Anti-TRIP13/PCH2 antibody [EPR27063-80] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-TRIP13/PCH2 antibody [EPR27063-80] - BSA and Azide free (AB313703)

This data was developed using ab313702, the same antibody clone in a different buffer formulation. TRIP13/PCH2 was immunoprecipitated from 0.35 mg HEK-293 (human embryonic kidney epithelial cell) whole cell lysate with ab313702 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab313702 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution. Lane 1 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate Lane 2 : HEK-293 (human embryonic kidney epithelial cell) whole cell lysate Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab313702 in HEK-293 whole cell lysate Blocking and dilution buffer and concentration : 5% NFDM/TBST. Exposure time : 180 seconds

All lanes:

Immunoprecipitation - Anti-TRIP13/PCH2 antibody [EPR27063-80] (<a href='/en-us/products/primary-antibodies/trip13-pch2-antibody-epr27063-80-ab313702'>ab313702</a>) at 1/30 dilution

All lanes:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate

Secondary

All lanes:

Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (<a href='/en-us/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/5000 dilution

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-TRIP13/PCH2 antibody [EPR27063-80] - BSA and Azide free (AB313703)

This data was developed using ab313702, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. Exposure time : 180 seconds

All lanes:

Western blot - Anti-TRIP13/PCH2 antibody [EPR27063-80] (<a href='/en-us/products/primary-antibodies/trip13-pch2-antibody-epr27063-80-ab313702'>ab313702</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 3:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell 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 1/100000 dilution

Observed band size: 49 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-TRIP13/PCH2 antibody [EPR27063-80] - BSA and Azide free (AB313703)

This data was developed using ab313702, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : human skeletal muscle, human pancreas. Performed under reducing conditions. In Western blot, ab313702 was shown to bind specifically to TRIP13. Target of interest was observed at 49 kDa in wild-type HEK293T cell lysates (lane 1, wild-type cell line ab255449) with no signal observed at this size in TRIP13 knockout cell line (lane 2, knockout cell line ab266495 / knockout cell lysate ab258736). In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200000 dilution. Exposure time : 180 seconds

All lanes:

Western blot - Anti-TRIP13/PCH2 antibody [EPR27063-80] (<a href='/en-us/products/primary-antibodies/trip13-pch2-antibody-epr27063-80-ab313702'>ab313702</a>) at 1/1000 dilution

Lane 1:

Wild-type HEK293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

TRIP13 knockout HEK293T whole cell lysate at 20 µg

Lane 3:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 4:

Human testis tissue lysate at 20 µg

Lane 5:

Human skeletal muscle tissue lysate at 20 µg

Lane 6:

Human pancreas tissue lysate at 20 µg

Lane 7:

Rat testis tissue 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 1/100000 dilution

Observed band size: 49 kDa

false

Exposure time: 180s