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

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.p0q07o
Abstract
Experimental results for Anti-GFAP [N206A/8R] (Addgene Antibody 193445) in an immunocytochemistry assay with a mouse sample.
Citation
Anti-GFAP [N206A/8R] in Immunocytochemistry (Mouse). Rauk Z, Della Giustina A, Sakers K, Pellegrino G 2024. Addgene Report, https://doi.org/10.57733/addgene.p0q07o
Negative control for DAKO anti-rabbit GFP antibody: Mouse 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: Mouse astrocytes treated with blocking solution, PBST + NGS instead of primary antibody, and Alexa 546 conjugated secondary anti-rabbit antibody.
Image attribution: CAJAL training programme
Negative control for Addgene antibody # 193445: Mouse 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: Mouse astrocytes treated with blocking solution, PBST + NGS instead of primary antibody, and Alexa 546 conjugated secondary anti-mouse antibody.
Image attribution: CAJAL training programme
Positive control: Mouse astrocytes stained with DAKO anti-GFAP rabbit primary antibody, Alexa 546 conjugated secondary antibody, and DAPI for cell nuclei.
Caption: Positive control: Mouse astrocytes stained with DAKO anti-GFAP rabbit primary antibody, Alexa 546 conjugated secondary antibody, and DAPI for cell nuclei.
Image attribution: CAJAL training programme
Mouse astrocytes stained with Addgene anti-GFAP mouse primary antibody (# 193445), Alexa 546 conjugated secondary antibody, and DAPI for cell nuclei.
Caption: Mouse astrocytes stained with Addgene anti-GFAP mouse primary antibody (# 193445), Alexa 546 conjugated secondary antibody, and DAPI for cell nuclei.
Image attribution: CAJAL training programme

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
Mouse
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

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
Concentration
1:500
Time
120 min

Additional Information

Additional Information

Mouse primary astrocytes were stained with Anti-GFAP.

Exposure time: A546 2000 ms; DAPI 50 ms

Results

Result
Fail (The antibody did not work under these conditions)
Describe Results
Antibody #193445 marks similar cells as the DAKO anti-GFAP rabbit primary antibody used in the positive control, but gave a weaker signal under the described experimental conditions.