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Anti-HA [12CA5] - Chimeric in Western Blot (Human)

Addgene #198003

Data Submitted By: Meghan Rego
Lab Name: Addgene
Submission Date: February 06, 2024
Publication Date: February 06, 2024 (modified March 29, 2024)
Abstract: Experimental results for Anti-HA [12CA5] - Chimeric (Addgene Antibody 198003) in a western blot assay with a human sample.

HEK293 cells were left untransfected or transfected (+106410) with pCMV-P-RAN-RFP4. Cell culture supernatant containing the secreted protein was collected and probed with Anti-HA [12CA5] or Anti-6xHis [N144/14R]. — Caption: HEK293 cells were left untransfected or transfected (+106410) with pCMV-P-RAN-RFP4. Cell culture supernatant containing the secreted protein was collected and probed with Anti-HA [12CA5] or Anti-6xHis [N144/14R].
Image attribution: Addgene

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

Antibodies Used: Anti-HA [12CA5] - Chimeric
Addgene #198003; Goat anti-rabbit IgG Fc, HRP
Thermo Fisher #A16116

Materials & Methods

Sample

Species: Human
Cell / Tissue Type: HEK293
Total Amount: 30 µL

Buffer

Name: TBS-T
pH: 7.4

Blocking

Agent: Milk
Concentration: 5%
Time: 45 min
Temperature: Room Temperature

Primary Antibody

Name: Anti-HA [12CA5] - Chimeric
Source: Addgene
Catalog Number: 198003
Species: Rabbit
Concentration: 5 µg/mL
Time: 45 min
Temperature: Room Temperature

View in Addgene Catalog

Secondary Antibody

Name: Goat anti-rabbit IgG Fc, HRP
Source: Thermo Fisher
Catalog Number: A16116
Species: Rabbit
Concentration: unknown
Conjugate: HRP
Time: 45 min
Temperature: Room Temperature

Additional Information

Additional Information: Anti-6xHis used at 1 µg/mL and detected with goat anti-mouse IgG (H+L) HRP (Thermo Fisher, catalog number A16078)
Protocol: Addgene Western Blot protocol
Predicted Molecular Weight: 54 kDa

Results

Result: Pass (The antibody worked under these conditions)