14-9980-80

antibody from Invitrogen Antibodies

Targeting: GATA4

Western blot (Supportive data in Antibodypedia)

Immunohistochemistry (Recommended by provider)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

14-9980-80

Provider

Invitrogen Antibodies

Product name

Gata-4 Monoclonal Antibody (eBioEvan), eBioscience™

Antibody type

Monoclonal

Antigen

Other

Description

Description: The eBioEvan monoclonal antibody reacts with mouse and human Gata-4. In a western blot of C57Bl/6 and BALB/c mouse heart lysates, a single band is present, suggesting that this antibody does not cross-react with Gata-5 or Gata-6, which are also expressed in the heart, but with different molecular weights. The eBioEvan monoclonal antibody has been successfully used in flow cytometry on HEK 293T cells transiently transfected with either mouse Gata-4 or human Gata-4. Gata-4 is a member of the GATA family of transcription factors, which are characterized by a conserved DNA binding domain containing two zinc fingers. The mouse Gata-4 gene encodes a protein of 48 kDa. Studies show that Gata-4 plays an important role in controlling cardiomyocyte differentiation, proliferation and survival. Gata-4 is expressed during fetal development, as well as in the adult mouse heart. It also appears to modulate gene expression in the gut, liver, and gonads. Applications Reported: This eBioEvan antibody has been reported for use in immunoblotting (WB) and immunohistochemistry. Applications Tested: This eBioEvan antibody has been tested by western blot analysis of mouse heart lysates. As a starting dilution, it is recommended to use this purified antibody at 2 µg/mL for western blot analysis. The eBioEvan antibody has been tested on formalin-fixed paraffin embedded human tissue using low pH antigen retrieval and can be used at less than or equal to 5 µg/mL. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. Purity: Greater than 90%, as determined by SDS-PAGE. Aggregation: Less than 10%, as determined by HPLC. Filtration: 0.2 µm post-manufacturing filtered.

Reactivity

Human, Mouse

Host

Rat

Isotype

IgG

Antibody clone number

eBioEvan

Vial size

25 µg

Concentration

0.5 mg/mL

Storage

4° C

References

Submitted references

Imbalanced Activation of Wnt-/β-Catenin-Signaling in Liver Endothelium Alters Normal Sinusoidal Differentiation.

Koch PS, Sandorski K, Heil J, Schmid CD, Kürschner SW, Hoffmann J, Winkler M, Staniczek T, de la Torre C, Sticht C, Schledzewski K, Taketo MM, Trogisch FA, Heineke J, Géraud C, Goerdt S, Olsavszky V
Frontiers in physiology 2021;12:722394

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Naive stem cell blastocyst model captures human embryo lineage segregation.

Yanagida A, Spindlow D, Nichols J, Dattani A, Smith A, Guo G
Cell stem cell 2021 Jun 3;28(6):1016-1022.e4

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Diastolic dysfunction in a pre-clinical model of diabetes is associated with changes in the cardiac non-myocyte cellular composition.

Cohen CD, De Blasio MJ, Lee MKS, Farrugia GE, Prakoso D, Krstevski C, Deo M, Donner DG, Kiriazis H, Flynn MC, Gaynor TL, Murphy AJ, Drummond GR, Pinto AR, Ritchie RH
Cardiovascular diabetology 2021 Jun 1;20(1):116

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Spatiotemporal patterning of EpCAM is important for murine embryonic endo- and mesodermal differentiation.

Sarrach S, Huang Y, Niedermeyer S, Hachmeister M, Fischer L, Gille S, Pan M, Mack B, Kranz G, Libl D, Merl-Pham J, Hauck SM, Paoluzzi Tomada E, Kieslinger M, Jeremias I, Scialdone A, Gires O
Scientific reports 2018 Jan 29;8(1):1801

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Genetic Intersection of Tsix and Hedgehog Signaling during the Initiation of X-Chromosome Inactivation.

Del Rosario BC, Del Rosario AM, Anselmo A, Wang PI, Sadreyev RI, Lee JT
Developmental cell 2017 Nov 6;43(3):359-371.e6

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Dynamics of embryonic stem cell differentiation inferred from single-cell transcriptomics show a series of transitions through discrete cell states.

Jang S, Choubey S, Furchtgott L, Zou LN, Doyle A, Menon V, Loew EB, Krostag AR, Martinez RA, Madisen L, Levi BP, Ramanathan S
eLife 2017 Mar 15;6

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CDK14 expression is down-regulated by cigarette smoke in vivo and in vitro.

Pollack D, Xiao Y, Shrivasatava V, Levy A, Andrusier M, D'Armiento J, Holz MK, Vigodner M
Toxicology letters 2015 Apr 16;234(2):120-30

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Overexpression of aromatase associated with loss of heterozygosity of the STK11 gene accounts for prepubertal gynecomastia in boys with Peutz-Jeghers syndrome.

