Anti-HuD antibody [EPR26472-54] (BSA and Azide free)

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HuD antibody [EPR26472-54] (BSA and Azide free) (AB302515)

This data was developed using ab302514, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human pancreas tissue labeling HuD with ab302514 at 1/500 (1.054 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on islet in human pancreas at a low level (PMID : 22387028). The section was incubated with ab302514 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) was used. 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-HuD antibody [EPR26472-54] (BSA and Azide free) (AB302515)

This data was developed using ab302514, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human cerebellum tissue labeling HuD with ab302514 at 1/500 (1.054 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human cerebellum. The section was incubated with ab302514 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) was used. 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-HuD antibody [EPR26472-54] (BSA and Azide free) (AB302515)

This data was developed using ab302514, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling HuD with ab302514 at 1/500 (1.054 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human cerebrum (PMID : 20584986). The section was incubated with ab302514 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) was used. 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-HuD antibody [EPR26472-54] (BSA and Azide free) (AB302515)

This data was developed using ab302514, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Panel A. HEK-293T ce tissue labeling HuD with ab302514 at 1/5000 (0.105 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on HEK-293T cells transfected with a human HuD expression vector (Panel A); No staining on HEK-293T cells transfected with a human HuC expression vector (Panel B); No staining on HEK-293T cells transfected with a human HuB expression vector (Panel C); No staining on HEK-293T cells transfected with an empty vector (Panel D).The section was incubated with ab302514 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) was used. 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-HuD antibody [EPR26472-54] (BSA and Azide free) (AB302515)

This data was developed using ab302514, the same antibody clone in a different buffer formulation. Immunohistochemical analysis of paraffin-embedded Human glioma tissue labeling HuD with ab302514 at 1/500 (1.054 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection) was used. Positive staining on human glioma. The section was incubated with ab302514 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) was used. Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-HuD antibody [EPR26472-54] (BSA and Azide free) (AB302515)

This data was developed using ab302514, the same antibody clone in a different buffer formulation. Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized SH-SY5Y (human neuroblastoma epithelial cell) cells labelling HuD with ab302514 at 1/500 (1.054 ug/ml) dilution, followed by ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 2ug/mL dilution (Green). Confocal image showing cytoplasmic and nuclear staining in SH-SY5Y cell line.Negative control : K-562 (PMID : 25523825) 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 2ug/mL dilution.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-HuD antibody [EPR26472-54] (BSA and Azide free) (AB302515)

This data was developed using ab302514, the same antibody clone in a different buffer formulation. Flow cytometric analysis of 4% paraformaldehyde fixed 90% methanol permeabilized K562 (human chronic myelogenous leukemia lymphoblast, Left) / SH-SY5Y (human neuroblastoma epithelial cell, Right) cells labelling HuD with ab302514 at 1/500 dilution (0.1ug) (Red) (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). A Goat Anti-Rabbit IgG (Alexa Fluor® 488, ab150081) at 1/2000 dilution was used as the secondary antibody. Negative control : K-562 (PMID : 25523825).

• IP

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Immunoprecipitation - Anti-HuD antibody [EPR26472-54] (BSA and Azide free) (AB302515)

This data was developed using ab302514, the same antibody clone in a different buffer formulation.

HuD was immunoprecipitated from 0.35 mg SH-SY5Y (human neuroblastoma epithelial cell), whole cell lysate 10 μg with ab302514 at 1/30 dilution (2μg in 0.35mg lysates). Western blot was performed on the immunoprecipitate using ab302514 at 1/1000 dilution. 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 (Inset)

Lane 2 : ab302514 IP in SH-SY5Y whole cell lysate

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab302514 in SH-SY5Y whole cell lysate

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

All lanes:

Immunoprecipitation - Anti-HuD antibody [EPR26472-54] (<a href='/en-us/products/primary-antibodies/hud-antibody-epr26472-54-ab302514'>ab302514</a>)

All lanes:

SH-SY5Y (human neuroblastoma epithelial cell), 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

Predicted band size: 42 kDa

false

Exposure time: 15s

• WB

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Western blot - Anti-HuD antibody [EPR26472-54] (BSA and Azide free) (AB302515)

This data was developed using ab302514, the same antibody clone in a different buffer formulation. Blocking and diluting buffer and concentration : 5% NFDM/TBST Negative control : K-562 (PMID : 25523825).

All lanes:

Western blot - Anti-HuD antibody [EPR26472-54] (<a href='/en-us/products/primary-antibodies/hud-antibody-epr26472-54-ab302514'>ab302514</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

K-562 (human chronic myelogenous leukemia lymphoblast), 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: 40 kDa

false

Exposure time: 3min