Log In Sign up
The largest database of trusted experimental protocols

Smad3 9523

Manufactured by Cell Signaling Technology
Visit Supplier
Sourced in United States

Smad3 #9523 is a primary antibody that recognizes the Smad3 protein. Smad3 is a key mediator of the transforming growth factor-beta (TGF-β) signaling pathway. This antibody can be used to detect Smad3 expression in various cell and tissue samples.

Automatically generated - may contain errors

Lab products found in correlation

β actin, by Merck Group (3 mentions) P smad3 9520, by Cell Signaling Technology (3 mentions) Erk1 2 9102, by Cell Signaling Technology (2 mentions) Akt 4691, by Cell Signaling Technology (2 mentions) P smad2 3108, by Cell Signaling Technology (2 mentions) Smad2 3103, by Cell Signaling Technology (2 mentions) P akt 4058, by Cell Signaling Technology (2 mentions) Perk1 2 9101, by Cell Signaling Technology (2 mentions) Anti mouse 5470 and anti rabbit 5151 secondary antibodies, by Cell Signaling Technology (2 mentions) P38 9212, by Cell Signaling Technology (2 mentions) Slug 9585, by Cell Signaling Technology (2 mentions) N cadherin, by BD (2 mentions) E cadherin, by BD (2 mentions) Chemiluminescent peroxidase substrate, by Merck Group (1 mentions) Pvdf nitrocellulose membrane, by Merck Group (1 mentions) Gapdh ap0063, by Bioworld Technology (1 mentions) Prdm16 ab106410, by Abcam (1 mentions) Bca protein assay kit, by Beyotime (1 mentions) Chemidoc xrs system, by Bio-Rad (1 mentions) Fluo 4, by Thermo Fisher Scientific (1 mentions)

Spelling variants of this product

Smad3 (242 citations) Anti-SMAD3 (#9523; (3 citations)

7 protocols using smad3 9523

1

Browne Adipose Tissue Protein Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protein from brown adipocytes or brown adipose tissue was extracted using lysing buffer. Protein concentration was determined using BCA Protein Assay kit (Beyotime Institute of Biotechnology, Nanjing, China). Proteins (30 μg) were separated by SDS-PAGE, transferred to PVDF nitrocellulose membrane (Millipore, Boston, MA, USA), blocked with 5% fat-free milk for 2 h at room temperature and then incubated with primary antibodies in 5% milk overnight at 4°C. Sirt1 (ab110304), Sirt2 (ab191383), CHOP (ab179823), GRP78 (ab108615), ATF4 (ab184909), ERDJ4 (ab118282), Bax (ab32503), Apaf-1 (ab32372), UCP1 (ab2384) and PRDM16 (ab106410) antibodies were all purchased from Abcam (Cambridge, UK). Active-Caspase 3 (bs7004), active-Caspase 9 (bs7070), Bcl-2 (bs1511) and GAPDH (ap0063) antibodies were purchased from Bioworld (Nanjing, China). Smad3 (9523S) and IRE1 (3294S) antibodies were purchased from Cell Signaling Technology (CST, Boston, MA, USA). Rabbit HRP-conjugated secondary antibody (Boaoshen, Beijing, China) was added and incubated at room temperature for 2 h. Proteins were visualized using chemiluminescent peroxidase substrate (Millipore, Boston, MA, USA), and then the blots were quantified using ChemiDoc XRS system (Bio-Rad, Hercules, CA, USA).
Liu Z., Gu H., Gan L., Xu Y., Feng F., Saeed M, & Sun C. (2016). Reducing Smad3/ATF4 was essential for Sirt1 inhibiting ER stress-induced apoptosis in mice brown adipose tissue. Oncotarget, 8(6), 9267-9279.
+ Open protocol
+ Expand
2

