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Western blot analysis of Hemagglutinin was performed by loading 25ug of 293t cells transfected with 12Tag onto an SDS polyacrylamide gel. Proteins were transferred to a PVDF membrane and blocked at 4°C overnight. The membrane was probed with a HA Epitope Tag polyclonal antibody (Product # PA1-985) at a dilution of 1:1000 overnight at 4°C, washed in TBST, and probed with an HRP-conjugated secondary antibody for 1 hr at room temperature in the dark. Chemiluminescent detection was performed using Pierce ECL Plus Western Blotting Substrate (Product # 32132). Results show a band at ~45 kDa.
|Tested species reactivity||Tag|
|Published species reactivity||Not Applicable|
|Host / Isotype||Rabbit / IgG|
|Immunogen||Synthetic peptide corresponding to the residues Y P Y D V P D Y A of the HA epitope of influenza hemagglutinin.|
|Purification||Antigen affinity chromatography|
|Storage buffer||PBS with 1mg/ml BSA|
|Contains||0.05% sodium azide|
|Storage Conditions||-20° C, Avoid Freeze/Thaw Cycles|
|Tested Applications||Dilution *|
|Western Blot (WB)||1:500-1:5000|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
|Western Blot (WB)||See 1 publications below|
PA1-985 detects HA tagged proteins. This antibody is specific for the HA tag and does not detect other proteins.
PA1-985 has been successfully used in Western blot procedures. By Western blot, this antibody detects the presence of HA-tagged proteins.
The PA1-985 immunogen is a synthetic peptide corresponding to the residues Y P Y D V P D Y A of the HA epitope of influenza hemagglutinin.
Epitope tagging is a powerful and versatile strategy for detecting and purifying proteins expressed by cloned genes. To utilize this feature, protein expression vectors are typically engineered with a nucleotide sequence that encodes the epitope tag. The gene of interest is cloned in-frame relative to the tag and, upon expression, the protein of interest is synthesized as a fusion protein with the epitope tag. Fusion protein detection and/or purification is mediated by highly specific antibodies to the engineered protein, thus eliminating the need for antibodies to proteins from each newly cloned gene. Commonly used epitope tags include glutathione-S-transferase (GST), c-myc, 6-histidine (6X-His), FLAG® green fluorescent protein (GFP), maltose binding protein (MBP), influenza A virus haemagglutinin (HA), b-galactosidase, and GAL4.
FLAG® and anti-FLAG® are registered trademarks of Sigma-Aldrich Biotechnology Co.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Identification of PAN2 by quantitative proteomics as a leucine-rich repeat-receptor-like kinase acting upstream of PAN1 to polarize cell division in maize.
PA1-985 was used in western blot to study the roles of PAN1 and PAN2 in the polarization of maize cell division
|Zhang X,Facette M,Humphries JA,Shen Z,Park Y,Sutimantanapi D,Sylvester AW,Briggs SP,Smith LG||The Plant cell (24:4577)||2012|
HA; HA Epitope Tag; HA-tag; human influenza hemagglutinin