PA5-32132

antibody from Invitrogen Antibodies

Targeting: OXSM CEM1, FASN2D, FLJ20604, KS

Western blot (Data presented on provider website)

Immunohistochemistry (Data presented on provider website)

Other assay (Supportive data in Antibodypedia)

Antibody Data

Product number

PA5-32132

Provider

Invitrogen Antibodies

Product name

OXSM Polyclonal Antibody

Antibody type

Polyclonal

Antigen

Recombinant protein fragment

Description

Recommended positive controls: HeLa. Predicted reactivity: Mouse (88%), Rat (88%), Bovine (84%). Store product as a concentrated solution. Centrifuge briefly prior to opening the vial.

Reactivity

Human, Mouse, Rat

Host

Rabbit

Isotype

IgG

Vial size

100 µL

Concentration

1 mg/mL

Storage

Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.

References

Submitted references

Mitochondrial fatty acid synthesis coordinates oxidative metabolism in mammalian mitochondria.

Nowinski SM, Solmonson A, Rusin SF, Maschek JA, Bensard CL, Fogarty S, Jeong MY, Lettlova S, Berg JA, Morgan JT, Ouyang Y, Naylor BC, Paulo JA, Funai K, Cox JE, Gygi SP, Winge DR, DeBerardinis RJ, Rutter J
eLife 2020 Aug 17;9

Other assay

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 1. MtFAS is an essential pathway in mammalian skeletal myoblasts but does not contribute to synthesis of major cellular lipids. ( A ) Schematic of the mitochondrial fatty acid synthesis pathway and downstream lipoic acid synthesis. ( B ) Crude isolated mitochondrial fractions from duplicate single cell clones of Mcat , Oxsm , and Mecr mutants, compared with GFP control clonal cell lines, were separated via SDS-PAGE and immunoblotted for the indicated targets. *=Lipoic acid band (reprobe of earlier blot). #=non specific bands C . GFP control and Oxsm mutant cells were infected with retroviral control plasmid (pCtrl) or a plasmid expressing Oxsm off the CMV promoter (pOxsm), plated at equal densities in normal growth medium with either 4.5 g/L glucose or 10 mM galactose and grown for 3 or 4 days, respectively, then stained with crystal violet. ( D ) Whole cell lysates from stable cell lines generated by infecting Oxsm mutant cells (OxsmDelta) or GFP controls with shRNA constructs targeting FASN (shFASN) or scramble control (shScramble) were separated by SDS-PAGE and immunoblotted for FASN, lipoylated proteins, or tubulin. ( E-F ) Stable cell lines created in ( D ) were incubated with U 13 C-glucose for the indicated number of doublings, harvested, lipids extracted, and analyzed via LC-MS. Shown are quantitation of m+2 isotopologues for two representative phospholipid species, PC 16:0_16:0 and PC 16:0_18:0. +=p < 0.001, ++=p < 0.0001, error bars are SEM. Figure 1--figure

Supportive validation

Submitted by

Invitrogen Antibodies (provider)

Main image

Experimental details

Figure 5. MtFAS mutant phenotypes are not recapitulated by loss of lipoic acid alone. ( A ) Immunoblot for the indicated proteins in whole cell lysates from Lipt1 mutant and control cell lines. ( B ) In triplicate experiments, cells of each of the indicated clones were seeded in eight wells of a 96-well seahorse plate and allowed to adhere overnight, then equilibrated and treated with the indicated drugs following standard mitochondrial stress test protocols from the manufacturer to determine Oxygen Consumption Rate (OCR). #=p < 0.01, +=p < 0.001, ++=p < 0.0001 all comparisons are to GFP control, error bars are SEM. ( C ) Mitochondrial lysates generated from the indicated cell lines by differential centrifugation were normalized for total protein by BCA assay, incubated with 1% digitonin, then separated by BN-PAGE and immunoblotted with the indicated antibodies. ( D ) Four biological replicates from mtFAS mutant cell lines or GFP control were grown under standard proliferative conditions and harvested for steady-state metabolomics analysis by LC-MS. Shown are relative pool sizes for the indicated metabolites. *=p < 0.05, error bars are SD. ( E ) Four biological samples of the indicated genotype were labeled for 24 hr with U 13 Cglutamine, harvested, and analyzed via GC-MS for the indicated metabolites and their isotopologues. *=p < 0.05, #=p < 0.01, +=p < 0.001, error bars are SD. Figure 5--figure supplement 1. Expression of mtFAS proteins is unchanged in Lipt1 mutant cell li