Immunoblotting Protein Detection Protocol

Immunoblotting was performed as previously described^{54 (link)}. Actin (C4) and PGC1α (H-300) antibodies for immunoblots were obtained from Santa Cruz Biotechnology, p-AKT (S473) (D9E), pS6 (S235/236) (D57.2.2E), BNIP3 #3769, LCK #2752, p-Tyr-100 #9411 from Cell Signaling, and CV- ATP5A, CIII- UQCRC2, CII- SDHB from Total Rodent OXPHOS Rodent WB antibody cocktail from Abcam. Immunoblots were detected via standard secondary detection and chemiluminescent exposure to film. Digitally captured films were analyzed densitometrically by ImageJ software.

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Scharping N.E., Rivadeneira D.B., Menk A.V., Vignali P.D., Ford B.R., Rittenhouse N.L., Peralta R., Wang Y., Wang Y., DePeaux K., Poholek A.C, & Delgoffe G.M. (2021). Mitochondrial stress induced by continuous stimulation under hypoxia rapidly drives T cell exhaustion. Nature immunology, 22(2), 205-215.

Publication 2021

PGC1α (H-300) BNIP3 #3769 LCK #2752 p-Tyr-100 #9411 OXPHOS Rodent WB antibody cocktail

Actin Antibodies Antibody cocktail Immunoblots Pgc1α Rodent Sdhb

Corresponding Organization :

Other organizations : UPMC Hillman Cancer Center, Children's Hospital of Pittsburgh, University of Pittsburgh, Tsinghua University

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Variable analysis

independent variables

None explicitly mentioned

dependent variables

Protein expression levels of Actin, PGC1α, p-AKT (S473), pS6 (S235/236), BNIP3, LCK, p-Tyr-100, CV-ATP5A, CIII-UQCRC2, CII-SDHB

control variables

None explicitly mentioned

controls

Positive controls: None specified
Negative controls: None specified

Annotations

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