Anti-PMCA1 antibody [EPR12029] - BSA and Azide free

• IP

Lab

Immunoprecipitation - Anti-PMCA1 antibody [EPR12029] - BSA and Azide free (AB251018)

This data was developed using ab190355, the same antibody clone in a different buffer formulation.

Immunoprecipitation of ATP2B1 in HAP1 cells. Lysates were prepared and immunoprecipitation was performed using 2.0 µg of ab190355 pre-coupled to Protein A beads. Samples were then washed and processed for western blot.

These data were 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-PMCA1 antibody [EPR12029] (<a href='/en-us/products/primary-antibodies/pmca1-antibody-epr12029-ab190355'>ab190355</a>) at 2 µg

All lanes:

HAP1 cells

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMCA1 antibody [EPR12029] - BSA and Azide free (AB251018)

This data was developed using the same antibody clone in a different buffer formulation (ab190355).

Immunohistochemical analysis of paraffin-embedded mouse lung cancer tissue labelling PMCA1 with ab190355 at 1/2000 dilution, followed by ready-to-use secondary antibody Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at dilution 1/1. Counterstained with Hematoxylin.

Performed heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Positive staining mouse lung cancer.

The section was incubated with ab190355 for 10 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMCA1 antibody [EPR12029] - BSA and Azide free (AB251018)

This data was developed using the same antibody clone in a different buffer formulation (ab190355).

Immunohistochemical analysis of paraffin-embedded mouse stomach tissue labelling PMCA1 with ab190355 at 1/2000 dilution, followed by ready-to-use secondary antibody Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at dilution 1/1. Counterstained with Hematoxylin.

Performed heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Positive staining mouse stomach.

The section was incubated with ab190355 for 10 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-PMCA1 antibody [EPR12029] - BSA and Azide free (AB251018)

This data was developed using the same antibody clone in a different buffer formulation (ab190355).

Immunohistochemical analysis of paraffin-embedded rat stomach tissue labelling PMCA1 with ab190355 at 1/2000 dilution, followed by ready-to-use secondary antibody Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) at dilution 1/1. Counterstained with Hematoxylin.

Performed heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.

Positive staining mouse lung cancer.

The section was incubated with ab190355 for 10 mins at room temperature. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument.

• WB

Supplier Data

Western blot - Anti-PMCA1 antibody [EPR12029] - BSA and Azide free (AB251018)

This data was developed using the same antibody clone in a different buffer formulation (ab190355).

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

Exposure time :
Lanes 1, 2 : 7 seconds

Lanes 3, 4, 5, 6 : 3 seconds

ab181602 is used as a Loading Control at dilution 1/1000000.

All lanes:

Western blot - Anti-PMCA1 antibody [EPR12029] (<a href='/en-us/products/primary-antibodies/pmca1-antibody-epr12029-ab190355'>ab190355</a>) at 1/5000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate, boiled at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate, unboiled at 20 µg

Lane 3:

Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate, boiled at 20 µg

Lane 4:

Neuro-2a (Mouse neuroblastoma neuroblast) whole cell lysate, unboiled at 20 µg

Lane 5:

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

Lane 6:

C6 (Rat glial tumor glial cell) whole cell lysate, unboiled 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: 139 kDa

Observed band size: 130-250 kDa

false

• WB

Lab

Western blot - Anti-PMCA1 antibody [EPR12029] - BSA and Azide free (AB251018)

This data was developed using ab190355, the same antibody clone in a different buffer formulation.

ab190355 was shown to react with ATP2B1 in wild-type HAP1 cells in Western blot with loss of signal observed in a ATP2B1 knockout cell line. Wild-type HAP1 and ATP2B1 knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 5% milk in TBST for 1 hr before incubation with ab190355 overnight at 4 °C at a 1/1000 dilution. Blots were incubated with secondary antibodies at 0.2 µg/mL before imaging.

These data were 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-PMCA1 antibody [EPR12029] (<a href='/en-us/products/primary-antibodies/pmca1-antibody-epr12029-ab190355'>ab190355</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 lysate at 30 µg

Lane 2:

ATP2B1 knock-out HAP1 lysate at 30 µg

false

• WB

Lab

Western blot - Anti-PMCA1 antibody [EPR12029] - BSA and Azide free (AB251018)

This data was developed using the same antibody clone in a different buffer formulation (ab190355).

Lanes 1-4 : Merged signal (red and green). Green - ab190355 observed at 180-245 kDa. Red - loading control ab8245 observed at 36 kDa.

ab190355 Anti-PMCA1 antibody [EPR12029] was shown to specifically react with PMCA1 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265561 (knockout cell lysate ab257365) was used. Wild-type and PMCA1 knockout samples were subjected to SDS-PAGE. ab190355 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2. 5 hours 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-PMCA1 antibody [EPR12029] (<a href='/en-us/products/primary-antibodies/pmca1-antibody-epr12029-ab190355'>ab190355</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

ATP2B1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human ATP2B1 (PMCA1) knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-atp2b1-pmca1-knockout-hela-cell-line-ab265561'>ab265561</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Lane 4:

THP-1 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: 139 kDa

Observed band size: 180-245 kDa

false