Anti-PGAM5 antibody [EPR27145-82]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PGAM5 antibody [EPR27145-82] (AB308449)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PGAM5 KO HeLa (PGAM5 knockout human cervix adenocarcinoma epithelial cell) (ab265141) cells labelling PGAM5 with ab308449 at 1/250 (2.12 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing mitochondrial staining in parental HeLa, and no staining in PGAM5 KO HeLa cell line. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). anti-COX IV mouse monoclonal antibody - Mitochondrial Marker was used to counterstain tubulin at 1/1000 1ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-PGAM5 antibody [EPR27145-82] (AB308449)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized PGAM5 KO HeLa (PGAM5 knockout human cervix adenocarcinoma epithelial cell) (ab265141) (Left) / Parental HeLa (Right) cells labelling PGAM5 with ab308449 at 1/500 dilution (0.1 ug)/Red (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/5000 dilution was used as the secondary antibody.

• IP

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Immunoprecipitation - Anti-PGAM5 antibody [EPR27145-82] (AB308449)

PGAM5 was immunoprecipitated from 0.35 mg HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate with ab308449 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab308449 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 2 : ab308449 IP in HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab308449 in HeLa whole cell lysate

Blocking and dilution buffer and concentration : 5% NFDM/TBST.

Exposure time : 180 seconds

All lanes:

Immunoprecipitation - Anti-PGAM5 antibody [EPR27145-82] (ab308449) at 1/1000 dilution

All lanes:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 10 µg

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

Observed band size: 32 kDa

false

Exposure time: 180s