Anti-Desmin [D7] ELISA against purified antigen
Addgene #201865
- Data Submitted By
- Meghan Rego
- Lab Name
- Addgene
- Submission Date
- May 22, 2024
- Publication Date
- October 09, 2024 (modified October 21, 2024)
- DOI
- https://doi.org/10.57733/addgene.oah5bb
- Funding
- National Institute of Health U24NS119916
- Abstract
- The binding specificity of purified recombinant Anti-Desmin [D7] was confirmed by testing in an ELISA against serial dilutions of purified human recombinant Desmin protein versus a negative control protein, Apolipoprotein L3. Anti-Desmin [D7] bound to Desmin protein in a dose-dependent manner but did not bind to Apolipoprotein L3.
- Citation
-
Anti-Desmin [D7] ELISA against purified antigen. Rego M 2024. Addgene Report, https://doi.org/10.57733/addgene.oah5bb
![The plate was coated with serial dilutions of human recombinant purified Desmin or Apolipoprotein L3 overnight, blocked, and incubated overnight with Anti-Desmin [D7]. Plates were incubated with HRP-linked anti-Human IgG secondary antibody for 2 hours, reacted with TMB substrate and the reaction stopped after 15 minutes. Absorbance was read at 450 nm and plotted against the micrograms of antigen loaded.](https://datahub-media.addgene.org/report-images/oah5bb/Screenshot_2024-05-22_at_11.58.07AM.png)
Image attribution: Meghan Rego
This report is made available under the Creative Commons Attribution 4.0 International License.
- Antibodies Used
-
Anti-Desmin [D7]
Addgene #201865 -
Goat anti-Human IgG (Gamma chain) Cross-Adsorbed Secondary Antibody, HRP
Thermo Fisher #62-8420
Materials & Methods
Sample
- Species
- Human
- Cell / Tissue Type
- Purified Recombinant Protein
- Additional Information
- Purified recombinant human Desmin (Origene #TP761590) and human APOL3 (Origene #TP761559) were obtained from Origene, as N-terminally GST-tagged and C-terminally His-tagged proteins, expressed in E. coli and purified.
ELISA Type
- Type
- Indirect
Coating
- Antigen / Antibody
- Desmin
- Concentration
- Dose ranged from 0 to 0.1 micrograms of protein
- Time
- Overnight
- Temperature
- 4 ˚C
Buffer
- Name
- PBS
- pH
- 7.4
Blocking
- Agent
- BSA
- Concentration
- 1% w/v in PBS pH 7.4
- Time
- 120 min
- Temperature
- Room Temperature
Primary Antibody
- Name
- Anti-Desmin [D7]
- Source
- Addgene
- Catalog Number
- 201865
- Species
- Human
- Gene / Target
- DES
- Concentration
- 0.25 µg/mL
- Time
- Overnight
- Temperature
- 4 ˚C
Secondary Antibody
- Name
- Goat anti-Human IgG (Gamma chain) Cross-Adsorbed Secondary Antibody, HRP
- Source
- Thermo Fisher
- Catalog Number
- 62-8420
- Species
- Goat
- Concentration
- 1:4000
- Conjugate
- HRP
- Time
- 120 min
- Temperature
- Room Temperature
Additional Information
- Additional Information
- Antibodies were diluted in 0.1% BSA in PBS.
- Protocol
- Addgene ELISA protocol
- Controls
- Positive control recombinant Desmin protein, Negative Control recombinat APOL3
- Wash Conditions
- 3x 1 min in PBS + 0.05% Tween-20
Results
- Result
- Pass (The antibody worked under these conditions)