Invitrogen

PMA1 Monoclonal Antibody (40B7)

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PMA1 Antibody in Western Blot (WB) — Figure 2. APOL1 expression levels and cell proliferation on non-fermentable media. (A) APOL1 inhibited yeast proliferation in non-fermentable media: HA tagged wild type and BH3-deleted (BH3) APOL1 expression was induced by galactose and cells were then shifted in liquid glycerol containing MGly media at T=0. Error bars represent standard errors on three independent experiments. (B) APOL1 protein expression levels after 15.5 h: Cell lysates were analyzed using immunoblotting with an anti-HA antibody. Loading control was made using immunoblotting analysis with an anti-Pma1 antibody. (C) APOL1 interfered with the respiratory competence. The capability of carbon sources to support proliferation was analyzed by plating cells (at 103 density) on solid media containing galactose or glycerol. Viable colonies on MGly plates were counted and compared with viable colonies under fermentable conditions. (D) APOL1 and BH3 deleted-APOL1 (BH3) proteins were expressed during the index of respiratory competence assay (days 1, 3, 5 and 7). Cell extracts taken at the indicated time points were analyzed using immunoblotting with the indicated antibodies. Loading control was made using immunoblotting analysis with an anti-Pma1 antibody. APOL1, apolipoprotein L 1; MGly, glycerol-containing medium; OD, optical density; T, time.

PMA1 Antibody in Western Blot (WB) — Figure 1. APOL1 expression levels, cell proliferation and intracellular ROS. (A) HA tagged wild type APOL1 and BH3-deleted APOL1 (BH3) expression was induced in cells from a galactose inducible vector and revealed by immunoblotting assay using an anti-HA antibody; cells transfected with the empty vector (pGAL1) were used as negative control; loading control was made by immunoblotting analysis using an anti-Pma1 antibody. (B) APOL1 had no effect on yeast proliferation as measured using clonogenic assay: Cells analyzed in panel A were grown on solid media containing glucose or galactose and counted. (C) APOL1 had no effect on yeast proliferation as measured using liquid growth assay: Cells analyzed in panel A were inoculated in liquid media containing either glucose or galactose and proliferation was assessed by OD measurement. (D) Cells were treated with the ROS detecting reagent (dihydroethidium) and analyzed by flow cytometry. (E) Quantification of the flow cytometry analysis performed in D. Data are presented as the average of three independent cultures. Error bars represent the standard error of the mean. APOL1, apolipoprotein L 1; ROS, reactive oxygen species; OD, optical density.

PMA1 Antibody in Western Blot (WB) — Figure 3. APOL1 expression levels and mitochondrial membrane depolarization. (A) APOL1 depolarized the mitochondrial membrane. Cells were incubated with TMRE; total fluorescent and non-fluorescent cells were analyzed using flow cytometry assays and the percentage of non-fluorescent cells in the population was plotted. Experiments were performed in liquid MGal media. Error bars represent standard errors of the mean in three independent experiments. (B) Simultaneously with flow cytometry assays, cell lysates were prepared and analyzed using immunoblotting with an anti-HA antibody. Loading control was made using immunoblotting analysis with an anti-Pma1 antibody. (C) Cells were incubated with TMRE and analyzed using flow cytometry. FCCP was used as a positive control. P<0.001 (***) was considered statistically significant. APOL1, apolipoprotein L 1; TMRE, tetramethylrhodamine, ethyl ester; MGal, galactose-containing medium; FCCP, carbonilcyanide p-triflouromethoxyphenylhydrazone.

PMA1 Antibody in Western Blot (WB) — Figure 4. APOL1 subcellular localization. Extracts taken from cells transformed with the pGAL-1 vector expressing wild type APOL1 or the empty vector were fractionated. The UM and EM fractions are shown. These fractions were analyzed using western blotting with the following antibodies: Anti-HA, anti-Dpm1, anti-porin, anti-Pma1 and anti-PGK1. The UM fraction contains the endoplasmic reticulum, plasma membrane and mitochondrial fragments. The EM fraction is enriched for intact mitochondria. The figure shows non-adjacent bands from the same blot that were cut, adjusted and joined in order to avoid including non-relevant wells in the results. APOL1, apolipoprotein L 1; UM, unpurified mitochondria; EM, enriched mitochondria.

Product Details

MA1-91567

Applications	Tested Dilution	Publications
Western Blot (WB)	1:10,000	View 2 publications 2 publications
Immunocytochemistry (ICC/IF)	1:1,000-1:5,000	-

Product Specifications
Species Reactivity	Yeast
Published species	Yeast
Host/Isotype	Mouse / IgG
Class	Monoclonal
Type	Antibody
Clone	40B7
Immunogen	Yeast membrane preps.
Conjugate	Unconjugated
Form	Liquid
Concentration	Conc. Not Determined
Storage buffer	tissue culture supernatant
Contains	10mM sodium azide
Storage conditions	Store at 4°C short term. For long term storage, store at -20°C, avoiding freeze/thaw cycles.
RRID	AB_1959827

Product Specific Information

The predicted molecular weight by western blot is 100 kDa.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

Product References

Human apolipoprotein L1 interferes with mitochondrial function in Saccharomyces cerevisiae.

Molecular medicine reports

Chidiac M,Daher J,Boeckstaens M,Poelvoorde P,Badran B,Marini AM,Khalaf R,Vanhamme L

MA1-91567 was used in Western Blotting to present evidence which suggests that APOL1 could be a physiological regulator of mitochondrial function.

Tue Sep 01 00:00:00 EDT 2020

Human apolipoprotein L1 interferes with mitochondrial function in Saccharomyces cerevisiae.

Molecular medicine reports

Chidiac M,Daher J,Boeckstaens M,Poelvoorde P,Badran B,Marini AM,Khalaf R,Vanhamme L

MA1-91567 was used in Western Blotting to present evidence which suggests that APOL1 could be a physiological regulator of mitochondrial function.

Tue Sep 01 00:00:00 EDT 2020

Identification of residues involved in homodimer formation located within a β-strand region of the N-terminus of a Yeast G protein-coupled receptor.

Journal of receptor and signal transduction research

Uddin MS,Kim H,Deyo A,Naider F,Becker JM

MA1-91567 was used in western blot to investigate the structure-function relationship of a yeast G protein-coupled receptor

Sun Apr 01 00:00:00 EDT 2012

Bioinformatics

Protein Aliases: H(+)-exporting P2-type ATPase PMA1; Plasma membrane ATPase 1; Proton pump 1

Gene Aliases: KTI10; PMA1; YGL008C

UniProt ID: (Baker's yeast) P05030

Entrez Gene ID: (Baker's yeast) 852876

Molecular Function: primary active transporter

ATP biosynthetic process transport ion transport regulation of pH proton transport regulation of intracellular pH transmembrane transport hydrogen ion transmembrane transport proteasome storage granule assembly

Process(es)

nucleotide binding ATP binding hydrogen-exporting ATPase activity, phosphorylative mechanism hydrolase activity ATPase activity metal ion binding