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Western Blot for Anti-DYKDDDDK [IPI-4E11/M1]

Addgene #218106

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.dmta32
Abstract
Experimental results for Anti-DYKDDDDK [IPI-4E11/M1] (Addgene Antibody 218106) in a western blot assay with a human sample.
Citation
Western Blot for Anti-DYKDDDDK [IPI-4E11/M1]. Anderson Z, Zhu H, Riedel T, Moshinsky D, Meijers R 2024. Addgene Report, https://doi.org/10.57733/addgene.dmta32
Caption: Expi293 cells were transfected with Fab containing N-terminal (+DYKDDDDK-Fab-LC) or C-terminal (+Fab-LC-DYKDDDDK) DYKDDDDK tag or Fab alone (Neg. control). Cells were harvested, lysed, separated by SDS-PAGE, and immunoblotted against Anti-DYKDDDDK [IPI-4E11/M1]. This antibody performs best when the target is an N-terminal DYKDDDDK fusion.
Image attribution: Institute for Protein Innovation (IPI)

This report is made available under the Creative Commons Attribution 4.0 International License.

Antibodies Used
Anti-DYKDDDDK [IPI-4E11/M1]
Addgene #218106
IRDye 800CW Goat anti-Rabbit IgG
LICORbio #926-32211

Materials & Methods

Sample

Species
Human
Cell / Tissue Type
Expi293
Additional Information
Expi293 cells were transfected with Fab containing N-terminal (+DYKDDDDK-Fab-LC) or C-terminal (+Fab-LC-DYKDDDDK) DYKDDDDK 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-DYKDDDDK [IPI-4E11/M1]
Source
Addgene
Catalog Number
218106
Species
Rabbit
Gene / Target
DYKDDDDK
Concentration
1 µg/mL
Time
60 min
Temperature
Room Temperature
View in Addgene Catalog

Secondary Antibody

Name
IRDye 800CW Goat anti-Rabbit IgG
Source
LICORbio
Catalog Number
926-32211
Species
Goat
Concentration
1:20,000
Time
30 min
Temperature
Room Temperature

Additional Information

Additional Information
This antibody performs best when the target is an N-terminal DYKDDDDK fusion.
Detection Method
Fluorescence

Results

Result
Pass (The antibody worked under these conditions)
Describe Results
Strong signal for target