Kidney sections were examined by indirect immunofluorescence on 3-4-micron thick paraffin sections. The sections were deparaffinized with xylene and rehydrated in graded ethanol and antigen retrieval was performed by boiling in 10 mM sodium citrate followed by washes in PBS-Tween (0.5% Tween-20). Sections were blocked with SuperBlock (Scytek Labs Inc., Logan, UT) for 6 min at 37°C, followed by incubation with anti-synaptopodin primary antibody (Santa Cruz Biotechnology) in 5% super-block at 4°C overnight at 1:100 dilution. Sections were washed with 2.5% SuperBlock in PBS-Tween (0.5% Tween-20), three times for 10 min each and incubated with fluorescent secondary antibody (Alexa Flour, 2 ug/ml; Invitrogen, Carlsbad, CA) in 5% SuperBlock in PBS for 1 hour at room temperature. Sections were washed three times for 10 min each with 2.5% SuperBlock in PBS-Tween (0.5% Tween-20) and mounted with Prolong Gold Antifade Reagent (Invitrogen). Kidney sections from the groups were viewed and imaged with equal exposures with BZ-X700 All-in-one fluorescence microscope (Keyence Inc., Itasca, IL).
Anti synaptopodin primary antibody
Manufactured by Santa Cruz Biotechnology
The Anti-synaptopodin primary antibody is a laboratory reagent used for the detection and analysis of the synaptopodin protein in various biological samples. Synaptopodin is a crucial structural component of the podocyte foot processes in the kidney glomerulus. This antibody can be utilized in techniques such as Western blotting, immunohistochemistry, and immunocytochemistry to identify and study the expression and localization of the synaptopodin protein.
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2 protocols using anti synaptopodin primary antibody
1
Immunofluorescence Analysis of Kidney Synaptopodin
2
Indirect Immunofluorescence of Kidney
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Kidneys were examined by indirect immunofluorescence on 4-micron thick paraffin sections. The sections were deparaffinized with xylene and rehydrated in graded ethanol. Antigen retrieval was performed by boiling the slides in 10 mM sodium citrate followed by PBS-Tween (0.5% Tween-20) wash. Blocking was done with Super Block (Scytek Labs Inc., Logan, UT) for 6 min at 37 °C, followed by incubation with anti-synaptopodin primary antibody (Santa Cruz Biotechnology) in 5% super-block at 4 °C overnight at 1:100 dilution. Sections were washed three times for 10 min each with 2.5% Super Block in PBS-Tween (0.5% Tween-20) and incubated with fluorescent secondary antibody (Alexa Flour, 2 ug/ml; Invitrogen, Carlsbad, CA) in 5% Super Block in PBS for 1 hour at room temperature. Sections were then washed three times for 10 min each with 2.5% Super Block in PBS-Tween (0.5% Tween-20), mounted with Prolong Gold Antifade Reagent (Invitrogen). Kidney sections from the groups were viewed and imaged with exposures within 250–600 ms with an Axio Scope fluorescent microscope (Carl Zeiss Microscopy GmbH, Jena, Germany).
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