重组Anti-Pyruvate Dehydrogenase E2抗体[EPR12455(B)]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (AB172617)

Immunohistochemical analysis of paraffin-embedded human cervical cancer labeling Pyruvate Dehydrogenase E2 with ab172617 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on human cervical cancer.
The section was incubated with ab172617 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (AB172617)

Immunohistochemical analysis of paraffin-embedded human cerebrum labeling Pyruvate Dehydrogenase E2 with ab172617 at 1/1000 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on human cerebrum.
The section was incubated with ab172617 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (AB172617)

ab172617 staining Pyruvate Dehydrogenase E2 in A549 (positive) and A549-DLAT (negative) cells. The cells were fixed with 100% methanol (5 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated for 2 hours at room temperature with ab172617 at 1 ug/ml (shown in green) and ab7291, Anti-alpha Tubulin antibody [DM1A] - Loading Control (shown in pseudocolour magenta). Cells were then incubated with ab150081, Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) preadsorbed at 1/1000 dilution and ab150119, Goat Anti-Mouse IgG H&L (Alexa Fluor^® 647) preadsorbed at 1/1000 dilution for 1 hour at room temperature. Nuclear DNA was labeled with DAPI (shown in blue). Also suitable in cells fixed with 4% paraformaldehyde (10 min). Image was acquired with a high-content analyser (Operetta CLS, Revvity) and a maximum intensity projection of confocal sections is shown.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (AB172617)

Intracellular flow cytometric analysis of permeabilized Jurkat cells labeling Pyruvate Dehydrogenase E2 with ab172617 at 1/10 dilution (red), compared to a rabbit IgG negative control (green).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (AB172617)

Immunofluorescent analysis of HepG2 cells labeling Pyruvate Dehydrogenase E2 with ab172617 at 1/50 dilution.

• IP

Supplier Data

Immunoprecipitation - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (AB172617)

Immunoprecipitation of DLAT in A549 cells. Lysates were prepared and immunoprecipitation was performed using 2 μg of ab172617 pre-coupled to Protein A beads. Samples were then washed and processed for western blot.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Immunoprecipitation - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (ab172617) at 2 µg

All lanes:

A549 cells lysates

Observed band size: 68 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (AB172617)

Western blot analysis on immunoprecipitation pellet from HepG2 cell lysate immunoprecipitated using ab172617 at 1/10 dilution.

All lanes:

Immunoprecipitation - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (ab172617)

Predicted band size: 69 kDa

false

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (AB172617)

Immunohistochemical analysis of paraffin-embedded rat kidney labeling Pyruvate Dehydrogenase E2 with ab172617 at 1/500 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on rat kidney.
The section was incubated with ab172617 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (AB172617)

Immunohistochemical analysis of paraffin-embedded mouse kidney labeling Pyruvate Dehydrogenase E2 with ab172617 at 1/500 dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on mouse kidney.
The section was incubated with ab172617 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 Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

Supplier Data

Western blot - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (AB172617)

All lanes:

Western blot - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (ab172617) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

HepG2 cell lysate at 10 µg

Lane 3:

Jurkat cell lysate at 10 µg

Lane 4:

293T cell lysate at 10 µg

Predicted band size: 69 kDa

false

• WB

Supplier Data

Western blot - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (AB172617)

ab172617 was shown to react with DLAT in wild-type A549 cells in Western blot with loss of signal observed in DLAT knockout cell line ab300919. Wild-type A549 and DLAT knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab172617 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

This data was provided by YCharOS Inc., an open science company with the mission of characterizing commercially available antibody reagents for all human proteins. Abcam and YCharOS are working together to help address the reproducibility crisis by enabling the life science community to better evaluate commercially available antibodies.

All lanes:

Western blot - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (ab172617) at 1/1000 dilution

Lane 1:

Wild-type A549 lysate at 30 µg

Lane 2:

DLAT knock-out A549 lysate at 30 µg

Lane 2:

Western blot - Human DLAT knockout A549 cell line (<a href='/products/cell-lines/human-dlat-knockout-a549-cell-line-ab300919'>ab300919</a>)

Observed band size: 68 kDa

false

• WB

Lab

Western blot - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (AB172617)

All lanes:

Western blot - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (ab172617) at 1/1000 dilution

Lane 1:

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

Lane 2:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

Lane 4:

NIH/3T3 (mouse embryonic fibroblast) 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

Observed band size: 69 kDa

false

Exposure time: 48s

• WB

Lab

Western blot - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (AB172617)

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

All lanes:

Western blot - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (ab172617) at 1/1000 dilution

Lane 1:

Rat brain tissue lysate at 20 µg

Lane 2:

Rat kidney 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

Observed band size: 69 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (AB172617)

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

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

All lanes:

Western blot - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (ab172617) at 1/1000 dilution

Lane 1:

NIH/3T3 (mouse embryonic fibroblast) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

NIH/3T3 transfected with siRNA specifically targeting Dlat-1 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 1/20000 dilution

Observed band size: 69 kDa

false

Exposure time: 15s

• WB

Lab

Western blot - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (AB172617)

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

All lanes:

Western blot - Anti-Pyruvate Dehydrogenase E2 antibody [EPR12455(B)] (ab172617) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 20 µg

Lane 2:

Mouse kidney 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

Observed band size: 69 kDa

false

Exposure time: 26s