Vdac 4866

Manufactured by Cell Signaling Technology

Sourced in United States

VDAC (4866) is a primary antibody that recognizes the Voltage-Dependent Anion Channel (VDAC) protein. VDAC is a highly conserved, abundant, and essential channel protein located in the outer mitochondrial membrane that regulates the exchange of metabolites, ions, and other molecules between the cytoplasm and the mitochondria.

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6 protocols using vdac 4866

Multimodal Mitochondrial Function Analysis

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Primary antibodies used: Akt1 (2938), Akt2 (5239), Akt1/2 (9272), GSK-3α/β (5676), LC3 (4108S), Lamin A/C (2032), NFR1 (12381), PGC-1α (2178), phospho-p38 MAPK (9215), phospho-Akt1 (9018), phospho-Akt2 (8599), phospho-DRP1 (Ser616) (4494), phospho-GSK-3α/β (9331), Parkin (2132S), Tfam (7495S), VDAC (4866) (Cell Signaling); β-actin (A5441) (Sigma); PINK1 (ab23707), Mfn-2 (ab124773) (Abcam); Drp1 (611113) (BD Biosciences); phospho-PGC-1α (S571) (AF6650) (R&D Systems); p38 (sc-7972) and Tom20 (sc-11415) (Santa Cruz); SPC (AB3786) (Millipore).

Larson‐Casey J.L., Gu L., Davis D., Cai G., Ding Q., He C, & Carter A.B. (2021). Post‐translational regulation of PGC‐1α modulates fibrotic repair. The FASEB Journal, 35(6), e21675.

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Mitochondrial Dynamics and Signaling Pathways

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Primary antibodies used: Akt1 (2938), Akt2 (5239), Akt1/2 (9272), GSK‐3α/β (5676), LC3 (4108S), Lamin A/C (2032), NFR1 (12381), PGC‐1α (2178), phospho‐p38 MAPK (9215), phospho‐Akt1 (9018), phospho‐Akt2 (8599), phospho‐DRP1 (Ser616) (4494), phospho‐GSK‐3α/β (9331), Parkin (2132S), Tfam (7495S), VDAC (4866) (Cell Signaling); β‐actin (A5441) (Sigma); PINK1 (ab23707), Mfn‐2 (ab124773) (Abcam); Drp1 (611113) (BD Biosciences); phospho‐PGC‐1α (S571) (AF6650) (R&D Systems); p38 (sc‐7972) and Tom20 (sc‐11415) (Santa Cruz); SPC (AB3786) (Millipore).

Larson‐Casey J.L., Gu L., Davis D., Cai G., Ding Q., He C, & Carter A.B. (2021). Post‐translational regulation of PGC‐1α modulates fibrotic repair. The FASEB Journal, 35(6), e21675.

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Multiprotein Complex Purification Protocol

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BCLb #ab45412 and GFP #ab6673 (Abcam); VDAC #4866 (Cell Signaling); IGF1R #3018 (Cell Signaling), ESYT2 #HPA002132 (SIGMA), S5a/PSMD4 #12441 (Cell Signaling), BAG6 #A302-039A-T (Bethyl), MYC #F2512 (Santa cruz), Tubulin #B512, FLAG M2 conjugated agarose beads, FLAG poly-clonal #F7425 (Sigma); UBQLN poly-clonal was made by inoculating rabbits with a peptide specific to UBQLN1 (Yenzym Antibodies LLC); HA affinity matrix and HA #3F10 (Roche).

Kurlawala Z., Shah P.P., Shah C, & Beverly L.J. (2017). The STI and UBA domains of UBQLN1 are critical determinants of substrate interaction and proteostasis. Journal of cellular biochemistry, 118(8), 2261-2270.

