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宿主来源
Rabbit抗原名称
HA tag克隆号
S-4144抗体类型
Recombinant mAb抗体同种型
IgG反应种属 ?
Species Independent纯化方式
Protein A浓度
1 mg/ml标记
Unconjugated性状
Liquid缓冲体系
PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300
储存条件
12 months from date of receipt / reconstitution, -20 °C as supplied
应用
ICFCM
ChIP
ICC ?
WB
稀释度
应用 稀释度 推荐种属 WB 1:5000-1:50000 Species Independent ICC 1:1000 Species Independent ICFCM 1:1000 Species Independent ChIP 1:20-1:50 Species Independent
The HA tag (Hemagglutinin tag) is an epitope tag derived from the hemagglutinin (HA) protein of human influenza virus, typically consisting of a 9-amino acid sequence (YPYDVPDYA). Widely used in molecular biology and biochemistry due to its high immunogenicity and specificity, this tag is primarily employed for the detection, purification, and localization of proteins. By genetically fusing the HA tag to the N- or C-terminus of a target protein, high-affinity anti-HA antibodies can be utilized in experiments such as Western blot, co-immunoprecipitation (Co-IP), and immunofluorescence (IF) to effectively track the expression, subcellular localization, and interactions of the target protein. With its small size, high sensitivity, and minimal interference with protein structure, the HA tag has become an essential tool in protein research.
免疫印迹
WB result of HA Tag Recombinant Rabbit mAb
Primary antibody: HA Tag Recombinant Rabbit mAb at 1/10000 dilution
Lane 1: 293T transfected with empty vector whole cell lysate 20 µg
Lane 2: 293T transfected with HA-GFP fusion protein whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 28 kDa
Observed MW: 28 kDa
流式分析
Flow cytometric analysis of 4% PFA fixed 90% methanol permeabilized HA tag fusion protein transfected 293F cells (Human embryonic kidney epithelial cell) / (right panel) compared with untransfected 293F cells / (left panel) labeled with anti-HA tag antibody at 1/1000 (0.1 μg) dilution. Then cells were stained with anti-Myc Tag antibody at 1/2000 (0.1 μg) dilution (S0B0209) (Goat Anti - Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody). Flow cytometry and data analysis were performed using Agilent NovoCyte Quanteon and FlowJo™ software.
免疫细胞化学
ICC shows positive staining in HA tag fusion protein transfected 293F cells (top panel) and negative staining in vector-transfected 293F cells (below panel). Anti- HA Tag antibody was used at 1/1000 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 100% methanol and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
ChIP
Chromatin immunoprecipitation (ChIP) was performed on 293F cells were either untransfected (left panel) or transfected with an HA-tagged human H3 construct (right panel) cross - linked with 1% formaldehyde for 10 min, then chromatin was fragmented by sonication.
Parallel reactions used HA Tag Recombinant Rabbit mAb (S-4144) and Rabbit mAb IgG Isotype Control (SDT-R173) at 1:50 for immunoprecipitation.
Post - immunoprecipitation, both samples were washed, eluted, and cross - links reversed. Purified DNA was analyzed by qPCR.
qPCR showed the enrichment of RPL30, AFM and SAT-α
in HA Tag Recombinant Rabbit mAb (S-4144)-
immunoprecipitated sample.
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