Ham S, Meachem SJ, Choong CS, Charles AK, Baynam GS, Jones TW, Samarajeewa NU, Simpson ER, Brown KA
The Journal of clinical endocrinology and metabolism 2013 Dec;98(12):E1979-87

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Lentiviral transduction of rat Sertoli cells as a means to modify gene expression.

Nicholls PK, Stanton PG, Rainczuk KE, Qian H, Gregorevic P, Harrison CA
Spermatogenesis 2012 Oct 1;2(4):279-284

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GATA-4 is a novel transcription factor expressed in endocardium of the developing heart.

Kelley C, Blumberg H, Zon LI, Evans T
Development (Cambridge, England) 1993 Jul;118(3):817-27

Western blot

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Anti-Human/Mouse Gata-4 Purified (2 µg/mL) was used in a western blot of SDS lysates. Lane 1: NIH-3T3, Lane 2: C57Bl/6 mouse heart, Lane 3: BALB/c mouse heart.

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Western blot analysis was performed on tissue extracts (30 µg lysate) of Mouse Heart (Lane 1), Mouse Ovary (Lane 2) and Mouse Brain (Lane 3). The blot was probed with Anti-GATA-4 Rat Monoclonal Antibody (Product # 14-9980-82, 1:1000 dilution) and detected by chemiluminescence using F(ab')2-Rabbit anti-Rat IgG (H+L) Secondary Antibody, HRP (Product # PA1-29927, 0.25 ug/ml, 1:4000 dilution). A 50 kDa band corresponding to GATA-4 was detected across the tissues tested. Additional band around 25kDa from circulating IgGs was observed in Mouse Heart and Mouse Ovary lysates but not in Mouse Brain lysate which is selectively permeable.

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

Fig. 1 Differences in the abundance of major non-myocyte cell classes in the diabetic heart. A Flow cytometry contour plot displaying gating of major non-myocyte cell types for quantification of cell type proportion (summarised in B). For full gating strategy see Additional file 1 : Figure S2. Endothelial cells (ECs; CD31 + ), resident mesenchymal cells (RMCs; CD31 - CD45 - ) and leukocytes (Leuks; CD45 + ). B Proportions of major cell types in non-diabetic (ND; n = 7) and diabetic (DM; n = 19) mouse hearts. Individual sample values are shown with mean +- SEM. C Immunohistochemical analysis of the abundance of RMCs in ND and diabetic mouse heart left ventricles. Left and middle panels show representative confocal micrographs of mouse heart tissue stained for PCM1 and GATA4. PCM1 + GATA4 + and PCM1 - GATA4 + nuclei correspond to nuclei of cardiomyocytes (CM) and RMCs respectively. Nuclei are counterstained with DAPI. Right panel (box-plot) summarises proportion of nuclei corresponding to RMCs in ND (n = 9) vs. DM (n = 10) enumerated from micrographs. Whiskers of box-and-whisker plot indicate max and min. (D) As for C, heart left ventricle sections were stained with DACH1 to identify nuclei corresponding to endothelial cells in ND (n = 10) and DM (n = 9) left ventricles. * P < 0.05, **** P < 0.0001 (Student's unpaired t -test). Scale bar = 100 uM

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 5 Hepatic endothelial Ctnnb1 overactivation causes loss of LSEC-associated genes and induction of CEC and brain EC genes. (A) IF staining of DAPI, LYVE1, and Emcn in the liver of 2- to 3-month-old female Ctnnb1 WT and Ctnnb1 OE - EC mice ( n = 4). Scale bar 100 mum. (B) Left panel: IF quantification of LYVE1 + area. Results are represented as mean +- SEM. ** p < 0.01. Right panel: qRT-PCR for LYVE1 of cDNA from Ctnnb1 OE - EC -LSECs compared to Ctnnb1 WT control LSECs ( n = 4). beta-Actin was used as housekeeping gene. * p < 0.05. (C) Bmp2, Hgf , Wnt2, Wnt9b mRNA RNAScope in situ hybridization assay of 2- to 3-month-old female Ctnnb1 WT and Ctnnb1 OE - EC mice liver sections ( n >= 3). Scale bar 100 mum. (D) Quantification of Bmp2, Hgf , Wnt2, Wnt9b mRNA RNAScope in situ hybridization assay. Results are represented as mean +- SEM. ns, not significant. * p < 0.05. (E) Left panel: IF staining of DAPI and GATA4 in the liver of 2- to 3-month-old female Ctnnb1 WT and Ctnnb1 OE - EC mice ( n = 4). Scale bar 100 mum. Right panel: qRT-PCR for Gata4 with cDNA from Ctnnb1 OE - EC -LSECs compared to Ctnnb1 WT control LSECs ( n = 4). beta-Actin was used as housekeeping gene. n.s., not significant. (F) qRT-PCR for Myc and Pdgfb with cDNA from Ctnnb1 OE - EC -LSECs compared with Ctnnb1 WT control LSECs ( n = 3). beta-Actin was used as housekeeping gene. * , p < 0.05; n.s., not significant. (G) qRT-PCR for CD34 with cDNA from Ctnnb1 OE - EC -LSECs compared with Ctnnb1 WT control LSEC