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Quantifying Protein Expression in Cellular Immunostaining

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Cells were sorted, seeded on coverslips, fixed, immunostained, and the intensity quantified as described in Iborra and Buckle (2008) (link).
In Figures 1, 2, and 3, the protein antibodies used were MTOR(2983), PFKP(12746), FASN(3180), HK2(2867), LDHA(2012), PRPS6(4858) (Cell Signaling); SF3B4(AB66659), HNRNPU(AB20666), NFE2L2(AB31163), EPRS(AB31531), RRS(AB31537), HK4me3(AB8580) (Abcam); HNRNPA0(SC-16509), EP300(SC-584), TBP(SC-56794), GTF2F(SC-235), PKM2(SC-365684), UBTF(SC-13125), RNA Pol II(RPB1)(SC-9001) (Santa Cruz Biotechnologies); CLU(HPA000572), SRSF2(HPA049905), MCL1(HPA008455), TUBB(T8328), G6PD(HPA000834), PDIA2(HPA051692), GAPDH(G9545), PTBP1(WH0005725M1) (Sigma-Aldrich); GLUT1(AB1340) (Chemicon); BrU(MD5010) (Caltag); H4K20me2(39173), H3K36me2(39255), H4K16(39929), H3K27me3(61017) (Active Motifs); and PABP (kindly provided by Dr. Amelia Nieto).
For Figure 5B, the 24 antibodies used for protein quantification were HK2(2867), HDAC1(5356), LDHA(2012), RPS6(2217), EIF4B(13088), VDAC(4866), PFKP(12746) (Cell Signaling); G6PD(HPA000834), MCL1(HPA008455), TOMM22(HPA003037), CLU(HPA000572), PTBP1(WH0005725M1) (Sigma-Aldrich); NPM1(SC-6013), NCL(SC-8031), UBTF(SC-13125), PKM2(SC-365684), GTF2F(SC-235), BCL2(SC-492) (Santa Cruz Biotechnology); NFE2L2(AB31163), SF3B4(AB66659), IMMT(AB110329) (Abcam); GLUT1(AB1340) (Chemicon); EIF5(611976), EIF6(611120) (BD).

Guantes R., Rastrojo A., Neves R., Lima A., Aguado B, & Iborra F.J. (2015). Global variability in gene expression and alternative splicing is modulated by mitochondrial content. Genome Research, 25(5), 633-644.

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Apoptosis Signaling Pathway Profiling

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Primary antibodies used: Akt1 (2938), Bad (9239), Lamin A/C (2032), Bax (2772), phospho-Akt1 (9018), phospho-Bad (Ser136) (4366), VDAC (4866) (Cell Signaling); β-actin (A5441) (Sigma); p38 (sc-7972) (Santa Cruz); NOX4 (NB110-58849) (Novus), NOX4 (109225) (Abcam); Bak (A0404) (Abclonal).

Larson-Casey J.L., Gu L., Kang J., Dhyani A, & Carter A.B. (2021). NOX4 regulates macrophage apoptosis resistance to induce fibrotic progression. The Journal of Biological Chemistry, 297(1), 100810.

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Immunoblot Analysis of Mitochondrial Proteins

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Immunoblot analysis was performed as we described previously30 (link). Rabbit polyclonal anti-ANT-1 antibody and rabbit monoclonal anti-β-actin antibody were from Sigma-Aldrich (St Louis, MO, USA, SAB2105530, 1:1,000) and Cell Signaling (Danvers, MA, USA, 4970, 1:5,000), respectively. Rabbit polyclonal anti-ANT2 antibody was generated by immunization with KLH-conjugated N-terminus ANT2 peptide (NH-TDAAVSFAKDFLAG-COOH) at Lampire Biological Laboratories (Ottsville, PA, USA) and purified with protein G (GE Healthcare, Buckinghamshire, UK) and antigen-conjugated columns. The ANT2 antibody was diluted to 1:200 as being used. COX IV (4850, 1:1,000) and VDAC (4866, 1:1,000) antibodies were from Cell Signaling (Danvers, MA, USA). Tom20 antibody was from Santa Cruz Biotechnology (sc-11415, 1:500), Catalase and SOD2 antibodies were from Sigma (St. Louis, MO, USA, C0979, 1:1,000) and Abcam (Cambridge, UK, ab16956, 1:1,000), respectively. Uncropped scans of the critical immunoblots are shown in Supplementary Fig. 6.

Cho J., Zhang Y., Park S.Y., Joseph A.M., Han C., Park H.J., Kalavalapalli S., Chun S.K., Morgan D., Kim J.S., Someya S., Mathews C.E., Lee Y.J., Wohlgemuth S.E., Sunny N.E., Lee H.Y., Choi C.S., Shiratsuchi T., Oh S.P, & Terada N. (2017). Mitochondrial ATP transporter depletion protects mice against liver steatosis and insulin resistance. Nature Communications, 8, 14477.

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