Anti-IP6K1 (phospho S118 + S121) antibody [EPR19852]

• IP

Unknown

Immunoprecipitation - Anti-IP6K1 (phospho S118 + S121) antibody [EPR19852] (AB214816)

IP6K1 (phospho S118 + S121) was immunoprecipitated from 0.35 mg MOLT-4 (human lymphoblastic leukemia cell line) whole cell lysate with ab214816 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab214816 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5,000 dilution

Lane 1 : MOLT-4 whole cell lysate 10 μg (Input).
Lane 2 : ab214816 IP in MOLT-4 whole cell lysate (+).
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab214816 in MOLT-4 whole cell lysate (-).

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

Exposure time : 1 minute.

All lanes:

Immunoprecipitation - Anti-IP6K1 (phospho S118 + S121) antibody [EPR19852] (ab214816)

Predicted band size: 50 kDa

Observed band size: 52 kDa

false

• WB

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Western blot - Anti-IP6K1 (phospho S118 + S121) antibody [EPR19852] (AB214816)

Blocking/diluting buffer : 5% NFDM/TBST.

The bands migrating at approximately 100 kDa are likely to be non-specific.

All lanes:

Western blot - Anti-IP6K1 (phospho S118 + S121) antibody [EPR19852] (ab214816) at 1/5000 dilution

Lane 1:

MOLT-4 (human lymphoblastic leukemia cell line) whole cell lysate at 10 µg

Lane 2:

MOLT-4 whole cell lysate. The WB membrane was treated with alkaline phosphatase for 1h at 37°C at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 50 kDa

Observed band size: 52 kDa

false

Exposure time: 26s

• WB

Unknown

Western blot - Anti-IP6K1 (phospho S118 + S121) antibody [EPR19852] (AB214816)

Blocking/diluting buffer : 5% NFDM/TBST.

The band at approximately 30 kDa likely represents degradation fragments of IP6K1 (pS118 and pS121) protein.

The plasmids were kindly provided by our collaborator Dr. Feng Rao NIBS.

All lanes:

Western blot - Anti-IP6K1 (phospho S118 + S121) antibody [EPR19852] (ab214816) at 1/10000 dilution

Lane 1:

HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) transfected with a His-tagged human IP6K1 (WT) expression vector, whole cell lysate at 10 µg

Lane 2:

HEK-293T transfected with a His-tagged human IP6K1 (WT) expression vector whole cell lysate. The WB membrane was treated with alkaline phosphatase for 1h at 37°C at 10 µg

Lane 3:

HEK-293T transfected with a His-tagged human IP6K1 S118A+S121A mutant expression vector, whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/ko/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 50 kDa

false

Exposure time: 26s

• Dot

Unknown

Dot Blot - Anti-IP6K1 (phospho S118 + S121) antibody [EPR19852] (AB214816)

Dot blot analysis of IP6K1 (phospho S118 + S121) labeled with ab214816 at 1/1000 dilution. Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody was used at 1/100,000 dilution..

Lane 1 : IP6K1 (phospho S118 + S121) peptide (aa114-125).

Lane 2 : IP6K1 (phospho S118 + S121) peptide (aa116-127).

Lane 3 : IP6K1 (phospho S118) peptide (aa114-127).

Lane 4 : IP6K1 (phospho S121) peptide (aa114-127).

Lane 5 : IP6K1 non-phospho peptide (aa114-127).

Exposure time : 58 seconds.

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