Cellular Signaling Pathway Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols
Recombinant activin A and FST were purchased from R&D systems (Minneapolis, Mn, United States). DNase I and Liberase™ Research Grade were obtained from Roche (Mannheim, Germany). Progesterone (P4) and Estradiol (E2) were provided by TCI (Shanghai, China). Giemsa cytological stain was purchased from Sigma Aldrich (Oakville, ON, Canada). Fluo‐4 was from Thermo Fisher Scientific (Ottawa, ON, Canada). The antibodies against GAPDH (abs132004), MMP2 (abs130432) and MMP9 (abs155182) were obtained from Absin (Shanghai, China). The antibodies against Ezrin (3145S), Vimentin (5741S), p‐ERK1/2 (4370S), ERK1/2 (4695S), p‐SMAD3 (9520S) and SMAD3 (9523S) were purchased from Cell Signaling Technology (Danvers, MA, United States). The antibodies against p‐p38 (WLP1576), p38 (WL00764), p‐JNK (WL01813) and JNK (WL01295) were bought from Wanlei Biotechnology (Shenyang, China). JNK inhibitor AS601245 was obtained from Absin (Shanghai, China).
Liu G., Qi Y., Wu J., Lin F., Liu Z, & Cui X. (2022). Follistatin is a crucial chemoattractant for mouse decidualized endometrial stromal cell migration by JNK signalling. Journal of Cellular and Molecular Medicine, 27(1), 127-140.
+ Open protocol
+ Expand
3

Western Blot Analysis of Smad Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols
Protein lysates were subjected to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and then transferred to a PVDF membrane. Membranes were blocked and incubated, according to the instructions provided, with the primary antibody. Protein bands were identified through a horseradish peroxidase-conjugated secondary antibody and enhanced chemiluminescence reagents (Pierce, Rockford, IL). The following primary antibodies were used for Western blots: phospho-Smad2 (Ser465/467) #3108, Smad2 #5339, phospho-Smad3 (Ser423/425) #9520, and Smad3 #9523 (each 1:1000, Cell Signaling Technology); CS-56 (1:500, C8035, Sigma-Aldrich); β-actin (1:3000, A5316, Sigma-Aldrich); GFAP (1:1000, Z0334, Dako); KCa3.1 (1:500, ab83740, Abcam). The following secondary antibodies were used: HRP-linked anti-rabbit IgG (1:2000, NA9340V, GE Healthcare); HRP-linked anti-mouse IgG (1:2000, NA9310V, GE Healthcare). Immunoreactivity for each protein was quantified using Image J software. Blots of cell lysate were reprobed with β-actin as a loading control.
Yu Z., Yu P., Chen H, & Geller H.M. (2014). Targeted inhibition of KCa3.1 attenuates TGF-β-induced reactive astrogliosis through the Smad2/3 signaling pathway. Journal of neurochemistry, 130(1), 41-49.
+ Open protocol
+ Expand
4

TGF-β Signaling Pathway Activation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were serum-starved overnight and treated with 100pM TGF-β for 60 minutes. Cells were lysed in 2× Laemmli sample buffer, and the proteins were resolved on a 10% SDS-PAGE gel. All primary antibodies (Slug #9585, p-Smad2 #3108, Smad2 #3103, p-Smad3 #9520, Smad3 #9523, p-ERK1/2 #9101, ERK1/2 #9102, p-AKT #4058, AKT #4691, p-p38 #4631, p38 #9212) were purchased from Cell Signaling (Danvers, MA, USA) except for β-actin (Sigma, AA5441) and N-cadherin (#610921) and E-cadherin (#610182) (BD Biosciences). Anti-mouse (#5470) and anti-rabbit (#5151) secondary antibodies were purchased from Cell Signaling. Blots were scanned using the LI-COR Odyssey (LI-COR Biosciences, Lincoln, NE, USA).
Huang J.J., Corona A.L., Dunn B.P., Cai E.M., Prakken J.N, & Blobe G.C. (2019). Increased type III TGF-β receptor shedding decreases tumorigenesis through induction of epithelial-to-mesenchymal transition. Oncogene, 38(18), 3402-3414.
+ Open protocol
+ Expand
5

