Gata4

Manufactured by Cell Signaling Technology

Sourced in United States

Gata4 is a transcription factor that plays a crucial role in the regulation of gene expression. It is involved in the development and function of various tissues, particularly the cardiovascular system and the gastrointestinal tract.

Automatically generated - may contain errors

Lab products found in correlation

Spelling variants of this product

Anti-GATA4 (4 citations)

11 protocols using gata4

Protein Analysis via SDS-PAGE and Western Blotting

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed, and protein levels were determined by SDS-PAGE coupled to Western blotting. Additional details for this protocol are provided in the Supplementary Materials. Antibodies were obtained commercially and included GATA4 (1:1000, Cell Signaling Technology, Danvers, MA, USA) and COL1α1 (1:1000, Aviva Systems Biology, San Diego, CA, USA). The secondary antibody was anti-rabbit (1:10,000, Santa Cruz Biotechnology Inc., Dallas, TX, USA). Image analysis was performed using Image Lab 6.0 software from Bio-Rad (Berkeley, CA, USA).

Gallego P., Luque-Sierra A., Falcon G., Carbonero P., Grande L., Bautista J.D., Martín F, & Del Campo J.A. (2021). White Button Mushroom Extracts Modulate Hepatic Fibrosis Progression, Inflammation, and Oxidative Stress In Vitro and in LDLR-/- Mice. Foods, 10(8), 1788.

+ Open protocol

+ Expand

Protein Expression Analysis Using Western Blot

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were lysed in buffer containing RIPA and PSMF (TransGen Biotech, China). Proteins were quantified using a BCA protein assay kit (Beyotime Biotechnology) according to the manufacturer’s specifications. After denaturation, proteins were separated on 10% polyacrylamide gels and then transfered to polyvinylidene fluoride (PVDF) membranes (Millipore, CA, USA). After being blocked, the membranes were incubated overnight with primary antibodies for E-cadherin, Vimentin, Gata4, Sox2, Foxa2, Desmin (Cell Signalling Technology), Sox17 (R&D Systems), and Gadph (ProteinTech) at 4 °C and then with secondary antibodies on the next day. Finally, the protein bands were visualised with hypersensitive chemiluminescence (Beyotime Biotechnology).

He X., Chi G., Li M., Xu J., Zhang L., Song Y., Wang L, & Li Y. (2020). Characterisation of extraembryonic endoderm-like cells from mouse embryonic fibroblasts induced using chemicals alone. Stem Cell Research & Therapy, 11, 157.

+ Open protocol

+ Expand

Western Blot Antibody Catalog

Check if the same lab product or an alternative is used in the 5 most similar protocols

PDE3A (catalog number A302-740A, 1:1000 [Bethyl Laboratory]; catalog number Sc-293446, 1:20 [Santa Cruz Biotechnology]), Flag (catalog number F7425, 1:2000; Sigma), GAPDH (catalog number 60004-1-1g, 1:5000; ProteinTech), Anti-phospho-PKA substrate antibody (catalog number 9621S or 9624, 1:1000; Cell Signaling), SMAD4 (catalog number 46535, GTX01674; Cell Signaling) GATA4 (GATA4 binding protein 4; catalog number 36966, 1:1000; Cell Signaling), HDAC-1 (catalog number 34589, 1:1000; Cell Signaling), lamin B (catalog number 12586, 1:1000; Cell Signaling), lamin A/C (catalog number 4777S, 1:1000; Cell Signaling), actin-1 alpha (catalog number 17521-1-AP; Proteintech), RFP (catalog number ab62341, 1:2000; Abcam), GFP (catalog number ab6556, 1:1000; Abcam), horseradish peroxidase–conjugated goat anti-mouse (catalog number sc2005, 1:5000 [SCBT]; catalog number 705-035-151 [Jackson ImmunoResearch Labs]) or anti-rabbit secondary antibodies (catalog number sc2030, 1:5000 [Santa Cruz Biotechnology]; catalog number 705-035-152 [Jackson ImmunoResearch Labs]), antibodies conjugated with Alexa-Fluor 647 (catalog number A-21235, 1:250; Thermo Fisher Scientific), Alexa-Fluor 488 (catalog number A-11008, 1:250; Thermo Fisher Scientific), and wheat germ agglutinin conjugated with Alexa 488 (catalog number W11261; Invitrogen).

