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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.