Anti-Chd7 antibody [EPR25355-7] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Chd7 antibody [EPR25355-7] - BSA and Azide free (AB307500)

This data was developed using ab307499, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling CHD7 with ab307499 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-Chd7 antibody [EPR25355-7] - BSA and Azide free (AB307500)

This data was developed using ab307499, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling CHD7 with ab307499 at 1/100 (5.02 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing nuclear staining in SH-SY5Y cell line.Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chd7 antibody [EPR25355-7] - BSA and Azide free (AB307500)

This data was developed using ab307499, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Mouse colon tissue labeling CHD7 with ab307499 at 1/2000 (0.251 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on mouse colon. The section was incubated with ab307499 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) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-Chd7 antibody [EPR25355-7] - BSA and Azide free (AB307500)

This data was developed using ab307499, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized ES-D3 (mouse blastocyst-derived embryonic stem cell) cells labelling CHD7 with ab307499 at 1/100 (5.02 ug/ml) dilution, followed by ab150081 Goat Anti-RABbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2 ug/ml dilution (Green). Confocal image showing nuclear staining in ES-D3 [D3] cell line.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used to counterstain tubulin at 1/200 2.5ug/ml dilution (Red). The Nuclear counterstain was DAPI (Blue). Secondary antibody only control : Secondary antibody is ab150081 Goat Anti-rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/ml dilution.

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-Chd7 antibody [EPR25355-7] - BSA and Azide free (AB307500)

This data was developed using ab307499, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized ES-D3 (mouse blastocyst-derived embryonic stem cell) cells labelling CHD7 with ab307499 at 1/50 dilution (1ug) (Red) compared with a Rabbit monoclonal IgG (ab172730) (Black) isotype control and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (Blue). A Goat Anti-Rabbit IgG (Alexa Fluor^® 488, ab150081) at 1/2000 dilution was used as the secondary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Chd7 antibody [EPR25355-7] - BSA and Azide free (AB307500)

This data was developed using ab307499, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Postnatal day 14 (P1 tissue lABeling CHD7 with ab307499 at 1/2000 (0.251 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on P14 mouse cerebellum. (A) Low powered (magnification, x20) and (B) high powered (magnification, x400) microscopic images. The section was incubated with ab307499 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) was used. 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-Chd7 antibody [EPR25355-7] - BSA and Azide free (AB307500)

This data was developed using ab307499, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Postnatal day 14 (P1 tissue lABeling CHD7 with ab307499 at 1/2000 (0.251 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Nuclear staining on P14 rat cerebellum. The section was incubated with ab307499 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) was used. Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins

• WB

Supplier Data

Western blot - Anti-Chd7 antibody [EPR25355-7] - BSA and Azide free (AB307500)

This data was developed using 307499, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : The bands below 336 kDa may be caused by degradation. Lysates were freshly made and used for Western blotting immediately to minimize protein degradation. Lanes 1-2 : 37 seconds, Lanes 3-5 : 10 seconds. Exposure time :

All lanes:

Western blot - Anti-CHD7 antibody [EPR25355-7] (<a href='/en-us/products/primary-antibodies/chd7-antibody-epr25355-7-ab307499'>ab307499</a>) at 1/1000 dilution

Lane 1:

SH-SY5Y (human neuroblastoma epithelial cell) transfected with scrambled siRNA control whole cell lysate 20 μg

Lane 2:

SH-SY5Y transfected with siRNA specifically targeti CHD7 whole cell lysate 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 1/100000 dilution

Observed band size: 336 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Chd7 antibody [EPR25355-7] - BSA and Azide free (AB307500)

This data was developed using 307499, the same antibody clone in a different buffer formulation.

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

The bands below 336 kDa may be caused by degradation.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

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

All lanes:

Western blot - Anti-CHD7 antibody [EPR25355-7] (<a href='/en-us/products/primary-antibodies/chd7-antibody-epr25355-7-ab307499'>ab307499</a>) at 1/1000 dilution

All lanes:

ES-D3 (mouse embryonic pluripotent stem Cell) 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 1/100000 dilution

Observed band size: 336 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Chd7 antibody [EPR25355-7] - BSA and Azide free (AB307500)

This data was developed using 307499, the same antibody clone in a different buffer formulation.

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

The bands below 336 kDa may be caused by degradation.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

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

All lanes:

Western blot - Anti-CHD7 antibody [EPR25355-7] (<a href='/en-us/products/primary-antibodies/chd7-antibody-epr25355-7-ab307499'>ab307499</a>) at 1/1000 dilution

All lanes:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate 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 1/100000 dilution

Observed band size: 336 kDa

true

Exposure time: 180s