重组Anti-PGAM5 (Long isoform)抗体[EPR27145-21]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGAM5 (Long isoform) antibody [EPR27145-21] (AB308447)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labelling PGAM5 (Long isoform) with ab308447 at 1/100 (4.625 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human colon. The section was incubated with ab308447 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGAM5 (Long isoform) antibody [EPR27145-21] (AB308447)

Immunohistochemical analysis of paraffin-embedded Human ovarian carcin tissue labelling PGAM5 (Long isoform) with ab308447 at 1/100 (4.625 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human ovarian carcinoma. The section was incubated with ab308447 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGAM5 (Long isoform) antibody [EPR27145-21] (AB308447)

Immunohistochemical analysis of paraffin-embedded (A) Parental Hela ( tissue labelling PGAM5 (Long isoform) with ab308447 at 1/2000 (0.231 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on (A) parental Hela cell pellet, no staining on (B) PGAM5 knockout Hela cell pellet. The section was incubated with ab308447 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGAM5 (Long isoform) antibody [EPR27145-21] (AB308447)

Immunohistochemical analysis of paraffin-embedded Rat colon tissue labelling PGAM5 (Long isoform) with ab308447 at 1/100 (4.625 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat colon. The section was incubated with ab308447 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PGAM5 (Long isoform) antibody [EPR27145-21] (AB308447)

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labelling PGAM5 (Long isoform) with ab308447 at 1/100 (4.625 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse testis. The section was incubated with ab308447 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

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Western blot - Anti-PGAM5 (Long isoform) antibody [EPR27145-21] (AB308447)

Blocking and diluting buffer and concentration : Intercept® (TBS) Blocking Buffer diluted with an equal volume of TBS. Lysates at 20 µg per lane. The samples were run on a Bis-Tris gel under reducing conditions. Western blot : Anti-PGAM5 (Long isoform) antibody (ab308447) staining at 1/1000 dilution, shown in green; Mouse anti-Tubulin antibody DM1A (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab308447 was shown to bind specifically to PGAM5 (Long isoform).  Target of interest was observed at 32 kDa and 36 KDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in PGAM5 knockout cell line ab265141 (knockout cell lysate ab257581) (lane 2). To generate this image, samples were first run on an SDS-PAGE gel then transferred onto an immobilon-FL PVDF membrane. Membranes were blocked in a fluorescentwestern blot (TBS-based) blocking solution before incubation with primaryantibodies overnight at 4 °C.  Blots were washed four times in TBS-T, incubated with secondary antibodies Goatanti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution for 1 h at room temperature, washed again then imaged.

All lanes:

Western blot - Anti-PGAM5 (Long isoform) antibody [EPR27145-21] (ab308447) at 1/1000 dilution

Lane 1:

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

Lane 2:

PGAM5 knockout HeLa whole cell lysate at 20 µg

Lane 3:

MEF (mouse embryonic fibroblast (immortalized)) whole cell lysate at 20 µg

Lane 4:

C6 (rat glial tumor glial cell) whole cell lysate, at 20 µg

Predicted band size: 32 kDa

Observed band size: 32 kDa

false

• WB

Supplier Data

Western blot - Anti-PGAM5 (Long isoform) antibody [EPR27145-21] (AB308447)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 26 seconds

All lanes:

Western blot - Anti-PGAM5 (Long isoform) antibody [EPR27145-21] (ab308447) at 1/1000 dilution

Lane 1:

Untreated PANC-1(human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

PANC-1 treated with 100 uM CCCP (141229) for 2 hours, whole cell lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 32 kDa

Observed band size: 32 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Anti-PGAM5 (Long isoform) antibody [EPR27145-21] (AB308447)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Lanes 2-3 were developed using a high sensitivity ECL substrate. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-PGAM5 (Long isoform) antibody [EPR27145-21] (ab308447) at 1/1000 dilution

Lane 1:

Human liver tissue lysate at 20 µg

Lane 2:

Mouse colon tissue lysate, at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 32 kDa

Observed band size: 32 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-PGAM5 (Long isoform) antibody [EPR27145-21] (AB308447)

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PGAM5 (Long isoform) antibody [EPR27145-21] (ab308447) at 1/1000 dilution

Lane 1:

Rat colon tissue lysate at 20 µg

Lane 2:

Rat liver tissue lysate at 20 µg

Lane 3:

Rat kidney tissue lysate at 20 µg

Lane 4:

Rat brain tissue lysate at 20 µg

Secondary

All lanes:

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

Predicted band size: 32 kDa

Observed band size: 32 kDa

true

Exposure time: 180s