Anti-CNTFR antibody [EPR24672-25]

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CNTFR antibody [EPR24672-25] (AB305245)

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling CNTFR with ab305245 at 1/50 dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green). Confocal image showing mainly membranous staining in SH-SY5Y cell line. Negative control : U-87 MG (PMID : 23061382). 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 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 dilution.

• Flow Cyt

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Flow Cytometry - Anti-CNTFR antibody [EPR24672-25] (AB305245)

Flow cytometric analysis of U-87 MG (human glioblastoma-astrocytoma epithelial cell, Left) / SH-SY5Y (human neuroblastoma epithelial cell, Right) cells labelling CNTFR with ab305245 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). Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Gated on viable cells.Negative control : U-87 MG (PMID : 23061382).

• IP

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Immunoprecipitation - Anti-CNTFR antibody [EPR24672-25] (AB305245)

CNTFR was immunoprecipitated from 0.35 mg SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate 10 μg with ab305245 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab305245. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate 10 μg

Lane 2 : ab305245 at 1/30 IP in SH-SY5Y whole cell lysate 10 μg

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab305245 in SH-SY5Y whole cell lysate

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

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Immunoprecipitation - Anti-CNTFR antibody [EPR24672-25] (ab305245) at 1/30 dilution

All lanes:

SH-SY5Y (human neuroblastoma epithelial cell)

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: 60 kDa

true

Exposure time: 10s

• WB

Supplier Data

Western blot - Anti-CNTFR antibody [EPR24672-25] (AB305245)

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

Low expression : heart, kidney, spleen (PMID : 1648265)

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-CNTFR antibody [EPR24672-25] (ab305245) at 1/1000 dilution

Lane 1:

Human cerebellum tissue lysate at 20 µg

Lane 2:

Human heart tissue lysate at 20 µg

Lane 3:

Human kidney tissue lysate at 20 µg

Lane 4:

Human spleen 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

Observed band size: 60 kDa

true

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-CNTFR antibody [EPR24672-25] (AB305245)

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

Negative control : U-87 MG (PMID : 23061382)

This blot was developed using a high sensitivity ECL substrate. The high-sensitivity ECL substrate used allows for the detection of proteins in the mid-femtogram range.

All lanes:

Western blot - Anti-CNTFR antibody [EPR24672-25] (ab305245) at 1/1000 dilution

Lane 1:

SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate at 20 µg

Lane 2:

U-87 MG (human glioblastoma-astrocytoma 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: 60 kDa

true

Exposure time: 180s

• WB

Lab

Western blot - Anti-CNTFR antibody [EPR24672-25] (AB305245)

Blocking and diluting buffer and concentration : 5% NFDM /TBST. ab181602 was used as a GAPDH loading control at 1/1000000 dilution. Exposure time : Lane 1 : 180 seconds Lane 2 : 20 seconds The sensitivity of ab305245 is lower than that of ab283682, we recommend customers to choose ab283682 when performing WB.

Lane 1:

Western blot - Anti-CNTFR antibody [EPR24672-25] (ab305245) at 1/1000 dilution

Lane 2:

Western blot - Anti-CNTFR antibody [EPR24672-166] (<a href='/en-us/products/primary-antibodies/cntfr-antibody-epr24672-166-ab283682'>ab283682</a>) at 1/1000 dilution

All lanes:

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 1/20000 dilution

Observed band size: 60 kDa

false