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Ab5245

Manufactured by Abcam
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Ab5245 is a lab equipment product from Abcam. It is a device designed for use in laboratory settings, but a detailed description of its core function is not available at this time.

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Lab products found in correlation

A7811, by Merck Group (2 mentions) Alexa fluor 647 phalloidin, by Thermo Fisher Scientific (2 mentions) Bovine serum albumin (bsa), by Merck Group (2 mentions) Triton x 100, by Merck Group (2 mentions) Sp8 x inverted confocal microscope, by Leica (2 mentions) Alexa fluor 488 goat anti mouse, by Thermo Fisher Scientific (1 mentions) Pierce ecl system, by Thermo Fisher Scientific (1 mentions) Ab236101, by Abcam (1 mentions) Nb100 2219, by Novus Biologicals (1 mentions) Sc 9053, by Santa Cruz Biotechnology (1 mentions) Hrp conjugated secondary antibody, by Bio-Rad (1 mentions) Nb100 479, by Novus Biologicals (1 mentions)

3 protocols using ab5245

1

Cardiomyocyte Hydrogel Immunofluorescence Staining

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Cardiomyocyte encapsulating hydrogels were fixed with 4% paraformaldehyde for 20 min at RT and washed with PBS. Gels were blocked in 1% BSA (Sigma) and permeabilized with 0.3% Triton-X 100 (Sigma) for min 2 h at RT. Cells were stained with mouse anti-α-actinin (sarcomeric; A7811, Sigma) and rabbit anti-GATA-4 (ab5245, Abcam) for overnight at 4 °C. Samples were washed and stained for secondary goat anti-mouse Alexa Fluor 488 and goat anti-rabbit Alexa Fluor 568 antibodies (Life Technologies) for 2 h at RT. Actin cytoskeleton was visualized with Alexa Fluor 647 phalloidin (Life Technologies) and DAPI was used as nuclear counter staining. After washing, hydrogels were visualized with Leica SP8 X inverted confocal microscope (Leica).
Tasoglu S., Yu C.H., Gungordu H.I., Guven S., Vural T, & Demirci U. (2014). Guided and magnetic self-assembly of tunable magnetoceptive gels. Nature Communications, 5, 4702.
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2

Immunofluorescence Staining of Cardiomyocyte Hydrogels

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Cardiomyocyte encapsulating hydrogels were fixed with 4% paraformaldehyde for 20 minutes at room temperature (RT) and washed with PBS. Gels were blocked in 1% BSA (Sigma) and permeabilized with 0.3% Triton-X 100 (Sigma) for min 2h at RT. Cells were stained with mouse anti-α-actinin (sarcomeric) (A7811, Sigma) and rabbit anti-GATA-4 (ab5245, Abcam) for overnight at 4°C. Samples were washed and stained for secondary goat anti-mouse Alexa Fluor® 488 and goat anti-rabbit Alexa Fluor® 568 antibodies (Life Technologies) for 2h at RT. Actin cytoskeleton was visualized with Alexa Fluor® 647 phalloidin (Life Technologies) and DAPI was used as nuclear counter staining. After washing hydrogels were visualized with Leica SP8 X inverted confocal microscope.
Tasoglu S., Yu C.H., Gungordu H.I., Guven S., Vural T, & Demirci U. (2014). Guided and magnetic self-assembly of tunable magnetoceptive gels. Nature Communications, 5, 4702.
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3

Cardiac Protein Detection and Quantification

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For GATA4, pGATA4 and BNP thirty μg and for MEF2C hundred μg of total protein from heart tissue was lysed in EBC + urea buffer (50 mM Tris-HCl pH 7.5, 150 mM NaCl, 0.5% NP-40, 3 M urea) resolved by SDS-PAGE, transferred to PVDF membrane, blotted, and probed with the following primary antibodies: anti-GATA4 pSer105 (1:600 ab5245, Abcam), anti-GATA4 (1:1000 sc-9053, Santa Cruz Biotechnology), anti-BNP (1:500 ab236101, Abcam), anti-vinculin (1:200 V4505, Merck) and anti-MEF2C (1:7500 sc-313×, Santa Cruz Biotechnology) followed by a HRP-conjugated secondary antibody (1:5000; Bio-Rad). For blotting of HIF1α and HIF-P4H-2 hundred ug of total protein from heart tissue lysed using EBC + urea lysis buffer and was resolved by SDS-PAGE, transferred with nitrocellulose membrane, blotted and probed with anti-HIF1α (1:500, NB100-479 Novus Biologicals) and anti-HIF-P4H-2 (1:500, Novus Biologicals NB100-2219)primary antibodies followed by HRP-conjugated secondary antibody. The Pierce ECL system (Ther-moScientific) was used for detection. Western blot images were quantified with fiji (ImageJ) software. For original blots see (Supplementary Fig. S1-S8).
Röning T., Magga J., Laitakari A., Halmetoja R., Tapio J., Dimova E.Y., Szabo Z., Rahtu-Korpela L., Kemppi A., Walkinshaw G., Myllyharju J., Kerkelä R., Koivunen P, & Serpi R. (2022). Activation of the hypoxia response pathway protects against age-induced cardiac hypertrophy. Journal of molecular and cellular cardiology, 164.
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