Subramaniam G., Schleicher K., Kovanich D., Zerio A., Folkmanaite M., Chao Y.C., Surdo N.C., Koschinski A., Hu J., Scholten A., Heck A.J., Ercu M., Sholokh A., Park K.C., Klussmann E., Meraviglia V., Bellin M., Zanivan S., Hester S., Mohammed S, & Zaccolo M. (2023). Integrated Proteomics Unveils Nuclear PDE3A2 as a Regulator of Cardiac Myocyte Hypertrophy. Circulation Research, 132(7), 828-848.

+ Open protocol

+ Expand

Western Blot Analysis of Cardiac Markers

Check if the same lab product or an alternative is used in the 5 most similar protocols

Cells were homogenized in ice cold lysis buffer containing proteinase and phosphatase inhibitor cocktail. The primary antibodies utilized in this study were as follows: OCT4 (sc-5279; Santa Cruz Biotechnology), Nanog (A3233 ABclonal), NKX2.5 (ab91196; Abcam), α-actinin (6487; Cell Signaling Technology), CX43 (3512 Cell Signaling Technology), GATA4 (5851; Cell Signaling Technology), MEF2C (5030; Cell Signaling Technology), cTnI (ab47003; Abcam), p-ERK (4370; Cell Signaling Technology), ERK (5013 Cell Signaling Technology), β-Catenin (8480; Cell Signaling Technology), p-cadherin (2189; Cell Signaling Technology), p-STAT3 (RT1490; HuaBio), STAT3 (ET1605; HuaBio), and GAPDH (5174; Cell Signaling Technology). Blots were developed using an enhanced chemiluminescence reagent (Thermo Fisher Scientific, Waltham, MA, USA).

Zhu X., Ding S., Li H., Zhang Z., Xu L., Wu J., Wang X., Zou Y., Yang X, & Ge J. (2020). Disruption of histamine/H1R signaling pathway represses cardiac differentiation and maturation of human induced pluripotent stem cells. Stem Cell Research & Therapy, 11, 27.

+ Open protocol

+ Expand

Western Blot Analysis of HCMV Infection

Check if the same lab product or an alternative is used in the 5 most similar protocols

Lysates of uninfected or infected HFF cells (see text and figure legends) were prepared for Western blotting by washing cells once in PBS and then suspending them directly in 2× Laemmli buffer containing 5 % β-mercaptoethanol before incubation at 95 °C for 5 min. Western blotting of proteins separated on 8 or 10 % polyacrylamide gels was carried out as described elsewhere [24 (link)], using antibodies recognizing IE1/2, UL44, pp28, (all Virusys, 1 : 1000 dilution), β-actin (SIGMA, 1 : 5000 dilution), PDGFRa (ABCAM ab203491, 1 : 1000 dilution), Hand1 (ABCAM, 1 : 1000 dilution), GATA-3, GATA-4 and HMGA2 (all Cell Signaling Technology, all 1 : 1000 dilution). All primary antibodies were incubated overnight at 4 °C and detected using anti-mouse- or anti-rabbit-horseradish peroxidase (HRP)-conjugated antibodies (Cell Signaling Technology). Chemiluminescence solution (GE Healthcare) was used to detect secondary antibodies on film. Where necessary, blots were stripped and reprobed.

Hyde K., Sultana N., Tran A.C., Bileckaja N., Donald C.L., Kohl A., Stanton R.J, & Strang B.L. (2021). Limited replication of human cytomegalovirus in a trophoblast cell line. The Journal of General Virology, 102(11), 001683.

