Anti-SHP2 antibody [EPR17829-9]

• IP

Supplier Data

Immunoprecipitation - Anti-SHP2 antibody [EPR17829-9] (AB187040)

SHP2 was immunoprecipitated from 1 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab187040 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab187040 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

Lane 1 : HeLa whole cell lysate 10 μg (Input).

Lane 2 : ab187040 IP in HeLa whole cell lysate (+).

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab187040 in HeLa whole cell lysate (-).

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

Exposure time : 3 minutes.

All lanes:

Immunoprecipitation - Anti-SHP2 antibody [EPR17829-9] (ab187040)

Predicted band size: 43 kDa,68 kDa

Observed band size: 68 kDa

true

• IP

Supplier Data

Immunoprecipitation - Anti-SHP2 antibody [EPR17829-9] (AB187040)

SHP2 was immunoprecipitated from 1 mg of NIH/3T3 (mouse embryo fibroblast cell line) whole cell lysate with ab187040 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab187040 at 1/1,000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10,000 dilution.

Lane 1 : NIH/3T3 whole cell lysate 10μg (Input).

Lane 2 : ab187040 IP in NIH/3T3 whole cell lysate (+).

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab187040 in NIH/3T3 whole cell lysate (-).

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

Exposure time : 1 second.

All lanes:

Immunoprecipitation - Anti-SHP2 antibody [EPR17829-9] (ab187040)

Predicted band size: 68 kDa

Observed band size: 68 kDa

true

• WB

Supplier Data

Western blot - Anti-SHP2 antibody [EPR17829-9] (AB187040)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-SHP2 antibody [EPR17829-9] (ab187040) at 1/5000 dilution

Lane 1:

HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 5 µg

Lane 2:

Jurkat (human T cell leukemia cell line from peripheral blood) whole cell lysate at 5 µg

Lane 3:

HEK-293 (human epithelial cell line from embryonic kidney) whole cell lysate at 5 µg

Lane 4:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 5 µg

Lane 5:

Mouse brain tissue lysate at 5 µg

Lane 6:

Mouse heart tissue lysate at 5 µg

Lane 7:

Rat brain tissue lysate at 5 µg

Lane 8:

Rat heart tissue lysate at 5 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

Predicted band size: 68 kDa

Observed band size: 68 kDa

true

Exposure time: 1min

• WB

Lab

Western blot - Anti-SHP2 antibody [EPR17829-9] (AB187040)

Lanes 1- 2 : Merged signal (red and green). Green - ab187040 observed at 68 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab187040 was shown to react with SHP2 in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab266450 (knockout cell lysate ab257618) was used. Wild-type HEK-293T and PTPN11 knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab187040 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 5000 dilution and a 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-SHP2 antibody [EPR17829-9] (ab187040) at 1/5000 dilution

Lane 1:

Wild-type HEK-293T cell lysate at 20 µg

Lane 2:

PTPN11 knockout HEK-293T cell lysate at 20 µg

Lane 2:

Western blot - Human PTPN11 (SHP2) knockout HEK-293T cell line (<a href='/en-us/products/cell-lines/human-ptpn11-shp2-knockout-hek-293t-cell-line-ab266450'>ab266450</a>)

Predicted band size: 68 kDa

Observed band size: 68 kDa

false

• WB

Lab

Western blot - Anti-SHP2 antibody [EPR17829-9] (AB187040)

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

Exposure time Lane 1-8 : 180s

Exposure time Lane 9-10 : 1s

Exposure time Lane 11-16 : 15s

ab300579 is less sensitive than ab187040 in WB. We suggest customers choosing ab187040 for WB test.

Lanes 1 - 8:

Western blot - Anti-SHP2 antibody [EPR26539-45] (<a href='/en-us/products/primary-antibodies/shp2-antibody-epr26539-45-ab300579'>ab300579</a>) at 1/1000 dilution

Lanes 9 - 16:

Western blot - Anti-SHP2 antibody [EPR17829-9] (ab187040) at 1/1000 dilution

Lanes 1 and 9:

Human cerebellum tissue lysate, 20 μg

Lanes 2 and 10:

Human heart tissue lysate, 20 μg

Lanes 3 and 11:

Mouse ovary tissue lysate, 20 μg

Lanes 4 and 12:

Mouse embryo tissue lysate, 20 μg

Lanes 5 and 13:

Mouse brain tissue lysate, 20 μg

Lanes 6 and 14:

Mouse heart tissue lysate, 20 μg

Lanes 7 and 14:

Rat brain tissue lysate, 20 μg

Lanes 8 and 16:

Rat heart tissue lysate, 20 μg

Secondary

Lanes 1, 2, 9 and 10:

Goat Anti-Rabbit IgG (HRP), minimal cross-reactivity with human IgG at 1/2000 dilution

Lanes 3, 4, 5, 6, 7, 8, 11, 12, 13, 14, 15 and 16:

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

Predicted band size: 68 kDa

Observed band size: 72 kDa

false