Anti-Peroxiredoxin 1/PAG antibody [EPR5433] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Peroxiredoxin 1/PAG antibody [EPR5433] - BSA and Azide free (AB247879)

This data was developed using the same antibody clone in a different buffer formulation (ab109498).

Immunohistochemical analysis of paraffin-embedded Human thyroid cancer tissue labeling Peroxiredoxin 1/PAG with ab109498 at 1/1000 (0.155 ug/ml) dilution. The section was incubated with ab109498 for 30 mins at room temperature and followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Counterstained with Hematoxylin.

Positive staining on the human thyroid cancer, performed on a Leica Biosystems BOND® RX instrument.

Secondary antibody only control : Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Peroxiredoxin 1/PAG antibody [EPR5433] - BSA and Azide free (AB247879)

This data was developed using the same antibody clone in a different buffer formulation (ab109498).

Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Peroxiredoxin 1/PAG with ab109498 at 1/4000 (0.038 ug/ml) dilution. The section was incubated with ab109498 for 30 mins at room temperature and followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Counterstained with Hematoxylin.

Positive staining on the mouse kidney, performed on a Leica Biosystems BOND® RX instrument.

Secondary antibody only control : Rabbit specific IHC polymer detection kit HRP/DAB (ab209101)

Heat mediated antigen retrieval with Citrate buffer (pH 6.0, epitope retrieval solution 1) for 20 mins.

• WB

Supplier Data

Western blot - Anti-Peroxiredoxin 1/PAG antibody [EPR5433] - BSA and Azide free (AB247879)

This data was developed using the same antibody clone in a different buffer formulation (ab109498).

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

All lanes:

Western blot - Anti-Peroxiredoxin 1/PAG antibody [EPR5433] (<a href='/en-us/products/primary-antibodies/peroxiredoxin-1-pag-antibody-epr5433-ab109498'>ab109498</a>) at 1/10000 dilution

Lane 1:

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

Lane 2:

K-562 (Human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 3:

NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

Mouse brain 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 1/20000 dilution

Predicted band size: 22 kDa

Observed band size: 22 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-Peroxiredoxin 1/PAG antibody [EPR5433] - BSA and Azide free (AB247879)

This data was developed using the same antibody clone in a different buffer formulation (ab109498).

Lane 1 : Wild-type HAP1 cell lysate (20 μg)
Lane 2 : Peroxiredoxin 1/PAG knockout HAP1 cell lysate (20 μg)
Lane 3 : A431 cell lysate (20 μg)
Lane 4 : Jurkat cell lysate (20 μg)
Lanes 1 - 4 : Merged signal (red and green). Green - ab109498 observed at 23 kDa. Red - loading control, ab8245, observed at 37 kDa.

ab109498 was shown to specifically react with Peroxiredoxin 1/PAG when Peroxiredoxin 1/PAG knockout samples were used. Wild-type and Peroxiredoxin 1/PAG knockout samples were subjected to SDS-PAGE. ab109498 and ab8245 (loading control to GAPDH) were both diluted 1/10 000 and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10 000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-Peroxiredoxin 1/PAG antibody [EPR5433] - BSA and Azide free (ab247879)

Predicted band size: 22 kDa

false

• WB

Lab

Western blot - Anti-Peroxiredoxin 1/PAG antibody [EPR5433] - BSA and Azide free (AB247879)

This data was developed using the same antibody clone in a different buffer formulation (ab109498).

Lanes 1-3 : Merged signal (red and green). Green - ab109498 observed at 26 kDa. Red - loading control ab8245 observed at 36 kDa.

ab109498 Anti-Peroxiredoxin 1/PAG antibody [EPR5433] was shown to specifically react with Peroxiredoxin in wild-type HEK293T cells. Loss of signal was observed when knockout cell line ab266842 (knockout cell lysate ab257040) was used. Wild-type and Peroxiredoxin knockout samples were subjected to SDS-PAGE. ab109498 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-Peroxiredoxin 1/PAG antibody [EPR5433] (<a href='/en-us/products/primary-antibodies/peroxiredoxin-1-pag-antibody-epr5433-ab109498'>ab109498</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T cell lysate at 20 µg

Lane 2:

PRDX1 knockout HEK293T cell lysate at 20 µg

Lane 2:

Western blot - Human PRDX1 (Peroxiredoxin 1/PAG) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-prdx1-peroxiredoxin-1-pag-knockout-hek-293t-cell-line-ab266842'>ab266842</a>)

Lane 3:

Jurkat cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 22 kDa,41 kDa,45 kDa,78 kDa

Observed band size: 25 kDa,26 kDa,47 kDa,49 kDa,78 kDa

false