Anti-MTF2 antibody [EPR23536-87] - ChIP Grade - BSA and Azide free

• IP

Unknown

Immunoprecipitation - Anti-MTF2 antibody [EPR23536-87] - ChIP Grade - BSA and Azide free (AB275967)

This data was developed using ab254336, the same antibody clone in a different buffer formulation.

MTF2 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab254336 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254336 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 ug

Lane 2 : ab254336 IP in HeLa whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab254336 in HeLa whole cell lysate

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

Exposure time : 24 seconds.

67, 56, 47, 29-kDa bands correspond to MTF2 isoforms.This blot was developed using a higher sensitivity ECL substrate.

All lanes:

Immunoprecipitation - Anti-MTF2 antibody [EPR23536-87] - ChIP Grade (<a href='/en-us/products/primary-antibodies/mtf2-antibody-epr23536-87-chip-grade-ab254336'>ab254336</a>)

Predicted band size: 67 kDa

Observed band size: 29 kDa,47 kDa,56 kDa,67 kDa

false

• IP

Unknown

Immunoprecipitation - Anti-MTF2 antibody [EPR23536-87] - ChIP Grade - BSA and Azide free (AB275967)

This data was developed using ab254336, the same antibody clone in a different buffer formulation.

MTF2 was immunoprecipitated from 0.35 mg ES-D3 (mouse embryonic multipotent stem Cell) whole cell lysate with ab254336 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab254336 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : ES-D3 (mouse embryonic multipotent stem Cell) whole cell lysate 10 ug

Lane 2 : ab254336 IP in ES-D3 whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab254336 in ES-D3 whole cell lysate

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

Exposure time : 8 seconds.

67, 56, 47, 29-kDa bands correspond to MTF2 isoforms. Bands between 30-37 kDa is caused by degradation as demonstrated by WB data.

All lanes:

Immunoprecipitation - Anti-MTF2 antibody [EPR23536-87] - ChIP Grade (<a href='/en-us/products/primary-antibodies/mtf2-antibody-epr23536-87-chip-grade-ab254336'>ab254336</a>)

Predicted band size: 67 kDa

Observed band size: 29 kDa,47 kDa,56 kDa,67 kDa

false

• ChIP

Supplier Data

ChIP - Anti-MTF2 antibody [EPR23536-87] - ChIP Grade - BSA and Azide free (AB275967)

This data was developed using ab254336, the same antibody clone in a different buffer formulation.

Chromatin was prepared from ES-D3 cells according to the Abcam Dual-X-ChIP protocol. Cells were fixed with 1.5 mM EGS for 30mins and then formaldehyde for 10min.
The ChIP was performed with 25 μg of chromatin, 5 μg of ab254336 (red), or 5 μg of rabbit normal IgG ab172730 (gray) and 25 μl of Protein A/G Dynabeads. The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Primers are from paper PMID : 20123894

• WB

Lab

Western blot - Anti-MTF2 antibody [EPR23536-87] - ChIP Grade - BSA and Azide free (AB275967)

This data was developed using ab254336, the same antibody clone in a different buffer formulation.

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

The molecular weight observed is consistent with what has been described in the literature (PMID : 28869966).

Lysate loaded onto lane 6 was made freshly and used in WB immediately to minimize protein degradation.

67, 56, 47, 29-kDa bands correspond to MTF2 isoforms. Bands between 30-37 kDa in lane3 is caused by degradation.

Exposure time : Lanes 1-2 : 26 seconds; Lane 3 : 10 seconds; Lanes 4-5 : 125 seconds; Lane 6 : 3 minutes.

All lanes:

Western blot - Anti-MTF2 antibody [EPR23536-87] - ChIP Grade (<a href='/en-us/products/primary-antibodies/mtf2-antibody-epr23536-87-chip-grade-ab254336'>ab254336</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lanes 3 and 6:

ES-D3 (mouse embryonic mtipotent stem Cell), whole cell lysate 20 at 20 µg

Lane 4:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 5:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 67 kDa

Observed band size: 47 kDa,56 kDa,67 kDa

false

• WB

Lab

Western blot - Anti-MTF2 antibody [EPR23536-87] - ChIP Grade - BSA and Azide free (AB275967)

This data was developed using ab254336, the same antibody clone in a different buffer formulation.

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

Exposure time : 3 minutes.

All lanes:

Western blot - Anti-MTF2 antibody [EPR23536-87] - ChIP Grade (<a href='/en-us/products/primary-antibodies/mtf2-antibody-epr23536-87-chip-grade-ab254336'>ab254336</a>) at 1/1000 dilution

All lanes:

Human testis tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Predicted band size: 67 kDa

Observed band size: 47 kDa,56 kDa,67 kDa

false

• WB

Lab

Western blot - Anti-MTF2 antibody [EPR23536-87] - ChIP Grade - BSA and Azide free (AB275967)

This data was developed using ab254336, the same antibody clone in a different buffer formulation.

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

Exposure time : 92 seconds.

All lanes:

Western blot - Anti-MTF2 antibody [EPR23536-87] - ChIP Grade (<a href='/en-us/products/primary-antibodies/mtf2-antibody-epr23536-87-chip-grade-ab254336'>ab254336</a>) at 1/1000 dilution

Lane 1:

Mouse skeletal muscle tissue lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Lane 3:

Rat testis tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 67 kDa

Observed band size: 47 kDa,56 kDa,67 kDa

false