Anti-Sonic Hedgehog 抗体 [EP1190Y]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sonic Hedgehog antibody [EP1190Y] (AB53281)

Immunohistochemical analysis of Formalin fixed paraffin-embedded human kidney cancerous tissue labeling Sonic Hedgehog with unpurified ab53281 at 1/100 dilution.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Sonic Hedgehog antibody [EP1190Y] (AB53281)

Overlay histogram showing HepG2 cells stained with unpurified ab53281 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab53281, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight^® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HepG2 cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

• IHC-P

PubMed

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sonic Hedgehog antibody [EP1190Y] (AB53281)

Immunohistochemical analysis of Formalin fixed paraffin embedded human ovarian cancer tissue, staining Sonic Hedgehog with unpurified ab53281.

Antigen retrieval was carried out in citrate buffer using a pressure cooker for 40 minutes. Sections were blocked with blocking agent before incubating with primary antibody (1/2000) for 90 minutes at room temperature. Staining was detected using DAB.

Image from McCann CK et al. PLoS One. 2011;6(11):e28077. Epub 2011 Nov 29. Fig 1.; doi:10.1371/journal.pone.0028077; November 29 2011 PLoS ONE 6(11): e28077.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Sonic Hedgehog antibody [EP1190Y] (AB53281)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling Sonic Hedgehog with unpurified ab53281 at 1/20 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluorr^®488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Sonic Hedgehog antibody [EP1190Y] (AB53281)

Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) labelling Sonic Hedgehog with purified ab53281 at 1/500. Cells were fixed with 4% Paraformaldehyde and permeabilised by 0.1% tritonX-100. An Alexa Fluor^® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody (ab150077). Nuclei counterstained with DAPI (blue).

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Sonic Hedgehog antibody [EP1190Y] (AB53281)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Sonic Hedgehog with purified ab53281 at 1/250 dilution (0.8μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889, Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1/200 (2.5 μg/ml). ab150077 Goat anti rabbit IgG(Alexa Fluor^® 488) was used as the secondary antibody at 1/1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Sonic Hedgehog antibody [EP1190Y] (AB53281)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human pancreatic carcinoma tissue sections labeling Sonic Hedgehog with purified ab53281 at 1/2000 dilution (0.097 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Tissue was counterstained with Hematoxylin. ab97051 Goat Anti-Rabbit IgG H&L (HRP) secondary antibody was used at 1/500 dilution. PBS instead of the primary antibody was used as the negative control.

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Sonic Hedgehog antibody [EP1190Y] (AB53281)

Immunocytochemistry/Immunofluorescence analysis of HepG2 cells labelling Sonic Hedgehog with unpurified ab53281.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Sonic Hedgehog antibody [EP1190Y] (AB53281)

Intracellular Flow Cytometry analysis of HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Sonic Hedgehog with purified ab53281 at 1/20 dilution (10 ug/ml) (red). Cells were fixed with 4% Paraformaldehyde. A Goat anti rabbit IgG (Alexa Fluorr^® 488) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - cells without incubation with primary antibody and secondary antibody (Blue).

• WB

Unknown

Western blot - Anti-Sonic Hedgehog antibody [EP1190Y] (AB53281)

All lanes:

Western blot - Anti-Sonic Hedgehog antibody [EP1190Y] (ab53281) at 1/2000 dilution

All lanes:

Fetal liver membrane at 10 µg

Secondary

All lanes:

Goat anti-rabbit HRP at 1/2000 dilution

Predicted band size: 49 kDa

Observed band size: 27 kDa,49 kDa,51 kDa

false

• WB

Lab

Western blot - Anti-Sonic Hedgehog antibody [EP1190Y] (AB53281)

Blocking and diluting buffer : 5% NFDM /TBST .

27KDa : C-terminal product.

All lanes:

Western blot - Anti-Sonic Hedgehog antibody [EP1190Y] (ab53281) at 1/1000 dilution

All lanes:

Human fetal kidney lysate at 15 µg

Secondary

All lanes:

Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

Predicted band size: 49 kDa

Observed band size: 27 kDa

false

• WB

Lab

Western blot - Anti-Sonic Hedgehog antibody [EP1190Y] (AB53281)

Blocking and diluting buffer : 5% NFDM/TBST.

50kDa : Full-length SHH; 27KDa : C-terminal product

All lanes:

Western blot - Anti-Sonic Hedgehog antibody [EP1190Y] (ab53281) at 1/2000 dilution

All lanes:

Human fetal liver lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 49 kDa

Observed band size: 27 kDa,50 kDa

false

• WB

CiteAb

Western blot - Anti-Sonic Hedgehog antibody [EP1190Y] (AB53281)

Sonic Hedgehog western blot using anti-Sonic Hedgehog antibody [EP1190Y] ab53281. Publication image and figure legend from Wang, Z., Jiang, J., et al., 2019, J Cell Mol Med, PubMed 30761741.

ab53281 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab53281 please see the product overview.

The expression of EIF5A controls the sHH signalling pathway in PC cells. The Western blot showed that EIF5A knockdown can decrease the protein expression of sHH signalling factors in Panc-1 cells (A) and BxPc-3 cells (B). Graphs showed the EIF5A knockdown obviously reduced the expressions of sHH and Gli-1 in Panc-1 cells (C) and BxPc-3 cells (D). To decide whether activation of sHH signalling pathway depends on sHH factor, we used recombinant sHH or neutralizing antibody to treat the PC cells with Si-EIF5A. The Western blot results showed the low expression level of Gli-1 for Panc-1 cells (E) and for BxPc-3 cells (F). *p < 0.05 as determined by the Student's t test

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