Anti-NSUN6 antibody [EPR27179-64] - BSA and Azide free

• IP

Supplier Data

Immunoprecipitation - Anti-NSUN6 antibody [EPR27179-64] - BSA and Azide free (AB307431)

This data was developed using ab307430, the same antibody clone in a different buffer formulation.

NSUN6 was immunoprecipitated from 0.35 mg Hela (human cervix adenocarcinoma epithelial cell) whole cell lysate 20 ug with ab307430 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab307430 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 20 ug
Lane 2 : ab307430 IP in Hela whole cell lysate
Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab307430 in Hela whole cell lysate

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

Exposure time : 84 seconds

All lanes:

Immunoprecipitation - Anti-NSUN6 antibody [EPR27179-64] (<a href='/en-us/products/primary-antibodies/nsun6-antibody-epr27179-64-ab307430'>ab307430</a>) at 1/1000 dilution

Lane 1:

Hela (human cervix adenocarcinoma epithelial cell) whole cell lysate 20 μg

Lane 2:

Hela 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

Observed band size: 50 kDa

false

Exposure time: 84s

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-NSUN6 antibody [EPR27179-64] - BSA and Azide free (AB307431)

This data was developed using ab307430, 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 NSUN6 with ab307430 at 1/100 (4.81 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 cytoplasmic staining in Neuro-2a cell line.Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8). is observed. 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.

• WB

Supplier Data

Western blot - Anti-NSUN6 antibody [EPR27179-64] - BSA and Azide free (AB307431)

This data was developed using 307430, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : Low expression : human skeletal muscle (PMID : 33330931). The identity of the higher MW band at approximately 75 kDa is unknown. Exposure time :

All lanes:

Western blot - Anti-NSUN6 antibody [EPR27179-64] (<a href='/en-us/products/primary-antibodies/nsun6-antibody-epr27179-64-ab307430'>ab307430</a>) at 1/1000 dilution

Lane 1:

Human testis tissue lysate 20 μg

Lane 2:

Human skeletal muscle tissue lysate 20 μg

Lane 3:

Mouse brain tissue lysate 20 μg

Lane 4:

Mouse embryo tissue lysate 20 μg

Lane 5:

Rat testis tissue lysate 20 μg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 50 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-NSUN6 antibody [EPR27179-64] - BSA and Azide free (AB307431)

This data was developed using 307430, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 125 seconds Exposure time :

All lanes:

Western blot - Anti-NSUN6 antibody [EPR27179-64] (<a href='/en-us/products/primary-antibodies/nsun6-antibody-epr27179-64-ab307430'>ab307430</a>) at 1/1000 dilution

Lane 1:

Hela (human cervix adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate 20 μg

Lane 2:

Hela transfected with siRNA specifically targeti NSUN6 whole cell lysate 20 μg

Lane 3:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate 20 μg

Lane 4:

HepG2 (human hepatocellar carcinoma epithelial cell lysate 20 μg

Lane 5:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate 20 μg

Lane 6:

Hela (human cervix adenocarcinoma epithelial cell) whole cell lysate 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: 50 kDa

false

Exposure time: 125s