Anti-HMGCS1 antibody - C-terminal

• IP

Supplier Data

Immunoprecipitation - Anti-HMGCS1 antibody - C-terminal (AB194971)

Detection of HMGCS1 in immunoprecipitates of 293T whole cell lysate (prepared using NETN lysis buffer; 1 mg for IP, 20% of IP loaded) using ab194971 at 6 μg/mg lysate for IP and at 0.1 μg/ml for subsequent Western blot detection.

Detection : Chemiluminescence with an exposure time of 30 seconds.

All lanes:

Immunoprecipitation - Anti-HMGCS1 antibody - C-terminal (ab194971)

Predicted band size: 57 kDa

false

• WB

Supplier Data

Western blot - Anti-HMGCS1 antibody - C-terminal (AB194971)

Lysates prepared using NETN lysis buffer.

All lanes:

Western blot - Anti-HMGCS1 antibody - C-terminal (ab194971) at 0.1 µg/mL

Lane 1:

HeLa whole cell lysate at 50 µg

Lane 2:

293T whole cell lysate at 50 µg

Lane 3:

Jurkat whole cell lysate at 50 µg

Lane 4:

TCMK-1 whole cell lysate at 50 µg

Lane 5:

NIH/3T3 whole cell lysate at 50 µg

Predicted band size: 57 kDa

true

Exposure time: 30s

• WB

CiteAb

Western blot - Anti-HMGCS1 antibody - C-terminal (AB194971)

HMGCS1 western blot using anti-HMGCS1 antibody - C-terminal ab194971. Publication image and figure legend from Kühl, I., Miranda, M., et al., 2017, Elife, PubMed 29132502.

ab194971 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab194971 please see the product overview.

OXPHOS dysfunction leads to decreased cellular Q levels, but the enzymes of the mevalonate pathway are normal.(A) Scheme of the mevalonate and Q biosynthesis pathways, and OXPHOS complexes (Nuclear-encoded OXPHOS proteins are shown in yellow and mtDNA-encoded OXPHOS proteins in blue). Colored boxes : protein levels; red : increased, blue : decreased, grey : not detected or not quantified. (B) Heatmaps illustrating the fold-change protein levels of the Q biosynthesis pathway in alphabetical order of L/L, cre and L/L mouse hearts; blank boxes : not detected or not quantified proteins; p<0.05 in≥1 knockout strain. (C) Immunoblot of enzymes of the mevalonate pathway on total protein extracts from different L/L, cre and L/L hearts. Loading : tubulin. (D) Transcript levels of genes encoding enzymes of the mevalonate and coenzyme Q synthesis pathway in L/L, cre and L/L hearts. Normalization : B2M (beta-2-microglobulin). (E) Protein levels of OXPHOS complexes I-V and the downregulated Q biosynthesis enzymes at different time points in Lrpprc knockout mouse hearts compared to controls. The graph represents a mean log2 fold-change of all the proteins in that category. (F) Time point analysis of protein levels of enzymes of the Q biosynthesis pathway in Lrpprc knockout mouse hearts compared to controls. Adjusted p across time <0.05. (G) Quinone quantification (Q9 and Q10) in different L/L, cre and L/L mouse hearts. Error bars : ± SEM; *p<0.05, **p<0.01, ***p<0.001; two-tailed unpaired Student's t-test.10.7554/eLife.30952.031Figure 8—source data 1.qRT-PCR of genes encoding ubiquinone and mevalonate pathway enzymes in the five knockout mouse strains.10.7554/eLife.30952.032Figure 8—source data 2.Determination of coenzyme Q9 and 10.qRT-PCR of genes encoding ubiquinone and mevalonate pathway enzymes in the five knockout mouse strains.Determination of coenzyme Q9 and 10.Transcript levels of genes encoding for enzymes of the mevalonate and the Q synthesis pathways.(A) Heatmaps showing the fold-change transcript levels in alphabetical order of L/L, cre and L/L mouse hearts of the mevalonate and the Q biosynthesis pathways by RNA-Seq.; blank boxes : not detected or not quantified transcript; adjusted p<0.05 in≥1 knockout mouse line. (B) qRT-PCR of transcript levels of genes encoding enzymes of coenzyme Q synthesis pathway in different L/L, cre and L/L hearts, normalized to B2M (beta-2-microglobulin). All graphs represent mean ± SEM; *p<0.05, **p<0.01, ***p<0.001.

false