Name:
Phone:
E-mail:
Address:






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)
His-tag | 3xHis (polyclonal)



DATA SHEET IN PDF

Qty: 
302
Buy 2 items of this product for 226.00 €/items
Buy 3 items of this product for 205.00 €/items
How to cite this product:
Product name, number (Agrisera, Sweden)

Data sheet Product citations Protocols Add review

Product Information

Immunogen

KLH-conjugated synthetic peptide 3xHis

Host Rabbit
Clonality Polyclonal
Purity Antigen affinity purified serum in PBS pH 7.4.
Format Lyophilized
Quantity 50 µg
Reconstitution For reconstitution add 50 µl, 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 Immunofluorescence (IF), Western blot (WB)
Recommended dilution 1 μg/mL (IF), 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. Luisa María Sandalio González, CSIC, Spain

Immunofluorescent localization using anti-3xHis rabbit polyclonal antibodies

HEK293 cells were transfected with the indicated plasmid (SARS-CoV-2 N myc tagged, PMID: 34799561, TBEV NS3-HA, PMID: 29321318, Rab5 mcherry, Addgene: 4920, GFP-HIS) using genejuice transfection reagent (EMD Millipore) according to the manufacturer’s instructions. After 24 hours of transfection, cells were fixed in 4% formaldehyde and permeabilized in PBS containing 0.5% Triton X-100 and 20 mM glycine. Then, cells were stained with the primary anti-3xHis tag antibodies at a concentration of 1 μg/mL for 1 hour at room temperature. Followed by three washes in PBS. Cells were then stained using secondary antibodies, donkey anti-rabbit Alexa555 (1:500, Thermo Fisher Scientific, a31572) in PBS containing 2% BSA for 1h at RT. Nuclei were stained using DAPI (1 μg/mL). Images were acquired using a Leica SP8 Laser Scanning Confocal Microscope with a 63x oil objective (Leica) and Leica Application Suit X software (LAS X, Leica).


Courtesy of Dr. Anna K Överby, Molecular Infection Medicine Sweden (MIMS),Section of Virology. Department of Clinical Microbiology Umeå University, Sweden

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

immunogen:

KLH-conjugated synthetic peptide 3xHis

Host: Rabbit
Clonality: Polyclonal
Purity: Antigen affinity purified serum in PBS pH 7.4.
Format: Lyophilized
Quantity: 50 µg
Reconstitution: For reconstitution add 50 µl, 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: Immunofluorescence (IF), Western blot (WB)
recommended dilution: 1 μg/mL (IF), 1: 1000 - 1 : 5000 (WB)
Expected | apparent MW: 3xHis tagged proteins
Confirmed reactivity: 3xHis tagged proteins
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. Luisa María Sandalio González, CSIC, Spain

Immunofluorescent localization using anti-3xHis rabbit polyclonal antibodies

HEK293 cells were transfected with the indicated plasmid (SARS-CoV-2 N myc tagged, PMID: 34799561, TBEV NS3-HA, PMID: 29321318, Rab5 mcherry, Addgene: 4920, GFP-HIS) using genejuice transfection reagent (EMD Millipore) according to the manufacturer’s instructions. After 24 hours of transfection, cells were fixed in 4% formaldehyde and permeabilized in PBS containing 0.5% Triton X-100 and 20 mM glycine. Then, cells were stained with the primary anti-3xHis tag antibodies at a concentration of 1 μg/mL for 1 hour at room temperature. Followed by three washes in PBS. Cells were then stained using secondary antibodies, donkey anti-rabbit Alexa555 (1:500, Thermo Fisher Scientific, a31572) in PBS containing 2% BSA for 1h at RT. Nuclei were stained using DAPI (1 μg/mL). Images were acquired using a Leica SP8 Laser Scanning Confocal Microscope with a 63x oil objective (Leica) and Leica Application Suit X software (LAS X, Leica).


Courtesy of Dr. Anna K Överby, Molecular Infection Medicine Sweden (MIMS),Section of Virology. Department of Clinical Microbiology Umeå University, Sweden

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.

Related products: His-tag | 3xHis (polyclonal)

AS16 ECL-S-N | low pico to mid femtogram and extreme low femtogram detection

Now available only...
From 26 €
AS20 4443 | Clonality: Polyclonal |  Host: Rabbit | Reactivity: Green Fluorescence Prote...
302 €
Buy 2 items of this product for 226.00 €/items
Buy 3 items of this product for 205.00 €/items
AS09 602 |  Clonality: Polyclonal | Host: Goat | Reactivity: Rabbit IgG (H&L)

10 ...
201 €
AS20 4457 | Clonality: Monoclonal | Host: Mouse | Reactivity: His-tagged proteins
367 €