Protein Expression Profiling in Cell Signaling

Check if the same lab product or an alternative is used in the 5 most similar protocols
Total protein was isolated using RIPA lysis buffer, separated by 10% sodium dodecyl sulfate polyacrylamide gel electrophoresis, and then transferred onto polyvinylidene fluoride membranes. The membrane was and probed with diluted primary rabbit antibodies against Caspase-3 (ab13847), Bcl-2-associated X protein (Bax; ab32503), B-cell lymphoma 2 (Bcl-2) (ab32124), Bcl-XL (ab32370), TGF-β1 (ab92486), β-actin (ab8227), GAPDH (ab181602), E-cadherin (3195), N-cadherin (13116), matrix metalloproteinase (MMP)3 (14351), MMP9 (13667), Smad2 (5339), Smad3 (9523), phosphorylated (p)-Smad2 (18338), and p-Smad3 (9520, Cell Signaling Technologies) overnight at 4°C. The antibodies were from Abcam except for p-Smad2/3. After washing, membrane was re-probed with the horseradish peroxidase-conjugated goat anti-rabbit secondary antibody (ab205719, 1: 2000, Abcam) for 1 h. The protein bands were visualized using enhanced chemiluminescence (EMD Millipore). β-actin and GAPDH were used as internal controls. The gray values were analyzed with Image J software.
Li Z., Li M., Xia P., Wang L, & Lu Z. (2022). Targeting long non-coding RNA PVT1/TGF-β/Smad by p53 prevents glioma progression. Cancer Biology & Therapy, 23(1), 225-233.
+ Open protocol
+ Expand
6

TGF-β Signaling Pathway Activation

Check if the same lab product or an alternative is used in the 5 most similar protocols
Cells were serum-starved overnight and treated with 100pM TGF-β for 60 minutes. Cells were lysed in 2× Laemmli sample buffer, and the proteins were resolved on a 10% SDS-PAGE gel. All primary antibodies (Slug #9585, p-Smad2 #3108, Smad2 #3103, p-Smad3 #9520, Smad3 #9523, p-ERK1/2 #9101, ERK1/2 #9102, p-AKT #4058, AKT #4691, p-p38 #4631, p38 #9212) were purchased from Cell Signaling (Danvers, MA, USA) except for β-actin (Sigma, AA5441) and N-cadherin (#610921) and E-cadherin (#610182) (BD Biosciences). Anti-mouse (#5470) and anti-rabbit (#5151) secondary antibodies were purchased from Cell Signaling. Blots were scanned using the LI-COR Odyssey (LI-COR Biosciences, Lincoln, NE, USA).
Huang J.J., Corona A.L., Dunn B.P., Cai E.M., Prakken J.N, & Blobe G.C. (2019). Increased type III TGF-β receptor shedding decreases tumorigenesis through induction of epithelial-to-mesenchymal transition. Oncogene, 38(18), 3402-3414.
+ Open protocol
+ Expand
7

Analyzing NF-κB and Smad Signaling in Ac-HSC

Check if the same lab product or an alternative is used in the 5 most similar protocols
We further examined the in vitro effects of INT747 and losartan on nuclear factor kappa B (NF‐κB) and mothers against decapentaplegic (Smad) signaling pathways in Ac‐HSC, using western blot analysis. A 50‐μg protein was resolved on 10% acrylamide sodium dodecyl sulfate–polyacrylamide gel electrophoresis gels and transferred to polyvinylidene fluoride membranes by semidry transfer. Membranes were blocked using 5% skim milk in Tris‐buffered saline with Tween 20 (TBST) for 1 hour at room temperature, followed by overnight incubation with 1/1,000 dilution of specific antibodies against phosphorylated (p) ‐NF‐κB (3031), NF‐κB (8242) (Bioss Inc., Woburn, MA), p‐Smad2 (3101), p‐Smad3 (9520), Smad2/3 (3102), and Smad3 (9523) (Cell Signaling Technology Inc., Beverly, MA) at 4 °C; the membranes were then washed in TBST before incubating with horseradish peroxidase‐conjugated secondary antibodies (diluted in 3% [weight per volume] bovine serum albumin in TBST; 1:1,000) for 1 hour at room temperature. After additional washing in TBST, signals were detected using enhanced chemiluminescence, and densitometric scans were quantified using Image J software (NIH).
Namisaki T., Moriya K., Kitade M., Takeda K., Kaji K., Okura Y., Shimozato N., Sato S., Nishimura N., Seki K., Kawaratani H., Takaya H., Sawada Y., Akahane T., Saikawa S., Nakanishi K., Kubo T., Furukawa M., Noguchi R., Asada K., Kitagawa K., Ozutsumi T., Tsuji Y., Kaya D., Fujinaga Y, & Yoshiji H. (2017). Effect of combined farnesoid X receptor agonist and angiotensin II type 1 receptor blocker on hepatic fibrosis. Hepatology Communications, 1(9), 928-945.
+ Open protocol
+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required

Sign up now

Revolutionizing how scientists
search and build protocols!