Anti-NELFe antibody [EPR29850-562]

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NELFe antibody [EPR29850-562] (AB326107)

Immunohistochemical analysis of paraffin-embedded Human colon tissue labeling NELFe with ab326107 at 1/1000 (0.506 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon. 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-NELFe antibody [EPR29850-562] (AB326107)

Immunohistochemical analysis of paraffin-embedded Human colon cancer tissue labeling NELFe with ab326107 at 1/1000 (0.506 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human colon cancer. 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-NELFe antibody [EPR29850-562] (AB326107)

Immunohistochemical analysis of paraffin-embedded Human lung cancer tissue labeling NELFe with ab326107 at 1/1000 (0.506 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on human lung cancer. 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

• ChIP-seq

Lab

ChIP-sequencing - Anti-NELFe antibody [EPR29850-562] (AB326107)

Chromatin was prepared from K-562 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab326107 [EPR29850-562]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-NELFe antibody [EPR29850-562] (AB326107)

Chromatin was prepared from K-562 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab326107 [EPR29850-562]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• ChIP-seq

Lab

ChIP-sequencing - Anti-NELFe antibody [EPR29850-562] (AB326107)

Chromatin was prepared from K-562 cells. Cells were fixed with 1% formaldehyde for 10 minutes. ChIP was performed with 5 x 10^6 cells and 4 µg of ab326107 [EPR29850-562]. ChIP DNA was sequenced on the Illumina NovaSeq 6000 to a depth of 30 million reads. The Input control is also shown.

• IP

Lab

Immunoprecipitation - Anti-NELFe antibody [EPR29850-562] (AB326107)

NELFe was immunoprecipitated from 0.35 mg K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate with ab326107 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab326107 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 2 : ab326107 IP in K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab326107 in K-562 lysate

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

Exposure time : 24 seconds.

To minimize protein degradation, cells/tissues(lane 2, lane 4, lanes 5-8) were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

All lanes:

Immunoprecipitation - Anti-NELFe antibody [EPR29850-562] (ab326107) at 1/1000 dilution

Lane 1:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 10 µg

Lane 2:

ab326107 at 1/30 IP in K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/ko/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab326107 in K-562 lysate

Secondary

All lanes:

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

Observed band size: 43 kDa

false

Exposure time: 24s

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NELFe antibody [EPR29850-562] (AB326107)

Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling NELFe with ab326107 at 1/2000 (0.253 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse colon. 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-NELFe antibody [EPR29850-562] (AB326107)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling NELFe with ab326107 at 1/2000 (0.253 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive 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

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NELFe antibody [EPR29850-562] (AB326107)

Immunohistochemical analysis of paraffin-embedded Mouse breast cancer tissue labeling NELFe with ab326107 at 1/2000 (0.253 µg/ml) dilution, followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Positive staining on mouse breast cancer. 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-NELFe antibody [EPR29850-562] (AB326107)

NELFe was immunoprecipitated from 0.35 mg ES-D3 [D3] (mouse blastocyst-derived embryonic stem cell) whole cell lysate with ab326107 at 1/30 dilution (2µg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab326107 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : ES-D3 [D3] (mouse blastocyst-derived embryonic stem cell) whole cell lysate
Lane 2 : ab326107 IP in ES-D3 [D3] (mouse blastocyst-derived embryonic stem cell) whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab326107 in ES-D3 [D3] lysate

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

Exposure time : 24 seconds.

To minimize protein degradation, cells/tissues(lane 2, lane 4, lanes 5-8) were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

All lanes:

Immunoprecipitation - Anti-NELFe antibody [EPR29850-562] (ab326107) at 1/1000 dilution

Lane 1:

ES-D3 [D3] (mouse blastocyst-derived embryonic stem cell) whole cell lysate at 10 µg

Lane 2:

ab326107 at 1/30 IP in ES-D3 [D3] (mouse blastocyst-derived embryonic stem cell) whole cell lysate

Lane 3:

Rabbit monoclonal IgG (<a href='/ko/products/primary-antibodies/rabbit-igg-monoclonal-epr25a-isotype-control-ab172730'>ab172730</a>) instead of ab326107 in ES-D3 [D3] lysate

Secondary

All lanes:

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

Observed band size: 43 kDa

false

Exposure time: 24s

• WB

Lab

Western blot - Anti-NELFe antibody [EPR29850-562] (AB326107)

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

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) (1 : 200000) (36KDa).

All lanes:

Western blot - Anti-NELFe antibody [EPR29850-562] (ab326107) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

293T transfected with siRNA specifically targeting NELFe whole cell lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 43 kDa,36 kDa

false

Exposure time: 125s

• WB

Lab

Western blot - Anti-NELFe antibody [EPR29850-562] (AB326107)

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

To minimize protein degradation, cells/tissues(lane 2, lane 4, lanes 5-8) 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 (43 kDa) are likely to be degraded target fragments.

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

All lanes:

Western blot - Anti-NELFe antibody [EPR29850-562] (ab326107) at 1/1000 dilution

Lane 1:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 2:

K-562 (human chronic myelogenous leukemia lymphoblast) fresh whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

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

Lane 5:

Mouse brain fresh tissue lysate at 20 µg

Lane 6:

Mouse liver fresh tissue lysate at 20 µg

Lane 7:

Mouse testis fresh tissue lysate at 20 µg

Lane 8:

Mouse spleen fresh tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 43 kDa,36 kDa

false

Exposure time: 180s