Immunohistochemistry Customer Review Anti-Smo antibody (STJ95710)
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G protein-coupled receptor that probably associates with the patched protein (PTCH) to transduce the hedgehog's proteins signal. Binding of sonic hedgehog (SHH) to its receptor patched is thought to prevent normal inhibition by patched of smoothened (SMO). Required for the accumulation of KIF7 and GLI3 in the cilia. Anti-Smo antibody (STJ95710): http://www.stjohnslabs.com/smo-antibody-p-94371?filter_name=STJ95710 Join our Antibody Validation Project - http://www.stjohnslabs.com/services/antibody-validation
![Rating:
Specificity:
ANTIBODY CUSTOMER REVIEW
Findings: “Antibody works well in DAB
immunohistochemistry at 1:500 dilution.” .-Tristan Varela, KCL
Knowledge Dock Business Centre, University Way, London E16 2RD
T: (+44) 0208 223 3081 F: (+44) 0207 681 2580 E: info@stjohnslabs.com w: www.stjohnslabs.com
Protocol
Rehydration: Slides placed in xylene for dewaxing
[2 x 10minutes]
Rehydrated with EtOH: 2 x 100%
Etoh, 70% EtOH, 50% EtOH [5
minutes each]
Washing Wash slides in 0.5 Tris-buffered Saline
solution pH 7.6 [2 x 10 minutes]
Washing
Wash briefly in ddH2O
Wash in 3% Hydrogen Peroxide [15
minutes]
Wash briefly in ddH2O
Secondary
Antibody
Probing &
Wash
Add HRP-conjugated secondary
antibody diluted in block solution and
incubate at room temperature [1 hour]
Wash slides in 0.5 Tris-buffered Saline
solution pH 7.6 [2 x 10 minutes]
Heating Heat slides in a pressure cooker in
antigen retrieval solution (10mM
Tris Base, 1mM EDTA Solution,
0.05% Tween 20, pH 9.0) [25
minutes] Encircle sections with a
PAP pen, and add block solution
(1% Bovine serum albumin, 0.05M
Tris-buffered Saline, 0.05% Sodium
Azide, pH 7.6) [30 mins]
DAB
Staining
And Wash
Incubate in DAB solution (D4418
SIGMA) [15 minutes]. Wash briefly in
ddH2O. Dehydrate with EtOH: 50%
EtOH, 70% EtOH, 2 x 100% Etoh [5
minutes each]. Xylene [2 x 10
minutes]. Mount slides in DPX and
leave to dry overnight
Primary Antibody
Probing
Remove block and add primary
antibody diluted in block solution to
sections.
Incubate overnight at 4oC
Visualisation Confocal Microscopy
Model Number STJ95710
Antibody Name Anti-Smo antibody
Application Immunohistochemistry
Species Chicken
Tissue E16 chicken embryo
cerebellum wax section
Dilution 1:500
Secondary Antibody HRP goat anti-rabbit
Figure 1. The expression of Smoothened has not
been well established in the cerebellum but this
antibody appears to be staining a highly specific
cell population in the cerebellum.
Morphologically, it is likely that these are
Bergmann Glia cells, but further analysis would
have to be performed in order to confirm this](https://image.slidesharecdn.com/stj95710anti-smoantibody-170628144813/85/immunohistochemistry-customer-review-antismo-antibody-stj95710-1-320.jpg)
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Immunohistochemistry Customer Review Anti-Smo antibody (STJ95710)
- 1. Rating: Specificity: ANTIBODY CUSTOMER REVIEW Findings: “Antibody works well in DAB immunohistochemistry at 1:500 dilution.” .-Tristan Varela, KCL Knowledge Dock Business Centre, University Way, London E16 2RD T: (+44) 0208 223 3081 F: (+44) 0207 681 2580 E: info@stjohnslabs.com w: www.stjohnslabs.com Protocol Rehydration: Slides placed in xylene for dewaxing [2 x 10minutes] Rehydrated with EtOH: 2 x 100% Etoh, 70% EtOH, 50% EtOH [5 minutes each] Washing Wash slides in 0.5 Tris-buffered Saline solution pH 7.6 [2 x 10 minutes] Washing Wash briefly in ddH2O Wash in 3% Hydrogen Peroxide [15 minutes] Wash briefly in ddH2O Secondary Antibody Probing & Wash Add HRP-conjugated secondary antibody diluted in block solution and incubate at room temperature [1 hour] Wash slides in 0.5 Tris-buffered Saline solution pH 7.6 [2 x 10 minutes] Heating Heat slides in a pressure cooker in antigen retrieval solution (10mM Tris Base, 1mM EDTA Solution, 0.05% Tween 20, pH 9.0) [25 minutes] Encircle sections with a PAP pen, and add block solution (1% Bovine serum albumin, 0.05M Tris-buffered Saline, 0.05% Sodium Azide, pH 7.6) [30 mins] DAB Staining And Wash Incubate in DAB solution (D4418 SIGMA) [15 minutes]. Wash briefly in ddH2O. Dehydrate with EtOH: 50% EtOH, 70% EtOH, 2 x 100% Etoh [5 minutes each]. Xylene [2 x 10 minutes]. Mount slides in DPX and leave to dry overnight Primary Antibody Probing Remove block and add primary antibody diluted in block solution to sections. Incubate overnight at 4oC Visualisation Confocal Microscopy Model Number STJ95710 Antibody Name Anti-Smo antibody Application Immunohistochemistry Species Chicken Tissue E16 chicken embryo cerebellum wax section Dilution 1:500 Secondary Antibody HRP goat anti-rabbit Figure 1. The expression of Smoothened has not been well established in the cerebellum but this antibody appears to be staining a highly specific cell population in the cerebellum. Morphologically, it is likely that these are Bergmann Glia cells, but further analysis would have to be performed in order to confirm this