Anti-n-Myc/MYCN antibody [EPR26107-153] - BSA and Azide free

• IP

Lab

Immunoprecipitation - Anti-n-Myc/MYCN antibody [EPR26107-153] - BSA and Azide free (AB325653)

This data was developed using ab325641, the same antibody clone in a different buffer formulation.

n-Myc/MYCN was immunoprecipitated from 0.35 mg IMR-32 (human neuroblastoma neuroblast) whole cell lysate with ab325641 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab325641 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : IMR-32 (human neuroblastoma neuroblast) whole cell lysate
Lane 2 : ab325641 IP in IMR-32 (human neuroblastoma neuroblast) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab325641 in IMR-32 whole cell lysate

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

Exposure time : 180 seconds.

All lanes:

Immunoprecipitation - Anti-n-Myc/MYCN antibody [EPR26107-153] (<a href='/en-us/products/primary-antibodies/n-myc-mycn-antibody-epr26107-153-ab325641'>ab325641</a>) at 1/1000 dilution

Lane 1:

IMR-32 (human neuroblastoma neuroblast) whole cell lysate at 10 µg

Lane 2:

<a href='/en-us/products/primary-antibodies/n-myc-mycn-antibody-epr26107-153-ab325641'>ab325641</a> at 1/30 IP in IMR-32 (human neuroblastoma neuroblast) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/n-myc-mycn-antibody-epr26107-153-ab325641'>ab325641</a> in IMR-32 whole cell lysate

Secondary

All lanes:

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

Observed band size: 60 kDa

false

Exposure time: 180s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-n-Myc/MYCN antibody [EPR26107-153] - BSA and Azide free (AB325653)

This data was developed using ab325641, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse E10.5 brain tissue labeling n-Myc/MYCN with ab325641 at 1/500 (1.042 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on brain of mouse E10.5. The primary antibody was incubated 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-n-Myc/MYCN antibody [EPR26107-153] - BSA and Azide free (AB325653)

This data was developed using ab325641, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse skin tissue labeling n-Myc/MYCN with ab325641 at 1/500 (1.042 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on hair follicle of mouse skin. The primary antibody was incubated 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-n-Myc/MYCN antibody [EPR26107-153] - BSA and Azide free (AB325653)

This data was developed using ab325641, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse large B cell lymphoma tissue labeling n-Myc/MYCN with ab325641 at 1/500 (1.042 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse large B cell lymphoma. The primary antibody was incubated 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IP

Lab

Immunoprecipitation - Anti-n-Myc/MYCN antibody [EPR26107-153] - BSA and Azide free (AB325653)

This data was developed using ab325641, the same antibody clone in a different buffer formulation.

n-Myc/MYCN was immunoprecipitated from 0.35 mg F9 treated with 10 µM MG-132 for 4 hours whole cell lysate with ab325641 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab325641 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : F9 (mouse embryonal carcinoma epithelial cell) treated with 10 µM MG-132 for 4 hours whole cell lysate
Lane 2 : ab325641 IP in F9 treated with 10 µM MG-132 for 4 hours whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab325641 in F9 treated with 10 µM MG-132 for 4 hours whole cell lysate

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

Exposure time : 24 seconds.

All lanes:

Immunoprecipitation - Anti-n-Myc/MYCN antibody [EPR26107-153] (<a href='/en-us/products/primary-antibodies/n-myc-mycn-antibody-epr26107-153-ab325641'>ab325641</a>) at 1/1000 dilution

Lane 1:

F9 (mouse embryonal carcinoma epithelial cell) treated with 10 µM MG-132 for 4 hours whole cell lysate at 10 µg

Lane 2:

<a href='/en-us/products/primary-antibodies/n-myc-mycn-antibody-epr26107-153-ab325641'>ab325641</a> at 1/30 IP in F9 treated with 10 µM MG-132 for 4 hours whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/en-us/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of <a href='/en-us/products/primary-antibodies/n-myc-mycn-antibody-epr26107-153-ab325641'>ab325641</a> in F9 treated with 10 µM MG-132 for 4 hours whole cell lysate

Secondary

All lanes:

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

Observed band size: 60 kDa

false

Exposure time: 24s

• ChIP-seq

Lab

ChIP-sequencing - Anti-n-Myc/MYCN antibody [EPR26107-153] - BSA and Azide free (AB325653)

This data was developed using ab325641, the same antibody clone in a different buffer formulation.

Chromatin was prepared from F9 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab325641 [EPR26107-153]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 70 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-n-Myc/MYCN antibody [EPR26107-153] - BSA and Azide free (AB325653)

This data was developed using ab325641, the same antibody clone in a different buffer formulation.

Chromatin was prepared from F9 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab325641 [EPR26107-153]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 70 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-n-Myc/MYCN antibody [EPR26107-153] - BSA and Azide free (AB325653)

This data was developed using ab325641, the same antibody clone in a different buffer formulation.

Chromatin was prepared from F9 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab325641 [EPR26107-153]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 70 million reads. The Input control is also shown.

• WB

Lab

Western blot - Anti-n-Myc/MYCN antibody [EPR26107-153] - BSA and Azide free (AB325653)

This data was developed using ab325641, the same antibody clone in a different buffer formulation.

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

Low expression : HeLa

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

Exposure time : Lanes 1-2 : 114 seconds, lanes 3-4 : 6 seconds

All lanes:

Western blot - Anti-n-Myc/MYCN antibody [EPR26107-153] (<a href='/en-us/products/primary-antibodies/n-myc-mycn-antibody-epr26107-153-ab325641'>ab325641</a>) at 1/1000 dilution

Lane 1:

IMR-32 (human neuroblastoma neuroblast) whole cell lysate at 40 µg

Lane 2:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 3:

Untreated SK-N-BE(2) (human neuroblastoma neuroblast) whole cell lysate at 40 µg

Lane 4:

SK-N-BE(2) treated with 20 µM MG-132 for 4 hours whole cell lysate at 40 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: 60 kDa,36 kDa

false

• WB

Lab

Western blot - Anti-n-Myc/MYCN antibody [EPR26107-153] - BSA and Azide free (AB325653)

This data was developed using ab325641, the same antibody clone in a different buffer formulation.

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

Low expression : adult hippocampus(PMID : 3049618)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-n-Myc/MYCN antibody [EPR26107-153] (<a href='/en-us/products/primary-antibodies/n-myc-mycn-antibody-epr26107-153-ab325641'>ab325641</a>) at 1/1000 dilution

Lane 1:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 80 µg

Lane 2:

F9 treated with 10 µM MG-132 for 4 hours whole cell lysate at 80 µg

Lane 3:

Mouse E9.5 embryo tissue lysate at 40 µg

Lane 4:

Mouse adult hippocampus tissue lysate at 40 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: 60 kDa,36 kDa

true

Exposure time: 180s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-n-Myc/MYCN antibody [EPR26107-153] - BSA and Azide free (AB325653)

This data was developed using ab325641, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cardiac muscle tissue labeling n-Myc/MYCN with ab325641 at 1/500 (1.042 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on mouse cardiac muscle. The primary antibody was incubated 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-n-Myc/MYCN antibody [EPR26107-153] - BSA and Azide free (AB325653)

This data was developed using ab325641, the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling n-Myc/MYCN with ab325641 at 1/500 (1.042 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Low expression tissue : no staining on mouse cerebrum. The primary antibody was incubated 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 LeicaDS9800 (Bond™ Polymer Refine Detection).

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