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Anti-GFAP [N206A/8R] in Immunocytochemistry (Rat)

Addgene #193445

Data Submitted By
Zuzanna Rauk, Amanda Della Giustina, Kristina Sakers and Guiliana Pellegrino
Lab Name
CAJAL training programme
Submission Date
February 06, 2024
Publication Date
February 06, 2024 (modified October 15, 2025)
DOI
https://doi.org/10.57733/addgene.yzfsry
Abstract
Experimental results for Anti-GFAP [N206A/8R] (Addgene Antibody 193445) in an immunocytochemistry assay with a rat sample.
Citation
Anti-GFAP [N206A/8R] in Immunocytochemistry (Rat). Rauk Z, Della Giustina A, Sakers K, Pellegrino G 2024. Addgene Report, https://doi.org/10.57733/addgene.yzfsry
Negative control for Addgene antibody # 193445: Rat astrocytes treated with blocking solution, PBST + NGS instead of primary antibody, and Alexa 546 conjugated secondary anti-mouse antibody.
Caption: Negative control for Addgene antibody # 193445: Rat astrocytes treated with blocking solution, PBST + NGS instead of primary antibody, and Alexa 546 conjugated secondary anti-mouse antibody.
Image attribution: Zuzanna Rauk (Jagiellonian University), Amanda Della Giustina (University of Ottawa), Kristina Sakers (Duke University)
Positive control: Rat astrocytes stained with DAKO anti-GFAP rabbit primary antibody, Alexa 546 conjugated secondary antibody, and DAPI for cell nuclei.
Caption: Positive control: Rat astrocytes stained with DAKO anti-GFAP rabbit primary antibody, Alexa 546 conjugated secondary antibody, and DAPI for cell nuclei.
Image attribution: Zuzanna Rauk (Jagiellonian University), Amanda Della Giustina (University of Ottawa), Kristina Sakers (Duke University)
Rat astrocytes stained with Addgene anti-GFAP mouse primary antibody (# 193445), Alexa 546 conjugated secondary antibody, and DAPI for cell nuclei.
Caption: Rat astrocytes stained with Addgene anti-GFAP mouse primary antibody (# 193445), Alexa 546 conjugated secondary antibody, and DAPI for cell nuclei.
Image attribution: Zuzanna Rauk (Jagiellonian University), Amanda Della Giustina (University of Ottawa), Kristina Sakers (Duke University)
Negative control for DAKO anti-rabbit GFP antibody: Rat astrocytes treated with blocking solution, PBST + NGS instead of primary antibody, and Alexa 546 conjugated secondary anti-rabbit antibody.
Caption: Negative control for DAKO anti-rabbit GFP antibody: Rat astrocytes treated with blocking solution, PBST + NGS instead of primary antibody, and Alexa 546 conjugated secondary anti-rabbit antibody.
Image attribution: Zuzanna Rauk (Jagiellonian University), Amanda Della Giustina (University of Ottawa), Kristina Sakers (Duke University)

This report is made available under the Creative Commons Attribution 4.0 International License.

Antibodies Used
Anti-GFAP [N206A/8R]
Addgene #193445
Goat anti-mouse IgG (H+L) cross-adsorbed secondary antibody, Alexa Fluor 546
Thermo Fisher #A-11003

Materials & Methods

Sample

Target Species
Rat
Cell / Tissue Type
Astrocytes/Cortex (Brain)

Buffer

Name
D-PBS

Fixative

Fixative
Formaldehyde/Paraformaldehyde

Permeabilization

Agent
Triton X-100
Concentration
0.4%
Time
30 min
Temperature
Room Temperature

Blocking

Agent
Normal Goat Serum (NGS)
Concentration
50%
Time
30 min
Temperature
Room Temperature

Primary Antibody

Name
Anti-GFAP [N206A/8R]
Source
Addgene
Catalog Number
193445
Host Species
Mouse
Target Antigen
GFAP
Concentration
1:1000
Time
overnight
Temperature
4 °C
View in Addgene Catalog

Secondary Antibody

Name
Goat anti-mouse IgG (H+L) cross-adsorbed secondary antibody, Alexa Fluor 546
Source
Thermo Fisher
Catalog Number
A-11003
Host Species
Goat
Concentration
1:500
Time
120 min

Additional Information

Additional Information

Rat primary astrocytes were stained with Anti-GFAP.

Exposure time: A546 800 ms; DAPI 500 ms

Results

Result
Pass (The antibody worked under these conditions)
Describe Results
This antibody gave a strong signal with low background.