- PA5-28196 - Invitrogen Antibodies POLRMT antibody

Submitted references A deafness-associated mitochondrial DNA mutation caused pleiotropic effects on DNA replication and tRNA metabolism.

Meng F, Jia Z, Zheng J, Ji Y, Wang J, Xiao Y, Fu Y, Wang M, Ling F, Guan MX
Nucleic acids research 2022 Sep 9;50(16):9453-9469

Mitochondrial maturation drives germline stem cell differentiation in Caenorhabditis elegans.

Charmpilas N, Tavernarakis N
Cell death and differentiation 2020 Feb;27(2):601-617

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Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Western Blot using POLRMT Polyclonal Antibody (Product # PA5-28196). Sample (30 µg of whole cell lysate). Lane A: HepG2. 5% SDS PAGE. POLRMT Polyclonal Antibody (Product # PA5-28196) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Western Blot using POLRMT Polyclonal Antibody (Product # PA5-28196). Sample (30 µg of whole cell lysate). Lane A: HepG2. 5% SDS PAGE. POLRMT Polyclonal Antibody (Product # PA5-28196) diluted at 1:1,000. The HRP-conjugated anti-rabbit IgG antibody was used to detect the primary antibody.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Immunofluorescent analysis of mtRNA polymerase in methanol-fixed Hep G2 cells using a mtRNA polymerase polyclonal antibody (Product # PA5-28196) at a 1:500 dilution.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
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Experimental details: Immunohistochemical analysis of paraffin-embedded human hepatoma, using mtRNA polymerase (Product # PA5-28196) antibody at 1:500 dilution. Antigen Retrieval: EDTA based buffer, pH 8.0, 15 min.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Figure 2. mtDNA replication analysis. ( A ) Schematic representation of fragments digested by Acc I, Dra I, Bcl I and Apa I, and hybridized with probes a , b , c and d, respectively. ( B ) Replication intermediates in specific DNA regions. Eight micrograms of mtDNAs from various cell lines were digested with Acc I, Dra I, Bcl I and Apa I, and the resultant fragments were separated in a 0.4% agarose gel. A gel slice was then cut, embedded into a 1% agarose gel and subjected to a second-dimension gel electrophoresis. The 2D gel is then blotted and hybridized with DIG labeled probes a , b , c and d, respectively. ( C ) Schematic representation of the cleaved replication bubble and Y arc formed by mtDNA replication intermediates. ( D ) South-western analysis of mtDNA replication. BrdU incorporation into mtDNA was analyzed 24 h post-injection and bands were quantified from the same blot. The nuclear 18S rRNA gene was probed to control for loading. Representative bands are shown for clarity of comparison. ( E ) Western blot analysis of proteins related to mtDNA replication: PolgammaA, Twinkle, SSBP1, POLRMT and TFAM with beta -actin as a loading control in the mutant cell lines and control cell lines. ( F ) Quantification of proteins. Average relative values of PolgammaA, Twinkle, SSBP1, POLRMT and TFAM were normalized to the average values of beta -actin in various cell lines. The values for the latter are expressed as percentages of the average values for the control cell lines.

Supportive validation

Submitted by: Invitrogen Antibodies (provider)
Main image
Experimental details: Fig. 8 Mammalian stem cell differentiation upon LIF removal is accompanied by an increase in POLRMT expression. Elevated POLRMT expression is observed in cells with increased cytoplasmic OCT-4 abundance (arrowheads). In contrast, adjacent areas with increased nuclear OCT-4 abundance (stars) display lower POLRMT expression. Images were acquired using a x40 objective lens. Scale bars, 20 mum

PA5-28196

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