Anti-SIK3 antibody [EPR20880-249]

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-SIK3 antibody [EPR20880-249] (AB227044)

Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized Neuro-2a (mouse neuroblastoma neuroblast) cells labelling SIK3 with ab227044 at 1/500 dilution (0.1ug) (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) (ab150077) at 1/2000 dilution was used as the secondary antibody.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-SIK3 antibody [EPR20880-249] (AB227044)

Immunocytochemical analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized Neuro-2a cells labelling SIK3 with ab227044 at 1/50 (15.78 ug/ml) dilution, followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) antibody at 1/1000 dilution (Green). Confocal image showing cytoplasmic staining in Neuro-2a cells. Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) (ab195889) was used to counterstain tubulin at 1/200 (2.5 ug/ml) dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control : Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

• WB

Lab

Western blot - Anti-SIK3 antibody [EPR20880-249] (AB227044)

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

Data was kindly provided by Dr. Qinghua Liu, NIBS.

The 1st Ab working concentration was 5 ug/ml.

All lanes:

Western blot - Anti-SIK3 antibody [EPR20880-249] (ab227044) at 0.789 µg/mL

All lanes:

Mouse cortex tissue lysate at 4 µg

Predicted band size: 140 kDa

Observed band size: 150 kDa,70 kDa

false

• WB

Lab

Western blot - Anti-SIK3 antibody [EPR20880-249] (AB227044)

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

Exposure time : Lane 1 : 3 seconds; Lanes 2-4 : 8 seconds.

All lanes:

Western blot - Anti-SIK3 antibody [EPR20880-249] (ab227044) at 1/1000 dilution

Lane 1:

Rat hippocampus tissue lysate at 20 µg

Lane 2:

Human hippocampus tissue lysate at 20 µg

Lane 3:

Rat liver tissue lysate at 20 µg

Lane 4:

Neuro-2a (mouse neuroblastoma neuroblast) 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 1/20000 dilution

Predicted band size: 140 kDa

Observed band size: 70 kDa

false

• WB

Lab

Western blot - Anti-SIK3 antibody [EPR20880-249] (AB227044)

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

The control and SIK3 knockdown cell lysates were kindly provided by an anonymous collaborator.

Exposure time : Lanes 1-3 : 26 seconds; Lane 4 : 15 seconds.

All lanes:

Western blot - Anti-SIK3 antibody [EPR20880-249] (ab227044) at 1/1000 dilution

Lane 1:

N41 (Embryonic Mouse Hypothalamus Cell Line) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

SIK3 knockdown N41 whole cell lysate at 20 µg

Lane 3:

Mouse hippocampus tissue lysate at 20 µg

Lane 4:

Rat brain tissue 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

Predicted band size: 140 kDa

Observed band size: 70 kDa

false