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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 May 28, 2024)
Abstract
Experimental results for Anti-DYKDDDDK [IPI-4E11/M1] (Addgene Antibody 218106) in a western blot assay with a human sample.
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