Anti-Zfp57 antibody [EPR28485-527] - BSA and Azide free

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Zfp57 antibody [EPR28485-527] - BSA and Azide free (AB324146)

This data was developed using ab324145, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse testis tissue labeling Zfp57 with ab324145 at 1/50 (10.04 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on mouse testis (PMID : 8120052). The section was incubated with ab324145 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Zfp57 antibody [EPR28485-527] - BSA and Azide free (AB324146)

This data was developed using ab324145, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse pancreas carcinoma tissue labeling Zfp57 with ab324145 at 1/50 (10.04 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Positive staining on mouse pancreas carcinoma. The section was incubated with 324145 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ChIP-seq

Supplier Data

ChIP-sequencing - Anti-Zfp57 antibody [EPR28485-527] - BSA and Azide free (AB324146)

This data was developed using ab324145, the same antibody clone in a different buffer formulation.

Chromatin was prepared from ES-D3 [D3] cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5x10⁶ cells and 4 µg of ab324145 [EPR28485-527]. 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-Zfp57 antibody [EPR28485-527] - BSA and Azide free (AB324146)

This data was developed using ab324145, the same antibody clone in a different buffer formulation.

Chromatin was prepared from ES-D3 [D3] cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5x10⁶ cells and 4 µg of ab324145 [EPR28485-527]. 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-Zfp57 antibody [EPR28485-527] - BSA and Azide free (AB324146)

This data was developed using ab324145, the same antibody clone in a different buffer formulation.

Chromatin was prepared from ES-D3 [D3] cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5x10⁶ cells and 4 µg of ab324145 [EPR28485-527]. 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-Zfp57 antibody [EPR28485-527] - BSA and Azide free (AB324146)

This data was developed using ab324145, the same antibody clone in a different buffer formulation.

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

Low expression : A20.

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.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Zfp57 antibody [EPR28485-527] (<a href='/en-us/products/primary-antibodies/zfp57-antibody-epr28485-527-ab324145'>ab324145</a>) at 1/1000 dilution

Lanes 1 and 3:

ES-D3 [D3] (mouse blastocyst-derived embryonic stem cell) whole cell lysate at 20 µg

Lane 2:

A20 (mouse reticulum sarcoma b lymphocyte) 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 1/20000 dilution

Observed band size: 48 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Zfp57 antibody [EPR28485-527] - BSA and Azide free (AB324146)

This data was developed using ab324145, the same antibody clone in a different buffer formulation.

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

Low expression : skeletal muscle(PMID : 15070898).

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.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Zfp57 antibody [EPR28485-527] (<a href='/en-us/products/primary-antibodies/zfp57-antibody-epr28485-527-ab324145'>ab324145</a>) at 1/1000 dilution

Lane 1:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Mouse testis tissue lysate at 20 µg

Lane 3:

Mouse skeletal muscle 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 1/20000 dilution

Observed band size: 48 kDa,36 kDa

false

Exposure time: 180s

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Zfp57 antibody [EPR28485-527] - BSA and Azide free (AB324146)

This data was developed using ab324145, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling Zfp57 with ab324145 at 1/50 (10.04 ug/ml) dilution, followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Negative control : no staining on mouse skeletal muscle. The section was incubated with ab324145 for 30 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument

Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• Dot

Supplier Data

Dot Blot - Anti-Zfp57 antibody [EPR28485-527] - BSA and Azide free (AB324146)

This data was developed using ab324145, the same antibody clone in a different buffer formulation.

Dot blot analysis of Zfp57 using ab324145 at 1 : 1000 (0.502 ug/ml) followed by a Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1 : 100,000 dilution.

Lane1 : Mouse Zfp57 peptide
Lane2 : Mouse Znf182 peptide

Exposure time : 180 seconds.

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

This antibody does not cross-react with mouse Znf182 peptide by dot blot.

All lanes:

Dot Blot - Anti-Zfp57 antibody [EPR28485-527] (<a href='/en-us/products/primary-antibodies/zfp57-antibody-epr28485-527-ab324145'>ab324145</a>) at 1/1000 dilution

Lane 1:

Mouse Zfp57 peptide

Lane 2:

Mouse Znf182 peptide

Secondary

All lanes:

Dot 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

false

Exposure time: 180s