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Anti-SynCAM1 [L45/30R] in Western Blot (Rat)

Addgene #209587

Data Submitted By
James Trimmer
Lab Name
James Trimmer, UC Davis
Submission Date
June 17, 2024
Publication Date
July 30, 2024 (modified August 21, 2024)
Abstract
Experimental results for Anti-SynCAM1 [L45/30R] (Addgene #209587) in a western blot assay with a rat brain lysate. The staining of the Anti-SynCAM1 [L45/30R] recombinant antibody was compared to its parental hybridoma.
Caption: Crude rat membranes immunoblotted against Anti-SynCAM1 hybridoma-derived antibody from the Trimmer Lab at UC Davis (H) or purified recombinant antibody produced at Addgene (R).
Image attribution: James Trimmer, UC Davis

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

Antibodies Used
Anti-SynCAM1 [L45/30R]
Addgene #209587
Goat anti-mouse IgG2a
Jackson ImmunoResearch Labs #115-035-206

Materials & Methods

Sample

Species
Rat
Cell / Tissue Type
Brain
Additional Information
Rat brain lysates were boiled at 95 °C 5 min prior to loading.

Buffer

Name
TBS
pH
7.5

Blocking

Agent
Milk
Concentration
4% w/v
Time
45 min
Temperature
Room Temperature

Primary Antibody

Name
Anti-SynCAM1 [L45/30R]
Source
Addgene
Catalog Number
209587
Species
Mouse
Concentration
1 µg/mL
Time
45 min
Temperature
Room Temperature
View in Addgene Catalog

Secondary Antibody

Name
Goat anti-mouse IgG2a
Source
Jackson ImmunoResearch Labs
Catalog Number
115-035-206
Concentration
1:8333
Conjugate
HRP
Time
45 min
Temperature
Room Temperature

Additional Information

Additional Information
Total rat brain extract was reduced and separated by denaturing SDS-PAGE.
Protocol
Trimmer lab immunoblot labeling protocol
Detection Method
ECL
Predicted Molecular Weight
50 kDa

Results

Result
Pass (The antibody worked under these conditions)
Describe Results
Equivalent signal strength and labeling pattern between parental monoclonal antibody and recombinant antibody. The band is running higher than expected at ~75 kDa.