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His-tag | 6xHis
英文名稱:His-tag | 6xHis總訪問:1123
國產(chǎn)/進口:進口半年訪問:10
產(chǎn)地/品牌:agrisera產(chǎn)品類別:植物試劑
規(guī)       格:AS111771 最后更新:2024-12-5
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product information
background His-Tag is a polyhistidine tag which consists of 6 histidine residues introduced on N- or C-terminus of the protein. The polyhistidine-tag can be used for recombinant protein detection using specific antibodies.
immunogen

KLH-conjugated synthetic peptide 6xHis

antibody format

mouse

monoclonal

IgG2b, clone HIS.H8 / EH158

lyophilized

quantity

100 µg

for reconstitution add 100 µl, of sterile water.

storage

store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. For storage at 4°C for several days to weeks, sodium azide to final 0.1 % can be added. Please, remember to spin tubes briefly prior to opening them to avoid any losses that might occur from lyophilized material adhering to the cap or sides of the tubes.

tested applications

western blot (WB), dot blot (Dot), ELISA (ELISA), immunoprecipiation (IP), immunlocalization (IL)

related products collection of antibodies to various tags
additional information

working dilution for ELISA, IL and IP needs to be determined experimentally.

application information
recommended dilution

1 : 1000 with standard ECL (WB)

expected | apparent MW

n.a.

confirmed reactivity

6xHis

predicted reactivity 6xHis
not reactive in

no confirmed exceptions from predicted reactivity are currently known

additional information

antibody is present in 10 mM PBS, pH 7.2

selected references

to be added when available, antibody available since July 2011.


application example

western blot on plant recombinant proteins using anti-His antibody

500 femtomoles of His-tagged proteins IsiA, NifH, PsbA, PsbB and PetC were loaded per gel well in Agrisera PEB extraction buffer. Proteins were separated on 4-12 % NuPAGE PAGE Bis-Tris polycacrylamide gel (Invitrogen) and blotted 1h to PVDF. Blots were blocked with for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in secondary antibody (goat, anti-mouse IgG horse radish peroxidase conjugated, from Agrisera AS11 1772) diluted to 1:25 000 in 2 % ECL Advance blocking reagent for 1h at RT with agitation. The blot was washed as above and developed for 5 min with ECL according to the manufacturers instructions. Exposure time was 5 seconds.

Apparent molecular weight of recombinant proteins: IsiA - 27 kda, NifH - 34 kDa, PsbA - 30-37 kDa, PsbB - 40 kDa, PetC - 23 kDa.

western blot using anti-His antibody from Agrisera


20 µl of media form Pichia Pastoris culter overexpressed His-Tegged proteins were separated on 12 % SDS-PAGE and blotted 1h to PVDF. Blots were blocked with 5% skim milk for 1h at room temperature (RT) with agitation. Blot was incubated in the primary antibody at a dilution of 1: 1 000 for 1h at RT with agitation. The antibody solution was decanted and the blot was rinsed briefly twice, then washed once for 15 min and 3 times for 5 min in TBS-T at RT with agitation. Blot was incubated in

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