Anti-SMN/Gemin 1 antibody [EPR25897-27] - BSA and Azide free

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-SMN/Gemin 1 antibody [EPR25897-27] - BSA and Azide free (AB314896)

This data was developed using ab314895, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling SMN/Gemin 1 with ab314895 at 1/500 dilution (0.1 ug)/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 1/5000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SMN/Gemin 1 antibody [EPR25897-27] - BSA and Azide free (AB314896)

This data was developed using ab314895, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized HeLa (human cervical adenocarcinoma epithelial cell) cells labelling SMN/Gemin 1 with ab314895 at 1/50 (10.34 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 and cytoplasmic staining in HeLa cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). 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.

• IP

Supplier Data

Immunoprecipitation - Anti-SMN/Gemin 1 antibody [EPR25897-27] - BSA and Azide free (AB314896)

This data was developed using ab314895, the same antibody clone in a different buffer formulation.

SMN/Gemin 1 was immunoprecipitated from 0.35 mg HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate with ab314895 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314895 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate

Lane 2 : ab314895 IP in HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate

Lane 3 : Rabbit monoclonal lgG (ab172730) instead of ab314895 in HeLa whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-SMN/Gemin 1 antibody [EPR25897-27] (<a href='/en-us/products/primary-antibodies/smn-gemin-1-antibody-epr25897-27-ab314895'>ab314895</a>) at 1/30 dilution

All lanes:

HeLa (human cervix adenocarcinoma epithelial cell) 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

false

Exposure time: 3s

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SMN/Gemin 1 antibody [EPR25897-27] - BSA and Azide free (AB314896)

This data was developed using ab314895, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling SMN/Gemin 1 with ab314895 at 1/50 (10.34 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 and cytoplasmic staining in Neuro-2a cell line. Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8). 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-SMN/Gemin 1 antibody [EPR25897-27] - BSA and Azide free (AB314896)

This data was developed using ab314895, the same antibody clone in a different buffer formulation.

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling SMN/Gemin 1 with ab314895 at 1/500 dilution (0.1 ug)/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 1/5000 dilution was used as the secondary antibody.

• IP

Supplier Data

Immunoprecipitation - Anti-SMN/Gemin 1 antibody [EPR25897-27] - BSA and Azide free (AB314896)

This data was developed using ab314895, the same antibody clone in a different buffer formulation.

SMN/Gemin 1 was immunoprecipitated from 0.35 mg Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate with ab314895 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab314895 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate

Lane 2 : ab314895 IP in Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab314895 in Neuro-2a whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-SMN/Gemin 1 antibody [EPR25897-27] (<a href='/en-us/products/primary-antibodies/smn-gemin-1-antibody-epr25897-27-ab314895'>ab314895</a>) at 1/30 dilution

All lanes:

Neuro-2a (mouse neuroblastoma neuroblast) 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

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-SMN/Gemin 1 antibody [EPR25897-27] - BSA and Azide free (AB314896)

This data was developed using ab314895, the same antibody clone in a different buffer formulation.

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

Lane 1-2 and lane 4-5 were freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-SMN/Gemin 1 antibody [EPR25897-27] (<a href='/en-us/products/primary-antibodies/smn-gemin-1-antibody-epr25897-27-ab314895'>ab314895</a>) at 1/1000 dilution

Lane 1:

HeLa (human epithelial cell line from cervical adenocarcinoma) whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

SH-SY5Y (human neuroblastoma epithelial 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: 35 kDa

false

Exposure time: 180s