnification.

Flow cytometry

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Flow cytometric analysis of MCF-7 cells using beta-Actin monoclonal antibody (Product # MA5-15452) (right) and negative control (left).

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

NULL

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 4 ABZ treatment downregulates the snail expression in melanoma cells by increasing the accumulation of phosphorylated GSK-3 beta /Tyr216. (a) The relative transcription levels of Snail in the ABZ-treated (0.4 mu M) and control groups of A375 (left) and B16-F10 (right) melanoma cells were measured by RT-qPCR, with beta -actin as the internal control. (b) The expression of transcription factor Snail in A375 (left) and B16-F10 (right) cells was detected by western blot analysis, with beta -actin as the internal reference protein. (c-d) The expression levels of cytoplasmic proteins AKT, pAKT, GSK-3 beta , pGSK-3 beta (Ser9/Tyr216) and Snail, and nuclear protein pSnail in A375 and B16-F10 cells were also determined by western blotting, with beta -actin and PCNA as the internal controls for the cytoplasmic and nuclear proteins, respectively. The histograms show the relative density of AKT/pAKT, GSK-3 beta /pGSK-3 beta (Ser9/Tyr216), and Snail/p-Snail. (e) A375 cells were cotreated with or without MG132 and 0.4 mu M ABZ for 24 h western blot (up) was used to detect the expression levels of AKT, pGSK-3 beta /Tyr216, Snail, N-cadherin, and E-cadherin in the cytoplasm of A375 cells. The histogram (bottom) shows the relative density of AKT, pGSK-3 beta /Tyr216, Snail, E-cadherin, and N-cadherin. (f) Histogram showing the relative expression intensity of pGSK-3 beta (Ser9/Tyr216) and pAKT after immunohistochemical staining of mouse metastatic lung cancer tissues. Scale bars = 100