Immunofluorescence staining from cells in 12-well plates was performed as previously described35 (link). Primary antibodies used included anti-MCT4 (19-mer peptide sequence CKAEPEKNGEVVHTPETSV-cooh affinity purified rabbit antibody; YenZym Antibodies), anti-GLB1 (ab96239; Abcam), anti-HMGB1 (NB100–2322; Novus Biologicals), and anti-MCT1 (19-mer peptide sequence CSPDQKDTEGGPKEEESPV-cooh affinity purified rabbit antibody; YenZym). Anti-rabbit AlexaFluor 568 (A11036; Invitrogen) secondary antibody was used. Nuclear counterstaining was performed with DAPI. Images were collected with a 40x objective and 1.5 or 2x zoom, when indicated, using a Nikon A1R confocal microscope.
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Immunofluorescence Staining of Cellular Proteins
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Antibodies
Antibody
Biologicals
Cells
Confocal microscope
Dapi
Glb1
Hmgb1
Immunofluorescence
Mct1
Novus
Peptide
Rabbit
Corresponding Organization : Thomas Jefferson University
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Variable analysis
independent variables
- Anti-MCT4 (19-mer peptide sequence CKAEPEKNGEVVHTPETSV-cooh affinity purified rabbit antibody; YenZym Antibodies)
- Anti-GLB1 (ab96239; Abcam)
- Anti-HMGB1 (NB100–2322; Novus Biologicals)
- Anti-MCT1 (19-mer peptide sequence CSPDQKDTEGGPKEEESPV-cooh affinity purified rabbit antibody; YenZym)
- Immunofluorescence staining from cells in 12-well plates
- Anti-rabbit AlexaFluor 568 (A11036; Invitrogen) secondary antibody
- Nuclear counterstaining with DAPI
- Positive control: Not specified
- Negative control: Not specified
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