Anti-CNTF antibody [EPR24553-104]

• Flow Cyt (Intra)

Supplier Data

Flow Cytometry (Intracellular) - Anti-CNTF antibody [EPR24553-104] (AB270992)

Flow cytometric analysis of 4% paraformaldehyde fixed, 0.1% Tween-20 permeabilised Rat primary neural/glia cells labeling CNTF with ab270992 at 1/500 dilution (0.1μg)/ (Right image) compared with an isotype control Rabbit monoclonal IgG (ab172730)/ (Left image) and an unlabeled control cells without incubation with primary antibody and secondary antibody (Blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

• IHC-Fr

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Immunohistochemistry (Frozen sections) - Anti-CNTF antibody [EPR24553-104] (AB270992)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat sciatic nerve tissue labeling CNTF with ab270992 at 1/100 dilution (4.81 ug/ml) followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on rat sciatic nerve is observed. The nuclear counterstain was DAPI (Blue).

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

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CNTF antibody [EPR24553-104] (AB270992)

Immunohistochemical analysis of paraffin-embedded Mouse stomach tissue labelling CNTF with ab270992 at 1/500 dilution (0.962 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Negative control : No staining in mouse stomach. The section was incubated with ab270992 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CNTF antibody [EPR24553-104] (AB270992)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse sciatic nerve tissue labeling CNTF with ab270992 at 1/100 dilution (4.81 ug/ml) followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Positive staining on mouse sciatic nerve is observed. The nuclear counterstain was DAPI (Blue).

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

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CNTF antibody [EPR24553-104] (AB270992)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Rat stomach tissue labeling CNTF with ab270992 at 1/100 dilution (4.81 ug/ml) followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Negative control : No staining on rat stomach (PMID : 12855189) is observed. The nuclear counterstain was DAPI (Blue).

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

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-Fr

Supplier Data

Immunohistochemistry (Frozen sections) - Anti-CNTF antibody [EPR24553-104] (AB270992)

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse stomach tissue labeling CNTF with ab270992 at 1/100 dilution (4.81 ug/ml) followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (Green). Negative control : No staining on mouse stomach (PMID : 12855189) is observed. The nuclear counterstain was DAPI (Blue).

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

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CNTF antibody [EPR24553-104] (AB270992)

Immunohistochemical analysis of paraffin-embedded Mouse sciatic nerve tissue labelling CNTF with ab270992 at 1/500 dilution (0.962 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining in mouse sciatic nerve. The section was incubated with ab270992 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CNTF antibody [EPR24553-104] (AB270992)

Immunohistochemical analysis of paraffin-embedded Rat sciatic nerve tissue labelling CNTF with ab270992 at 1/500 dilution (0.962 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Cytoplasmic staining in rat sciatic nerve. The section was incubated with ab270992 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins.

• IP

Supplier Data

Immunoprecipitation - Anti-CNTF antibody [EPR24553-104] (AB270992)

CNTF was immunoprecipitated from 0.35 mg Mouse sciatic nerve lysate with ab270992 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270992 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Mouse sciatic nerve lysate 10ug

Lane 2 : ab270992 IP in Mouse sciatic nerve lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab270992 in mouse sciatic nerve lysate

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

Exposure time : 10 seconds.

Bands at 37, 50KD are likely to be multimer forms of CNTF (PMID : 9236223).

All lanes:

Immunoprecipitation - Anti-CNTF antibody [EPR24553-104] (ab270992)

Predicted band size: 23 kDa

Observed band size: 22 kDa,37 kDa,50 kDa

false

• IP

Supplier Data

Immunoprecipitation - Anti-CNTF antibody [EPR24553-104] (AB270992)

CNTF was immunoprecipitated from 0.35 mg Rat sciatic nerve lysate with ab270992 at 1/30 dilution (2ug in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab270992 at 1/1000 dilution. VeriBlot for IP secondary antibody(HRP)(ab131366) was used at 1/5000 dilution.

Lane 1 : Rat sciatic nerve lysate 10ug

Lane 2 : ab270992 IP in Rat sciatic nerve lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab270992 in rat sciatic nerve lysate

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

Exposure time : 10 seconds.

Bands at 40, 75KD are likely to be multimer forms of CNTF (PMID : 9236223).

All lanes:

Immunoprecipitation - Anti-CNTF antibody [EPR24553-104] (ab270992)

Predicted band size: 23 kDa

Observed band size: 22 kDa,37 kDa,50 kDa

false

• WB

Lab

Western blot - Anti-CNTF antibody [EPR24553-104] (AB270992)

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

Bands at 37, 50KD are likely to be multimer forms of CNTF (PMID : 9236223).

Negative control : Brain, stomach (PMID : 12855189).

Exposure time : 59 seconds.

All lanes:

Western blot - Anti-CNTF antibody [EPR24553-104] (ab270992) at 1/1000 dilution

Lane 1:

Mouse sciatic nerve lysate at 20 µg

Lane 2:

Mouse brain lysate at 20 µg

Lane 3:

Mouse stomach 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 1/10000 dilution

Predicted band size: 23 kDa

Observed band size: 22 kDa,37 kDa,50 kDa

false

• WB

Lab

Western blot - Anti-CNTF antibody [EPR24553-104] (AB270992)

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

Negative control : Stomach (PMID : 12855189).

Exposure time : 81 seconds.

All lanes:

Western blot - Anti-CNTF antibody [EPR24553-104] (ab270992) at 1/1000 dilution

Lane 1:

Rat sciatic nerve lysate at 20 µg

Lane 2:

Rat spinal cord lysate at 20 µg

Lane 3:

Rat stomach 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 1/10000 dilution

Predicted band size: 23 kDa

Observed band size: 22 kDa

false