Anti-6X His tag®抗体[HIS.H8] (ab18184)

概述

性能

  • 形式Liquid
  • 存放说明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.
  • 存储溶液Preservative: 0.1% Sodium Azide
    Constituents: PBS, pH 7.2
  • Concentration information loading...
  • 纯度Ascites
  • 纯化说明Protein A affinity chromatography
  • 克隆单克隆
  • 克隆编号HIS.H8
  • 同种型IgG2b
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab18184 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
ICC/IF 1/200.
ELISA Use at an assay dependent dilution.
Dot blot Use at an assay dependent dilution.
WB 1/1000.
IP 1/200.
EMSA Use at an assay dependent dilution. PubMed: 20606007

靶标

  • 相关性The H-H-H-H-H-H motif is used as a tag on many recombinant proteins to facilitate purification. The antibody recognizes the His-tag fused to the amino- or carboxy- termini of targeted proteins in transfected or transformed cells.
  • 细胞定位Depends upon the localization of the parent protein tagged with hexahistidine.
  • 别名
    • 6 His epitope tag antibody
    • Hexa His tag antibody
    • HHHHHH epitope tag antibody
    • HHHHHH tag antibody
    • His tag antibody
    • Polyhistidine Tag antibody
    see all

Anti-6X His tag® antibody [HIS.H8] 图像

  • All lanes : Anti-6X His tag® antibody [HIS.H8] (ab18184) at 1/5000 dilution

    Lane 1 : ARPE-19 whole cell lysate - non-transfected
    Lane 2 : ARPE-19 whole cell lysate - transfected with rRab27a-His

    Lysates/proteins at 20 µg per lane.

    Secondary
    HRP-conjugated goat anti-mouse IgG polyclonal at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Exposure time : 5 minutes

    This image is courtesy of an Abreview submitted by Vladimir Milenkovic

    Blocked with 5% milk for 30 minutes at 25°C.

    Incubated with the primary antibody for 16 hours at 4°C in 1X PBS/Tween, 5% milk.

    See Abreview

  • ab18184 at a 1/1000 dilution staining 6X His tag in transfected human HEK293 cells by Immunocytochemistry/ Immunofluorescence incubated for 12 hours at 22°C. PFA fixed. Permeabilized using 0.1% Triton X-100 in PBS. Blocked using 3% BSA for 1 hour at 22°C. Secondary used at a 1/1000 dilution polyclonal Goat anti-mouse IgG (H+L) conjugated to Alexa Fluor 488 (green). DAPI staining (blue).

    See Abreview

  • All lanes : Anti-6X His tag® antibody [HIS.H8] (ab18184) at 1/1000 dilution

    Lane 1 : 6x His-tagged protein expressed in BL21 (pET15b) at 1 µg
    Lane 2 : 6x His-tagged protein expressed in BL21 (pET15b) at 0.1 µg

    Secondary
    IRDye® 800CW goat anti-mouse IgG (H+L) polyclonal at 1/5000 dilution

    Performed under reducing conditions.

    Exposure time : 5 minutes

    This image is courtesy of an Abreview submitted by Rachel Loh

    See Abreview

  • Immunoprecipitation of His/FLAG tagged PDCD5 using ab18184 at 1/100 dilution.

    Lane 1 : MW Ladder (from top 120, 85, 50, 34, 25, 19 kDa)
    Lanes 2-5 : Immunoprecipitation of His/FLAG tagged PDCD5 using ab18184 from mixture of ~50 ng purified His/FLAG tagged PDCD5 and 15 µL RRL in 300 µL buffer.
    Lanes 6-7 : Purified PDCD5 from E. coli with ~60-70 ng protein.

    The lowest band in each lane is ~ 20 kDa and is PDCD5. The upper bands in lanes 2-5 are the heavy chain (~50 kDa, runs as doublet) and light chain (~25 or 26 kDa).

    Western Blot was performed with a mouse anti-FLAG primary antibody at 1/2000 dilution. An IRDye®800CW-conjugated goat anti-mouse IgG was used as a secondary antibody.

    See Abreview

Anti-6X His tag® antibody [HIS.H8] (ab18184)参考文献

This product has been referenced in:
  • Tung H  et al. Baculovirus IE2 Stimulates the Expression of Heat Shock Proteins in Insect and Mammalian Cells to Facilitate Its Proper Functioning. PLoS One 11:e0148578 (2016). IP, ICC/IF . Read more (PubMed: 26863132) »
  • Jung B  et al. Pitchfork and Gprasp2 Target Smoothened to the Primary Cilium for Hedgehog Pathway Activation. PLoS One 11:e0149477 (2016). WB . Read more (PubMed: 26901434) »

See all 51 Publications for this product

Product Wall

Application Immunoprecipitation
Sample Human Cell lysate - whole cell (Embryonic fibrobalst cells)
Total protein in input 1e+006 cells
Treatment Influenza virus infection
Immuno-precipitation step Other - Protein G - Dynabeads
Specification Embryonic fibrobalst cells
Username

Abcam user community

Verified customer

提交于 Mar 25 2016

Abcam has not validated the combination of species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Spinal Cord (Dorsal Horn))
Antigen retrieval step None
Permeabilization Yes - 1% Triton, 0.25% Tween in 1XPBS
Specification Spinal Cord (Dorsal Horn)
Blocking step Mouse-on-Mouse Block (Vector Labs) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Feb 11 2016

Application Western blot
Sample Human Purified protein (S2 cell)
Gel Running Conditions Non-reduced Denaturing
Loading amount 0.001 µg
Specification S2 cell
Blocking step Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 May 27 2015

Application Western blot
Loading amount 1 µg
Gel Running Conditions Reduced Denaturing (12% hand cast gel)
Sample Escherichia coli Recombinant protein (6x His-tagged protein expressed in BL21 (pET15b))
Specification 6x His-tagged protein expressed in BL21 (pET15b)
Blocking step Odyssey Blocking Buffer (TBS) as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Miss. Rachel Loh

Verified customer

提交于 Jan 15 2015

Application IHC - Wholemount
Sample Xenopus tropicalis Embryo (whole embryo)
Specification whole embryo
Username

Abcam user community

Verified customer

提交于 Aug 04 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HeLa)
Loading amount 10 µg
Specification HeLa
Gel Running Conditions Reduced Denaturing (12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 May 01 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Recombinant protein (E. coli-purified human VCP-6xHis)
Loading amount 0.01 µg
Specification E. coli-purified human VCP-6xHis
Gel Running Conditions Reduced Denaturing (10% Acrylamide)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

提交于 Feb 21 2013

The main difference between ab18184 and ab5000 is the immunogen used to produce the antibodies. ab18184 was produced against a 6X His synthetic peptide, while ab5000 was raised against a recombinant protein fused to a 6X His tag. Thus, ab18184 was orig...

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Thank you for confirming these details and for your cooperation. The details provided enable us to closely monitor the quality of our products.

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this...

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - other (PDCD5 Purified from E Coli mixed with RRL)
Total protein in input 100 µg
Specification PDCD5 Purified from E Coli mixed with RRL
Immuno-precipitation step Protein A/G
Username

Dr. Chris Tracy

Verified customer

提交于 Dec 19 2012

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"