Anti-UNC5B antibody [EPR18439] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-UNC5B antibody [EPR18439] - BSA and Azide free (AB251002)

This data was developed using ab189914, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized C6 (Rat glial tumor cell line) cells labeling UNC5B with ab189914 at 1/500 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing cytoplasm staining on C6 cell line.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody - Loading Control (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG(AlexaFluor®594) preadsorbed (ab150120) at 1/1000 dilution (red).

The negative controls are as follows : -

-ve control 1 : ab189914 at 1/500 dilution followed by ab150120 at 1/1000 dilution.

-ve control 2 : ab7291 at 1/1000 dilution followed by ab150077 at 1/1000 dilution.

• WB

Supplier Data

Western blot - Anti-UNC5B antibody [EPR18439] - BSA and Azide free (AB251002)

This data was developed using ab189914, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

UNC5B was cleaved by caspase-3 at Asp412 to produce a cleavage fragment around ~60kDa Llambi F. et al. EMBO J. 2001 Jun 1; 20(11) : 2715-22.

All lanes:

Western blot - Anti-UNC5B antibody [EPR18439] (<a href='/en-us/products/primary-antibodies/unc5b-antibody-epr18439-ab189914'>ab189914</a>) at 1/10000 dilution

All lanes:

C6 (Rat glial tumor cell line) whole cell lysate at 10 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 1/50000 dilution

Predicted band size: 103 kDa

Observed band size: 130 kDa,60 kDa

false

Exposure time: 2min

• IP

Supplier Data

Immunoprecipitation - Anti-UNC5B antibody [EPR18439] - BSA and Azide free (AB251002)

This data was developed using ab189914, the same antibody clone in a different buffer formulation.

UNC5B was immunoprecipitated from 1mg of Rat cerebral cortex whole cell extract with ab189914 at 1/50 dilution.

Western blot was performed from the immunoprecipitate using ab189914 at 1/1000 dilution.

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/500 dilution.

Lane 1 : Rat cerebral cortex whole cell extract,10μg (Input).
Lane 2 : ab189914 IP in Rat cerebral cortex whole cell extract.
Lane 3 : Rabbit IgG,monoclonal - Isotype Control (ab172730) instead of ab189914 in Rat cerebral cortex whole cell extract.

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

Exposure time : 30 seconds.

All lanes:

Immunoprecipitation - Anti-UNC5B antibody [EPR18439] (<a href='/en-us/products/primary-antibodies/unc5b-antibody-epr18439-ab189914'>ab189914</a>)

Predicted band size: 103 kDa

false

• WB

Supplier Data

Western blot - Anti-UNC5B antibody [EPR18439] - BSA and Azide free (AB251002)

This data was developed using ab189914, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

UNC5B was cleaved by caspase-3 at Asp412 to produce a cleavage fragment around ~60kDa Llambi F. et al. EMBO J. 2001 Jun 1; 20(11) : 2715-22.

All lanes:

Western blot - Anti-UNC5B antibody [EPR18439] (<a href='/en-us/products/primary-antibodies/unc5b-antibody-epr18439-ab189914'>ab189914</a>) at 1/2000 dilution

Lane 1:

Rat cerebral cortex tissue lysate at 10 µg

Lane 2:

Rat hippocampus tissue lysate at 10 µg

Lane 3:

Rat hypothalamus tissue lysate at 10 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 1/50000 dilution

Predicted band size: 103 kDa

Observed band size: 130 kDa,60 kDa

false

Exposure time: 10s