+ Open protocol

+ Expand

Comprehensive Antibody Inventory for Cardiac Research

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

The following primary antibodies were used for immunofluorescence and immunoblot analyses as well as coimmunoprecipitation experiments: Stat1 (Cell Signaling, 9172), p-Stat1 S727 (Cell Signaling, 9177), p-Stat1 Y701 (Cell Signaling, 9167), Smad2/3 (Cell Signaling, 5678); Smad3 and p-Smad3 (Cell Signaling [ref. 22 (link)]), SRF (Cell Signaling, 5147), Gata-4 (Cell Signaling, 36966 or Developmental Studies Hybridoma Bank [DSHB]; CRP-Gata-4-1A7, deposited to the DSHB by Protein Capture Reagents Program, produced by JHU/CDI), p-Histone H3 S10 (Cell Signaling, 3377), JAK1 (Cell Signaling, 3332), STAT3 (Cell Signaling, 9404), GAPDH (Santa Cruz Biotechnology, sc-32233), titin T11 (MilliporeSigma, T9030), sarcomeric myosin (DSHB; A4.1025), troponin-I (Cell Signaling, 4002), sarcomeric α-actinin-2 (clone EA-53, Mob 227-05, Diagnostic Biosystems), cardiac actin (Progen Biotechnik, Ac1–20.4.2), PTEN (Cell Signaling, 9188), FoxO1 (Cell Signaling, 2880), normal rabbit IgG (Santa Cruz Biotechnology, sc-2027), normal mouse IgG (Santa Cruz Biotechnology, sc-2025). Secondary fluorescence dye–linked or horseradish peroxidase–linked antibodies were obtained from Jackson Immunoresearch, DAKO, Cell Signaling or Santa Cruz Biotechnology. Fluorescently labeled phalloidin and DAPI was purchased from Molecular Probes (Life Technologies).

Lange S., Banerjee I., Carrion K., Serrano R., Habich L., Kameny R., Lengenfelder L., Dalton N., Meili R., Börgeson E., Peterson K., Ricci M., Lincoln J., Ghassemian M., Fineman J., del Álamo J.C, & Nigam V. (2019). miR-486 is modulated by stretch and increases ventricular growth. JCI Insight, 4(19), e125507.

+ Open protocol

+ Expand

Antibodies Used for Chromatin Analysis

Check if the same lab product or an alternative is used in the 5 most similar protocols

The antibodies used in this study were purchased from the following sources: Millipore (Billerica, MA): ASYM25 (09-814), HP1 (MAB3448), histone H3 (06-755), H3ac (06-599), H3K4me3 (07-473), H3K9me2 (07-441), H3K9me3 (07-442), H3K27me3 (07-449), H4 (05-858), H4ac (06-598), Prmt1 (07-404) and Sox2 (AB5603); Santa Cruz Biotech (Santa Cruz, CA): actin (sc-47778), c-Myc (sc-40, sc-789), GAPDH (sc-166574), Gata4 (sc-9053), Gata6 (sc-9055), GST (sc-138), mSin3a (sc-994), Oct4 (sc-5279) and p300 (sc-32244); Cell Signaling Technology (Danvers, MA): HDAC1 (2062), HDAC2 (2545s) and KLF4 (4038); Abcam (Cambridge, UK): H3R17me2 (ab412); Sigma (St. Louis, MO): FLAG (F3165); MBL (Japan): FLAG (PM020); Active Motif (Carlsbad, CA): H4R3me2a (39705); R&D Systems (Minneapolis, MN): KLF4 (AF3158); and Bethyl Laboratories (Montgomery, TX): Nanog (A300-397A).
A specific monoclonal antibody against Klf4-R396me2a was generated by Absea Biotechnology Ltd (Beijing, China) using synthesized peptide CGRRSWPRKRTATHT, corresponding to residues aa 387–402 of Klf4, as an antigen.

Zuo Z.Y., Yang G.H., Wang H.Y., Liu S.Y., Zhang Y.J., Cai Y., Chen F., Dai H., Xiao Y., Cheng M.B., Huang Y, & Zhang Y. (2022). Klf4 methylated by Prmt1 restrains the commitment of primitive endoderm. Nucleic Acids Research, 50(4), 2005-2018.

