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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 March 12, 2024)
Abstract: Experimental results for Anti-GFAP [N206A/8R] (Addgene Antibody 193445) in an immunocytochemistry assay with a rat sample.

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

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
Species: Mouse
Gene / Target: 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
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.