Anti-RDH5 antibody [EPR15191] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-RDH5 antibody [EPR15191] - BSA and Azide free (AB251299)

This data was developed using ab200197, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized Y79 (Human retinoblastoma cell line) cells labeling RDH5 with ab200197 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/500 dilution (green).

Cytoplasm staining on Y79 cell line is observed.

The nuclear counterstain is DAPI (blue).

Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution and ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution (red).

The negative controls are as follows : -
-ve control 1 : ab200197 at 1/100 dilution followed by ab150120 (AlexaFluor®594 Goat anti-Mouse secondary) at 1/500 dilution.
-ve control 2 : ab7291 (anti-Tubulin mouse mAb) at 1/1000 dilution followed by ab150077 (Alexa Fluor®488 Goat Anti-Rabbit IgG H&L) at 1/500 dilution.

• WB

Supplier Data

Western blot - Anti-RDH5 antibody [EPR15191] - BSA and Azide free (AB251299)

This data was developed using ab200197, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-RDH5 antibody [EPR15191] (<a href='/en-us/products/primary-antibodies/rdh5-antibody-epr15191-ab200197'>ab200197</a>) at 1/200 dilution

All lanes:

Y79 (Human retinoblastoma cell line) cell lysate at 30 µg

Secondary

All lanes:

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

Predicted band size: 35 kDa

Observed band size: 35 kDa

false

Exposure time: 3min

• IP

Supplier Data

Immunoprecipitation - Anti-RDH5 antibody [EPR15191] - BSA and Azide free (AB251299)

This data was developed using ab200197, the same antibody clone in a different buffer formulation.

RDH5 was immunoprecipitated from 1mg of Rat eyeball whole cell lysate with ab200197 at 1/30 dilution.

Lane 1 : Rat eyeball whole cell lysate 10ug (Input).

Lane 2 : ab200197 IP in Rat eyeball whole cell lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab200197 in Rat eyeball whole cell lysate.

Western blot was performed from the immunoprecipitate using ab200197 at 1/1000 dilution.

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG, was used as secondary antibody at 1/1500.

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

Exposure time : 10 seconds.

All lanes:

Immunoprecipitation - Anti-RDH5 antibody [EPR15191] (<a href='/en-us/products/primary-antibodies/rdh5-antibody-epr15191-ab200197'>ab200197</a>)

Predicted band size: 35 kDa

false

• WB

Supplier Data

Western blot - Anti-RDH5 antibody [EPR15191] - BSA and Azide free (AB251299)

This data was developed using ab200197, the same antibody clone in a different buffer formulation.

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-RDH5 antibody [EPR15191] (<a href='/en-us/products/primary-antibodies/rdh5-antibody-epr15191-ab200197'>ab200197</a>) at 1/5000 dilution

Lane 1:

Mouse eyeball tissue lysate at 10 µg

Lane 2:

Rat eyeball tissue lysate at 10 µg

Lane 3:

Mouse spleen tissue lysate at 10 µg

Lane 4:

Rat spleen tissue lysate at 10 µg

Lane 5:

PC-12 (Rat adrenal gland pheochromocytoma) cell lysate at 10 µg

Lane 6:

NIH/3T3 (Mouse embyro fibroblast cells) cell lysate at 10 µg

Secondary

All lanes:

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

Predicted band size: 35 kDa

Observed band size: 35 kDa

false

Exposure time: 15s