Anti-Tet2 antibody [EPR29630-29] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-Tet2 antibody [EPR29630-29] - BSA and Azide free (AB328046)

This data was developed using ab324875, the same antibody clone in a different buffer formulation.

Tet2 was immunoprecipitated from 0.35 mg 293T (human embryonic kidney epithelial cell) whole cell fresh lysate with ab324875 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab324875 at 1/1000 dilution.

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

The bands beneath the target band (260 kDa) are likely to be degraded target fragments.
To minimize protein degradation cells were lysed immediately after harvest and then applied for Immunoprecipitation as soon as possible.

All lanes:

Immunoprecipitation - Anti-Tet2 antibody [EPR29630-29] (<a href='/products/primary-antibodies/tet2-antibody-epr29630-29-ab324875'>ab324875</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) whole cell fresh lysate at 10 µg

Lane 2:

<a href='/products/primary-antibodies/tet2-antibody-epr29630-29-ab324875'>ab324875</a> at 1/30 IP in 293T (human embryonic kidney epithelial cell) whole cell fresh lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/products/primary-antibodies/tet2-antibody-epr29630-29-ab324875'>ab324875</a> in 293T whole cell fresh lysate

Secondary

All lanes:

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

Observed band size: 260 kDa

false

Exposure time: 180s

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-Tet2 antibody [EPR29630-29] - BSA and Azide free (AB328046)

This data was developed using ab324875, the same antibody clone in a different buffer formulation.

Chromatin was prepared from 293T cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 µg of ab324875 [EPR29630-29]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-Tet2 antibody [EPR29630-29] - BSA and Azide free (AB328046)

This data was developed using ab324875, the same antibody clone in a different buffer formulation.

Chromatin was prepared from 293T cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 µg of ab324875 [EPR29630-29]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-Tet2 antibody [EPR29630-29] - BSA and Azide free (AB328046)

This data was developed using ab324875, the same antibody clone in a different buffer formulation.

Chromatin was prepared from 293T cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 10⁷ cells and 8 µg of ab324875 [EPR29630-29]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• WB

Supplier Data

Western blot - Anti-Tet2 antibody [EPR29630-29] - BSA and Azide free (AB328046)

This data was developed using ab324875, the same antibody clone in a different buffer formulation.

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

Performed under reducing conditions.

In Western blot ab324875 was shown to bind specifically to Tet2 . Target of interest was observed at 260 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in Tet2 knockout cell line (lane 2).

To minimize protein degradation cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The bands beneath the target band (260 kDa) are likely to be degraded target fragments.

In Western blot Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-Tet2 antibody [EPR29630-29] (<a href='/products/primary-antibodies/tet2-antibody-epr29630-29-ab324875'>ab324875</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 2:

Tet2 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

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

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 260 kDa,124 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Tet2 antibody [EPR29630-29] - BSA and Azide free (AB328046)

This data was developed using ab324875, the same antibody clone in a different buffer formulation.

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

This blot was developed using a high-sensitivity ECL substrate allowing for the detection of proteins in the mid-femtogram range.

To minimize protein degradation cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The bands beneath the target band (260 kDa) are likely to be degraded target fragments.

In Western blot Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-Tet2 antibody [EPR29630-29] (<a href='/products/primary-antibodies/tet2-antibody-epr29630-29-ab324875'>ab324875</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with scrambled siRNA control whole cell lysate at 10 µg

Lane 2:

293T transfected with siRNA specifically targeting Tet2 whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 260 kDa,124 kDa

true

Exposure time: 180s