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His-tag | 3xHis (polyclonal)

AS20 4441   | Clonality: Polyclonal  |  Host: Rabbit  | Reactivity: 3xHis-tag
 

His-tag | 3xHis (polyclonal) in the group Tag Antibodies / His (3x, 6x) at Agrisera AB (Antibodies for research) (AS20 4441)



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Product Information

Immunogen

KLH-conjugated synthetic peptide 3xHis

Host Rabbit
Clonality Polyclonal
Purity Immunogen affinity purified serum in PBS pH 7.4.
Format Lyophilized
Quantity 50 ĩg
Reconstitution For reconstitution add 50 ĩl, of sterile water
Storage Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
Tested applications Western blot (WB)
Recommended dilution 1: 1000 - 1 : 5000 (WB)
Expected | apparent MW 3xHis tagged proteins

Reactivity

Confirmed reactivity 3xHis tagged proteins

Application examples

Application examples Western blot using polyclonal anti-3xHIS antibodies

Samples:
40 µg of E. coli supernatant in control conditions at 3h (1)
40 µg of E. coli pellet in control conditions at 3h (2)
40 µg of E. coli supernatant induced at 3h (3)
40 µg of E. coli pellet induced at 3h (4)

40 µg of total protein from Escherichia coli expressing pGEX-PEX11a inducible by IPTG, extracted with HEPES 50 mM pH 7.6, 300 mM NaCl, 5% glycerol, 20 mM imidazole, 1 mM MgCl2, 1 x protease inhibitor; prepared in 0.063 M Tris-HCl buffer, pH 6.8, containing 2% sodium dodecyl sulfate (SDS; w/v), 10 % glycerol (v/v), 0.006 % bromophenol blue (w/v) and 10 mM DTT; and denaturated at 95 ºC for 5 min. Proteins were separated on 12 % SDS-PAGE and blotted 1 h to PVDF using semi-dry transfer. Blots were blocked with 3% (w/v) skimmed milk powder diluted in TBS-T O/N at 4 ºC with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5.000 for 1 h at RT with agitation in TBS-T + 3% (w/v) of skimmed milk powder. The antibody solution was decanted and the blot was washed 3 times for 10 min in TBS-T + 3 % (w/v) of skimmed milk powder at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, Product No: AS09 602) diluted to 1:10.000 in TBS-T + 3 % (w/v) of skimmed milk powder for 1 h at RT with agitation. The blot was washed as above but with TBS and developed for 5 min with chemiluminescent detection reagent following manufacture's recommendations. Exposure time was 30 seconds.

Courtesy Dr. uisa María Sandalio González, CSIC, Spain

Additional information

Related products

Background

Background His-Tag is a polyhistidine tag which consists of 3 or 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 and it is not conjugated to any dye or enzyme.

Product citations

background: His-Tag is a polyhistidine tag which consists of 3 or 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 and it is not conjugated to any dye or enzyme.
Picture (footer): Western blot using polyclonal anti-3xHIS antibodies

Samples:
40 µg of E. coli supernatant in control conditions at 3h (1)
40 µg of E. coli pellet in control conditions at 3h (2)
40 µg of E. coli supernatant induced at 3h (3)
40 µg of E. coli pellet induced at 3h (4)

40 µg of total protein from Escherichia coli expressing pGEX-PEX11a inducible by IPTG, extracted with HEPES 50 mM pH 7.6, 300 mM NaCl, 5% glycerol, 20 mM imidazole, 1 mM MgCl2, 1 x protease inhibitor; prepared in 0.063 M Tris-HCl buffer, pH 6.8, containing 2% sodium dodecyl sulfate (SDS; w/v), 10 % glycerol (v/v), 0.006 % bromophenol blue (w/v) and 10 mM DTT; and denaturated at 95 ºC for 5 min. Proteins were separated on 12 % SDS-PAGE and blotted 1 h to PVDF using semi-dry transfer. Blots were blocked with 3% (w/v) skimmed milk powder diluted in TBS-T O/N at 4 ºC with agitation. Blot was incubated in the primary antibody at a dilution of 1: 5.000 for 1 h at RT with agitation in TBS-T + 3% (w/v) of skimmed milk powder. The antibody solution was decanted and the blot was washed 3 times for 10 min in TBS-T + 3 % (w/v) of skimmed milk powder at RT with agitation. Blot was incubated in secondary antibody (anti-rabbit IgG horse radish peroxidase conjugated, from Agrisera, Product No: AS09 602) diluted to 1:10.000 in TBS-T + 3 % (w/v) of skimmed milk powder for 1 h at RT with agitation. The blot was washed as above but with TBS and developed for 5 min with chemiluminescent detection reagent following manufacture's recommendations. Exposure time was 30 seconds.

Courtesy Dr. uisa María Sandalio González, CSIC, Spain

calculated | apparent molecular mass [kDa]: 3xHis tagged proteins
Clonality: Polyclonal
Format: Lyophilized
Host: Rabbit
immunogen:

KLH-conjugated synthetic peptide 3xHis

Purity: Immunogen affinity purified serum in PBS pH 7.4.
Quantity: 50 ĩg
recommended dilution: 1: 1000 - 1 : 5000 (WB)
Reconstitution: For reconstitution add 50 ĩl, of sterile water
storage: Store lyophilized/reconstituted at -20°C; once reconstituted make aliquots to avoid repeated freeze-thaw cycles. Please remember to spin the tubes briefly prior to opening them to avoid any losses that might occur from material adhering to the cap or sides of the tube.
tested applications: Western blot (WB)
Confirmed reactivity: 3xHis tagged proteins

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