重组Anti-SMN/Gemin 1抗体[EPR4429]

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-SMN/Gemin 1 antibody [EPR4429] (AB108531)

IHC image of SMN/Gemin 1 staining in a section of human normal frozen hippocampus* performed on a Leica Biosystems BOND® RX instrument using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab108531, 2.5ugml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-SMN/Gemin 1 antibody [EPR4429] (AB108531)

IHC image of SMN/Gemin 1 staining in a section of human normal frozen cervix* performed on a Leica Biosystems BOND® RX instrument using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab108531, 1ugml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-SMN/Gemin 1 antibody [EPR4429] (AB108531)

IHC image of SMN/Gemin 1 staining in a section of human normal frozen prostate cancer* performed on a Leica Biosystems BOND® RX instrument using the standard protocol. The section was fixed in 10% paraformaldehyde (10 min) prior to staining. The section was incubated with ab108531, 1ugml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset secondary-only control image is taken from an identical assay without primary antibody. *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-SMN/Gemin 1 antibody [EPR4429] (AB108531)

Immunocytochemistry/Immunofluorescence analysis of SH-SY5Y cells labelling Gemin 1 with purified ab108531 at 1/250. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077 an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291 a mouse anti-tubulin (1/500) and ab150120 an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500) were also used.

Control 1 : primary antibody (1/100) and secondary antibody ab150120 an Alexa Fluor^® 594-conjugated goat anti-mouse IgG (1/500).

Control 2 : ab7291 (1/1000) and secondary antibody ab150077 an Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/500).

• WB

Unknown

Western blot - Anti-SMN/Gemin 1 antibody [EPR4429] (AB108531)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-SMN/Gemin 1 antibody [EPR4429] (ab108531) at 1/5000 dilution

All lanes:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 32 kDa

Observed band size: 35 kDa

false

• WB

Lab

Western blot - Anti-SMN/Gemin 1 antibody [EPR4429] (AB108531)

Western blot : Rabbit Monoclonal [EPR4429] to SMN/Gemin 1 ab108531 staining at 1/1000 dilution, shown in green; Mouse anti alpha Tubulin (ab7291) loading control staining at 1/20,000 dilution, shown in magenta.

A band was observed at 32 kDa in Wild-type A549 cell lysates with no signal observed at this size in SMN1 knockout A549 cell line.

To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3pc Milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged.

Secondary antibodies used were Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-SMN/Gemin 1 antibody [EPR4429] (ab108531) at 1/1000 dilution

Lane 1:

Wild-type A549 at 20 µg

Lane 2:

SMN1 knockout A549 at 20 µg

Lane 2:

Western blot - Human SMN1 knockout A549 cell line (<a href='/products/cell-lines/human-smn1-knockout-a549-cell-line-ab301247'>ab301247</a>) at 20 µg

Lane 3:

HeLa at 20 µg

Lane 4:

PC-3 at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 32 kDa

Observed band size: 32 kDa

false

• WB

Unknown

Western blot - Anti-SMN/Gemin 1 antibody [EPR4429] (AB108531)

All lanes:

Western blot - Anti-SMN/Gemin 1 antibody [EPR4429] (ab108531) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

HepG2 cell lysate at 10 µg

Lane 3:

K562 cell lysate at 10 µg

Lane 4:

293T cell lysate at 10 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

Predicted band size: 32 kDa

false

• WB

Unknown

Western blot - Anti-SMN/Gemin 1 antibody [EPR4429] (AB108531)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-SMN/Gemin 1 antibody [EPR4429] (ab108531) at 1/1000 dilution

All lanes:

HeLa cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 105 kDa,11 kDa,32 kDa

Observed band size: 105 kDa,14 kDa,35 kDa,50 kDa

false

• WB

Unknown

Western blot - Anti-SMN/Gemin 1 antibody [EPR4429] (AB108531)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-SMN/Gemin 1 antibody [EPR4429] (ab108531) at 1/1000 dilution

Lane 1:

K562 cell lysate at 20 µg

Lane 2:

HEK-293 cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 32 kDa

Observed band size: 35 kDa

false