+ Open protocol

+ Expand

Comprehensive Protein Expression Analysis

Cited 1 time

Check if the same lab product or an alternative is used in the 5 most similar protocols

Western blot procedure was performed according to the method as previously described (14 (link)). The primary antibodies E-cadherin (BD Biosciences, catalog no: 610181 RRID:AB_397581), N-cadherin (BD Biosciences, catalog no: 610920 RRID:AB_398236), Vimentin (BD Biosciences, catalog no: 550513 RRID:AB _393716), iNOS (BD Biosciences, catalog 610332, RRID:AB_397722), β-actin (Sigma-Aldrich, catalog no: MAB1501 clone C4; catalog no: A2066 RRID:AB_476693), Cleaved caspase-9 (Cell Signaling Technology, catalog no: 9502 RRID:AB_2068621); (Cell Signaling Technology, catalog no: 7237 RRID:AB_10895832), Fibronectin [Cell Signaling Technology, catalog no: 26836 Fibronectin/FN1 (E5H6X) Rabbit mAb], Gata4 (Cell Signaling Technology, catalog no: 36966 RRID:AB_2799108), Lamin A/C (Cell Signaling Technology, catalog no: 4777, RRID:AB_10545756), Snail2 (Cell Signaling Technology, catalog no: 9585, RRID:AB_2239535), Twist1 (Cell Signaling Technology, catalog no: 69366, RRID:AB_2891135), Zeb2 [Santa Cruz Biotechnology, catalog no: sc-271984 (E-11)], CD45 (Miltenyi Biotec, catalog no: 130–115–938, RRID:AB_2751284), EpCam (Invitrogen, catalog no: MA5–13917 RRID:AB_11001308); (Novus Biologicals, catalog no: NBP2–27107), CRB2 (ThermoFisher Scientific, catalog no: PA5–25628, RRID:AB_2543128).

Du Q., Liu S., Dong K., Cui X., Luo J, & Geller D.A. (2022). Downregulation of iNOS/NO Promotes Epithelial–Mesenchymal Transition and Metastasis in Colorectal Cancer. Molecular Cancer Research, 21(2), 102-114.

+ Open protocol

+ Expand

Western Blot Analysis of Mef2C and Gata4

Check if the same lab product or an alternative is used in the 5 most similar protocols

Proteins were extracted in lysis buffer (62.5mM Tris pH7.4, 1% SDS, 1% protease inhibitor cocktail-I (Sigma)). Following extraction, proteins were subjected to SDS-PAGE and then transferred to nitrocellulose membranes (Bio-Rad). Membranes were blocked (5%w/v non-fat dry milk in TBS-Tween: 25mM Tris pH7.4; 137mM NaCl; 0.1%v/v Tween-20) and then incubated overnight at 4°C with Mef2C (Cell Signaling, Cat no 5030S) or Gata4 (Cell Signaling, Cat no 36966S) antibodies diluted (1:1000 dilution) in TBS-Tween containing 5%w/v BSA. Following washing in TBS-Tween, membranes were incubated in HRP-conjugate secondary antibodies (Cell Signaling) diluted (1:1000 dilution) in TBS-Tween containing 5%w/v non-fat dry milk for 1 hour at room temperature. Following washing with TBS-Tween, membranes were developed with the ECL-Plus system according to the manufacturer’s instructions (Amersham Biosciences). Bands were visualized with a G-Box (Syngene).

Dal-Pra S., Hodgkinson C.P, & Dzau V.J. (2019). Induced cardiomyocyte maturation: Cardiac transcription factors are necessary but not sufficient. PLoS ONE, 14(10), e0223842.

+ Open protocol

+ Expand

Characterization of C3A-iCSCs by Immunostaining

Check if the same lab product or an alternative is used in the 5 most similar protocols

C3A-iCSCs were passaged onto gelatin-coated dishes and incubated in DMEM containing 10% fetal bovine for another 7 days. The cells were then stained for GATA4 (Cell Signaling, Danvers), GFAP (Cell Signaling, Danvers), and BRY (Cell Signaling, Danvers) by immunofluorescence.

Han S., Guo J., Liu Y., Zhang Z., He Q., Li P., Zhang M., Sun H., Li R., Li Y., Zeng W., Liu J., Lian L., Gao Y, & Shen L. (2015). Knock out CD44 in reprogrammed liver cancer cell C3A increases CSCs stemness and promotes differentiation. Oncotarget, 6(42), 44452-44465.

+ 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?

Revolutionizing how scientists
search and build protocols!