Anti-6X His tag®抗体
参阅全部 6X His tag® 一抗
兔多克隆抗体to 6X His tag®
Rabbit
ELISA: Antibody specificity was verified by ELISA against peptide conjugated to BSA (HHHHHH-BSA). A 1:25,000 dilution of the antibody gave an OD=1.0 in a 15 minute reaction using HRP-conjugated Goat anti-Rabbit IgG at 1:20,000 and TMB as the substrate. Appropriate specificity controls were run.
适用于: WB, ICC/IF, ELISA, ChIPmore details
与反应: Species independent
Synthetic peptide corresponding to 6X His tag® conjugated to keyhole limpet haemocyanin.
ICC/IF: CHO cells transfected with 12 tags constructs (CHO-12 tags) and HEK293 expressing His tag. WB: HeLa transfected with His-tagged gene, Epitope Tag Positive Control whole cell lysates, purified 12-Tag and cell lysates of CHO-12 Tags.
This antibody recognizes both C-terminal and N-terminal tags.
Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077). Or search our wide range of secondary antibodies for use with your experiment.
HIS-TAG® is a trademark of EMD Biosciences, Inc.
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Liquid
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
pH: 7.20
Preservative: 0.1% Sodium azide
Constituent: PBS
浓度
100 µg 浓度为 1 mg/ml
Immunogen affinity purified
多克隆
IgG
Abpromise™承诺保证使用ab9108于以下的经测试应用
“应用说明”部分 下显示的仅为推荐的起始稀释度；实际最佳的稀释度/浓度应由使用者检定。
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB | (14) | Use a concentration of 1 µg/ml. |
| ICC/IF | (2) | 1/500 - 1/1000. |
| ELISA | (2) | 1/1000. |
| ChIP | Use at an assay dependent concentration. ChIP was performed with 25 ug chromatin, 5ug of antibody and 20 ul of Protein A/G beads. |
6 His epitope tag antibody
Hexa His tag antibody
HHHHHH epitope tag antibody
HHHHHH tag antibody
His tag antibody
Polyhistidine Tag antibody
Western blot - Anti-6X His tag® antibody (ab9108)
All lanes : Anti-6X His tag® antibody (ab9108) at 1 µg/ml
Lane 1 : Purified 12-Tag at 2 µg
Lane 2 : Purified 12-Tag at 0.2 µg
Lane 3 : The cell lysates of CHO-12 Tags at 2 µg
Lane 4 : The cell lysates of CHO at 2 µg
Secondary
All lanes : Goat anti-rabbit, HRP conjugated at 0.2 µg/ml
Lane 1 and 4: Molecular weight Standards.
Primary and secondary antibody incubation at RT for 1 hour.
Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody (ab9108)
Immunofluorescent analysis of 4% paraformaldehyde (PFA) -fixed, permeabilized with 0.1% Triton X-100 in CHO cells transfected with 12 tags constructs (CHO-12 tags) labelling 6xHIS with ab9108 at 1/1000 dilution (1µg/mL), followed by Donkey Anti-rabbit IgG (H&L), Alexa Fluor® 594 antibody at 1/500 dilution (4 µg/mL) (Red). Nucleus was counterstained with DAPI (Blue). Bottom panel images showed Parallel staining in parental CHO cell line.
Immunocytochemistry/ Immunofluorescence - Anti-6X His tag® antibody (ab9108)This image is courtesy of Vladimir Milenkovic
ab9108 staining CACNB3 with His tag expressed in HEK293 cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X-100 and blocked with 5% serum for 20 minutes at 25°C. Samples were incubated with primary antibody (1/500 in 1% goat serum, 0.1% TX100 and PBS) for 16 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody (1/750).
ChIP - Anti-6X His tag® antibody (ab9108)
A stably transfected 293T human cell line harbouring the GAL4 upstream activation sequence was transiently transfected with a Myc or His - tagged GAL4 DNA Binding Domain construct. 48 hours post transfection Chromatin was prepared according to the Abcam X-ChIP protocol. The ChIP was performed with 25 ug chromatin, 5ug of antibody and 20 ul of Protein A/G beads. A non-specific antibody was used as the negative control. The immunoprecipitated DNA was quantified by real time PCR (SYBR Green approach).
Western blot - Anti-6X His tag® antibody (ab9108)This image is courtesy of an anonymous Abreview
All lanes : Anti-6X His tag® antibody (ab9108) at 1/4000 dilution
Lane 1 : HeLa whole cell lysate - transfected with empty vector
Lane 2 : HeLa whole cell lysate - transfected with His-tagged gene
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP-conjugated goat anti-rabbit IgG polyclonal at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Observed band size: 35 kDawhy is the actual band size different from the predicted?
Western blot - Anti-6X His tag® antibody (ab9108)
Use of ab9108 in western blotting at 1:1000 against Epitope Tag Positive Control lysate (ab4524).
Lane 1: 2 µl lysate
Lane 2: 1 µl lysate
Lane 3: 0.5 µl lysate
Lane 4: 0.25 µl lysate Use of ab9108 in western blotting at 1:1000 against E.coli Positive Control (Escherichia coli ) Whole Cell Lysate - expressing 11 different epitope tag (ab5395). Lane 1: 2 µl lysate Lane 2: 1 µl lysate
