Western Blot for Anti-His [IPI-HAP1/5.13.22]
Addgene #218109
- Data Submitted By
- Zachary Anderson, Haisun Zhu, Travis Riedel, Deborah Moshinsky and Rob Meijers
- Lab Name
- Institute for Protein Innovation (IPI)
- Submission Date
- April 29, 2024
- Publication Date
- May 24, 2024 (modified September 23, 2024)
- DOI
- https://doi.org/10.57733/addgene.gq7tmu
- Abstract
- Experimental results for Anti-His [IPI-HAP1/5.13.22] (Addgene Antibody 218109) in a western blot assay with a human sample.
- Citation
-
Western Blot for Anti-His [IPI-HAP1/5.13.22]. Anderson Z, Moshinsky D, Meijers R, Zhu H, Riedel T 2024. Addgene Report, https://doi.org/10.57733/addgene.gq7tmu
![Expi293 cells were transfected with Fab containing N-terminal (+His-Fab-LC) or C-terminal (+Fab-LC-His) His tag or Fab alone (Neg. control). Cells were harvested, lysed, separated by SDS-PAGE, and immunoblotted against Anti-His [IPI-HAP1/5.13.22]. This antibody performs best when the target is a C-terminal His fusion.](https://datahub-media.addgene.org/report-images/gq7tmu/WB_Anti-His_IPI-HAP1_5.13.22.png)
Image attribution: Institute for Protein Innovation (IPI)
This report is made available under the Creative Commons Attribution 4.0 International License.
- Antibodies Used
-
Anti-His [IPI-HAP1/5.13.22]
Addgene #218109 -
IRDye 800CW Goat anti-Rabbit IgG
LICORbio #926-32211
Materials & Methods
Sample
- Target Species
- Human
- Cell / Tissue Type
- Expi293
- Additional Information
- Expi293 cells were transfected with Fab containing N-terminal (+His-Fab-LC) or C-terminal (+Fab-LC-His) His tag or Fab alone (Neg. control).
Buffer
- Name
- PBS-T
Blocking
- Agent
- Milk
- Concentration
- 5%
- Time
- 30 min
- Temperature
- Room Temperature
Primary Antibody
- Name
- Anti-His [IPI-HAP1/5.13.22]
- Source
- Addgene
- Catalog Number
- 218109
- Host Species
- Rabbit
- Target Antigen
- His
- Concentration
- 1 µg/mL
- Time
- 60 min
- Temperature
- Room Temperature
Secondary Antibody
- Name
- IRDye 800CW Goat anti-Rabbit IgG
- Source
- LICORbio
- Catalog Number
- 926-32211
- Host Species
- Goat
- Concentration
- 1:20,000
- Time
- 30 min
- Temperature
- Room Temperature
Additional Information
- Additional Information
- This antibody performs best when the target is a C-terminal His fusion.
- Detection Method
- Fluorescence
Results
- Result
- Pass (The antibody worked under these conditions)
- Describe Results
- Strong signal for target