Anti-PGAM5 antibody

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-PGAM5 antibody (AB126534)

Immunocytochemical analysis of MCF7 (Human breast adenocarcinoma cell line) whole cells labeling PGAM5 in the mitochondria with ab126534 2 μg/ml.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGAM5 antibody (AB126534)

Immunohistochemical analysis of human colon tissue labeling PGAM5 in the granular cytoplasm of glandular cells with ab126534 at a 1/200 dilution.

Performed heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGAM5 antibody (AB126534)

Immunohistochemical analysis of human liver tissue labeling PGAM5 in the granular cytoplasm of hepatocytes with ab126534 at a 1/200 dilution.

Performed heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGAM5 antibody (AB126534)

Immunohistochemical analysis of human testis tissue labeling PGAM5 in the granular cytoplasm of cells in seminiferous ducts with ab126534 at a 1/200 dilution.

Performed heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGAM5 antibody (AB126534)

Immunohistochemical analysis of human tonsil tissue labeling PGAM5 in the granular cytoplasm of germinal center cells with ab126534 at a 1/200 dilution.

Performed heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• WB

Supplier Data

Western blot - Anti-PGAM5 antibody (AB126534)

Loading control : Anti-PPIB

Genetic validation in WB by siRNA knockdown.

All lanes:

Western blot - Anti-PGAM5 antibody (ab126534) at 0.4 µg/mL

Lane 1:

Control siRNA transfected into RT4 (Human urinary bladder cancer cell line) whole cell lysate

Lane 2:

siRNA probe #1 transfected into RT4 whole cell lysate

Lane 3:

siRNA probe #2 transfected into RT4 whole cell lysate

Predicted band size: 32 kDa

false

• WB

Supplier Data

Western blot - Anti-PGAM5 antibody (AB126534)

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

In Western blot, ab322207 was shown to bind specifically to PGAM5. Target of interest was observed at 32 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in PGAM5 knockout cell line (lane 2) (lane 2, knockout cell line ab265141 / knockout cell lysate ab257581).

Exposure time : Left panel : 180 seconds; right panel : 70 seconds

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PGAM5 antibody [RM1254] (<a href='/en-us/products/primary-antibodies/pgam5-antibody-rm1254-ab322207'>ab322207</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Western blot - Human PGAM5 knockout HeLa cell lysate (<a href='/en-us/products/cell-lysates/human-pgam5-knockout-hela-cell-lysate-ab257581'>ab257581</a>) at 20 µg

Lane 3:

A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 32 kDa,36 kDa

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