Anti-Tet2 antibody

• WB

AbReview37584****

Western blot - Anti-Tet2 antibody (AB124297)

All lanes:

Western blot - Anti-Tet2 antibody (ab124297) at 1/500 dilution

Lane 1:

Wild-type mouse embryonic stem cells at 20 µg

Lane 2:

Tet2-/- mouse embryonic stem cells at 20 µg

Secondary

All lanes:

Goat anti-Rabbit (whole molecules) IgG HRP at 1/3000 dilution

Predicted band size: 223 kDa

Observed band size: 250 kDa

true

Exposure time: 2min

This image is courtesy of an anonymous abreview.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Tet2 antibody (AB124297)

ab124297 staining Tet2 in mES cells (Positive cell line) and NIH/3T3 (Negative cell line). The cells were fixed with 4% paraformaldehyde (10 min) permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab124297 at 1µg/ml and ab7291 Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150081 Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488) pre-adsorbed at 1/1000 dilution (shown in green) and ab150120  Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594) pre-adsorbed at 1/1000 dilution (shown in pseudocolour magenta). Nuclear DNA was labelled with DAPI (shown in blue).

Image was acquired with a confocal microscope (Leica-Microsystems TCS SP8) and a single confocal section is shown.

• WB

Lab

Western blot - Anti-Tet2 antibody (AB124297)

Western blot : Anti-TET2 antibody (ab124297) staining at 1/250 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab124297 was shown to bind specifically to TET2. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Tet2 antibody (ab124297) at 1/250 dilution

Lane 1:

mES cell lysate at 20 µg

Lane 2:

F9 cell lysate at 20 µg

Lane 3:

NIH/3T3 cell lysate at 20 µg

Observed band size: 225 kDa

false

• WB

AbReview51133****

Western blot - Anti-Tet2 antibody (AB124297)

All lanes:

Western blot - Anti-Tet2 antibody (ab124297) at 1/1000 dilution

Lane 1:

Wild type mouse embryonic stem cells at 20 µg with Milk, 1 hour at 21°C

Lane 2:

Tet2 KO clone#1 mouse embryonic stem cells at 20 µg with Milk, 1 hour at 21°C

Lane 3:

Tet2 KO clone#2 mouse embryonic stem cells at 20 µg with Milk, 1 hour at 21°C

Secondary

All lanes:

HRP conjugated Goat anti-rabbit IgG at 1/10000 dilution

Predicted band size: 223 kDa

true

Exposure time: 5min

This image is courtesy of an anonymous abreview

• ICC/IF

Collaborator

Immunocytochemistry/ Immunofluorescence - Anti-Tet2 antibody (AB124297)

ICC/IF image of Anti-Tet2 antibody (ab124297) stained WT and Tet2^-/- mouse ES cells. The cells were fixed in PFA, permeabilized using 0.25% Triton X-100, and blocked with 10% BSA for 30 minutes. The cells were then incubated with ab124297 at a 1/400 dilution for 13 hours and 20 minutes at 4^oC. The secondary antibody was a Rhodamine Red-X AffiniPure Donkey anti-Rabbit used at a 1/500 dilution.

This image is courtesy of an anonymous abreview.

• ICC/IF

Collaborator

Immunocytochemistry/ Immunofluorescence - Anti-Tet2 antibody (AB124297)

ICC/IF image of Anti-Tet2 antibody (ab124297) stained D3 mouse ES cells. The cells were fixed in 4% PFA, permeabilized using 0.1% Triton X-100, and blocked with 1% Goat serum, 0.1% BSA in PBS for 30 minutes. The cells were then incubated with ab124297 at a 1/100 dilution for 2 hours at RT. The secondary antibody was a Goat polyclonal Secondary Antibody to Rabbit IgG – H&L Alexa Fluor 488 (ab150077) used at a 1/200 dilution.

Secondary antibody - goat anti-rabbit Alexa 488 (ab150077)

This image is courtesy of an abreview from Joe Segal.

• WB

Unknown

Western blot - Anti-Tet2 antibody (AB124297)

This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 1% Milk before being incubated with ab124297 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

Secondary antibody - goat anti-rabbit HRP (ab97051)

All lanes:

Western blot - Anti-Tet2 antibody (ab124297) at 1/250 dilution

Lane 1:

E14Tg2a (Mouse embryonic stem cell line) Whole Cell Lysate at 25 µg

Lane 2:

WT Mouse ES Cell Lysate (Positive Control for Tet2) at 25 µg

Lane 3:

Tet2 Knockout Mouse ES Cell Lysate (Negative Control) at 25 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 1/10000 dilution

Predicted band size: 223 kDa

Observed band size: 223 kDa,48 kDa,65 kDa,70 kDa

true

Exposure time: 4min