- PA5-17025 - Invitrogen Antibodies MYH9 antibody

Submitted references A resilient formin-derived cortical actin meshwork in the rear drives actomyosin-based motility in 2D confinement.

Ramalingam N, Franke C, Jaschinski E, Winterhoff M, Lu Y, Brühmann S, Junemann A, Meier H, Noegel AA, Weber I, Zhao H, Merkel R, Schleicher M, Faix J
Nature communications 2015 Sep 29;6:8496

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Myosin IIa in extracts from Hela and HT-29 cells using Myosin IIa polyclonal antibody (Product # PA5-17025).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western blot analysis of Myosin IIa in extracts from COS cells, mock transfected or transfected with nonmuscle EGFP-myosin IIa or IIb using Myosin IIa polyclonal antibody (Product # PA5-17025).

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunofluorescent analysis of Myosin IIa in HeLa cells using a Myosin IIa polyclonal antibody (Product # PA5-17025) (green) showing colocalization with actin filaments that have been labeled with a fluorescent red phalloidin. DNA is labeled using a fluorescent blue dye.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 6 The ForA motility phenotypes are myosin II dependent. ( a ) Elimination of ForA in mhcA - cells was confirmed by immunoblot using myosin-II-specific antibodies. Porin served as a loading control. ( b ) Quantitative comparison of random cell migration of mhcA - and mhcA - / forA - cells in unconfined environments revealed no statistical difference. n , number of tracked cells. NS, not significant (Mann-Whitney U -test). ( c ) Radar plots of 20 cell trajectories each taken from representative samples. Scale bar, 40 mum. ( d ) Still images from time-lapse phase-contrast movies refer to Supplementary Movie 8 and illustrate that both cell lines are unable to migrate under agar. However, only mhcA - / forA - cells were able to form protrusion along their periphery (white arrowheads) in 2D confinement. Scale bar, 20 mum. ( e ) Kymograph analyses of mhcA - and mhcA - / forA - cells, respectively, that were selected from the movies depicted in d . Kymographs were generated along the indicated lines. Horizontal scale bar, 100 s. Vertical scale bar, 10 mum.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Figure 9 Subcellular localization of IQGAP1 and active mDia1 in B16-F1 cells. ( a ) B16-F1 melanoma cells, ectopically expressing EGFP-tagged IQGAP1, were plated on laminin-coated glass coverslips, fixed and labelled with Alexa488-conjugated nanobodies (green) and rhodamine phalloidin to visualize filamentous actin (red). Note the enrichment of IQGAP1 in the rear. ( b ) Subcellular localization of ectopically expressed EGFP-tagged active mDia1 in B16-F1 cells migrating on laminin. The cells were processed as described in a , but were additionally stained for myosin IIA. Active mDia1 was excluded from the leading edge and other actin-rich structures, with the exception of its prominent cortex localization in the trailing edge, where it colocalized together with cortical F-actin and myosin IIA. ( c ) mDia1 dynamics in repolarizing cells. Relocalization of active mDia1 in repolarizing B16-F1 cells to the new prospective end is indicated by white arrowheads and comparable to active ForA in Dictyostelium cells. Still images from time-lapse movies are shown. Time corresponds to min:s. Scale bars, 20 mum.

PA5-17025

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