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Rabbit抗原名称
HA tag免疫原
Synthetic Peptide克隆号
SDT-113-76抗体类型
Rabbit mAb应用
ICFCM, ChIP, ICC, WB反应种属 ?
Species Independent纯化方式
Protein A浓度
0.5 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 | 稀释度 |
|---|---|
| WB | 1:5000-1:50000 |
| ICFCM | 1:500 |
| ICC | 1:500 |
| ChIP | 1:20-1:50 |
HA-tag is derivative of self -influenza hemaginin, which is widely used as general table tags in the expression carrier.
免疫印迹
WB result of HA tag Recombinant Rabbit mAb (SDT-113-76)
Lysate: GST Protein, HA tag 20 ng
Primary antibody:
Lane 1: HA tag Recombinant Rabbit mAb at 1/5000 dilution
Lane 2: HA tag Recombinant Rabbit mAb at 1/10000 dilution
Lane 3: HA tag Recombinant Rabbit mAb at 1/50000 dilution
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
WB result of HA Rabbit mAb
Primary antibody: HA Rabbit mAb at 1/5000 dilution
Lane 1: 293F whole cell lysate 20 µg
Lane 2: 293F transfected with HA-tag-fusion protein whole cell lysate 20 µg
Secondary antibody: Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 32 kDa
Observed MW: 32 kDa
流式分析
Flow cytometric analysis of 4% PFA fixed 0.1% Tween-20 permeabilized Histone H3, GST Tag, HA Tag, Myc tag transfected 293T (Human embryonic kidney epithelial cell) labelling HA tag antibody at 1/500 (0.1 μg) dilution/ (right panel) compared with a Rabbit IgG Isotype Control / (left panel). Goat Anti-Rabbit IgG Alexa Fluor® 488 was used as the secondary antibody. Then cells were stained with Myc - Alexa Fluor® 647 separately.
免疫细胞化学
ICC shows positive staining in Histone H3-HA Tag transfected 293T cells (left panel) and negative staining in vector-transfected 293T cells (right panel). Anti-HA tag antibody was used at 1/500 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% ice-cold 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 293T 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 (SDT-113-76), Histone H3 Recombinant Rabbit mAb (SDT-266-44) 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 (%input: immunoprecipitated DNA/input DNA) showed the enrichment of RPL30, MYOD1 and SAT-2 in HA tag Recombinant Rabbit mAb (SDT-113-76)-immunoprecipitated sample.
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