概述

  • 产品名称Anti-KDEL抗体[10C3]
    参阅全部 KDEL 一抗
  • 描述
    小鼠单克隆抗体[10C3] to KDEL
  • 特异性This antibody specifically recognizes proteins containing the SEKDEL sequence in mammals, birds, insects and plants. It recognises GPR78 and GPR94 with particular prominence. N.B Recent customer feedback has provided evidence that this antibody more specifically recognises the SEKDEL sequence rather than simply the KDEL sequence.
  • 经测试应用适用于: IHC-P, IHC-Fr, ICC/IF, WB, ICC, ELISA, IP, Flow Cytmore details
  • 种属反应性
    与反应: Mouse, Rat, Human, Chinese Hamster
    预测可用于: Bird, Plants, a wide range of other species, all Mammals
  • 免疫原

    Peptide sequence SEKDEL corresponding to amino acids 649-654 of rat Grp78 conjugated to KLH.

  • 阳性对照
    • HeLa cell lysate

性能

  • 形式Liquid
  • 存放说明Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
  • 存储溶液Preservative: None
    Constituents: 50% Glycerol, PBS, 0.1mM PMSF. pH 7.2
  • Concentration information loading...
  • 纯度Protein G purified
  • 克隆单克隆
  • 克隆编号10C3
  • 同种型IgG2a
  • 研究领域

应用

Our Abpromise guarantee covers the use of ab12223 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
IHC-P Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.
WB Use a concentration of 4 µg/ml. Predicted molecular weight: 78 , 94 kDa.
ICC Use a concentration of 5 µg/ml.
ELISA Use at an assay dependent concentration.
IP Use at an assay dependent concentration.
Flow Cyt Use 0.5µg for 106 cells.

ab170191 - Mouse monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

靶标

  • 相关性The sequence Lys-Asp-Glu-Leu (KDEL) or a closely related sequence, is present at the carboxy-terminus of soluble endoplasmic reticulum (ER) resident proteins and some membrane proteins. 78 and 94 kDa glucose regulated proteins (GRP 78) and GRP 94 respectively and protein disulfide isomerase (PDI) all share the C-terminal KDEL sequence. The presence of carboxy-terminal KDEL appears to be necessary for ER retention and appears to be sufficient to reduce the secretion of proteins from the ER. This retention is reported to be mediated by a KDEL receptor.
  • 细胞定位Endoplasmic reticulum
  • 数据库链接
  • 别名
    • KDEL antibody
    • Lys Asp Glu Leu antibody

Anti-KDEL antibody [10C3] 图像



  • Predicted band size : 78 , 94 kDa
    Three constructs were generated (denoted protein one to three). Proteins 1 and 2 end in SEKDEL as the C-terminal sequence. Protein two is an original construct which ends in GGKDEL. These were expressed and separated on a 12% SDS-PAGE, blotted to PVDF and blocked with 4% milk in TBS/0.1% tween. The replicates were then treated with either a specific antibody for the expressed protein (B), the KDEL antibody - ER Marker (ab2898) or KDEL antibody [10C3] - ER Marker (ab12223). A secondary antibody conjugated to alkaline phosphatase was used to detect the primary. The Coomassie stained gel indicates the location and expression levels of the three proteins. The specific antibody confirms the bands as the proteins of interest. KDEL antibody [10C3] - ER Marker (ab12223) recognizes proteins ending in the SEKDEL sequence and does not recognize the GGKDEL sequence as detected by KDEL antibody - ER Marker (ab2898).
  • ab12223 at 1/250 staining a mouse macropahge cell line by ICC/IF. The cells were paraformaldehyde fixed and blocked with BSA prior to incubation with the antibody for 24 hours. An Alexa-Fluor ® 488 conjugated donkey anti-mouse antibody was used as the secondary.

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  • ab12223 staining human Hella cells by ICC/IF. Cells were PFA fixed and permeabilized in 0.1% Triton X-100 for 10 minutes prior to blocking in 3% BSA for 15 minutes at room temperature. The primary antibody was diluted 1/50 and incubated with sample for 1 hour. A Alexa Fluor® 568 goat polyclonal to mouse IgG was used as the secondary, diluted 1/1000.

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  • ab12223 staining KDEL in mouse liver tissue section by Immunohistochemistry (Frozen sections). Tissue samples were fixed with formaldehyde and permeabilized with 0.2% Triton X-100 before blocking with 2% BSA for 30 minutes at 200C. The sample was incubated with primary antibody (1/200) for 9 hours at 40C. An Alexa Fluor® 555-conjugated Goat polyclonal to mouse IgG was used as secondary antibody at dilution at 1/200. DAPI was used to stain the cell nuclei (blue).

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  • ab12223 staining KDEL in Mouse kidney tissue by Immunohistochemistry (Formalin/ PFA-fixed paraffin-embedded tissue sections). The sections were Formaldehyde-fixed and subjected to Heat-mediated antigen retrieval in citric acid (pH 6.0) and permeabilized with 0.2% Triton X-100 prior to blocking with 5% BSA for 2 hours at 25°C. The primary antibody was diluted 1/250 in PBS and incubated with the sample for 9 hours at 4°C. An Alexa Fluor® 594-conjugated Goat anti-Mouse IgG polyclonal was used as the secondary antibody, diluted 1/500. A DAPI counterstain was used for nuclei.

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  • ICC/IF image of ab12223 stained HeLa cells. The cells were 4% PFA fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab12223, 10µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-mouse IgG - H&L, pre-adsorbed (ab96879) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

  • Overlay histogram showing HeLa cells stained with ab12223 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab12223, 0.5µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG2a [ICIGG2A] (ab91361, 1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

Anti-KDEL antibody [10C3] (ab12223)参考文献

This product has been referenced in:
  • Kondo S  et al. Secretory function in subplate neurons during cortical development. Front Neurosci 9:100 (2015). Read more (PubMed: 25859180) »
  • Cha-Molstad H  et al. Amino-terminal arginylation targets endoplasmic reticulum chaperone BiP for autophagy through p62 binding. Nat Cell Biol 17:917-29 (2015). Read more (PubMed: 26075355) »

See all 25 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Mouse Tissue sections (Kidney)
Specification Kidney
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate Buffer pH 6.0
Permeabilization Yes - 0.2% Triton X-100
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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Verified customer

提交于 Nov 25 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Tissue lysate - whole (293 FT cels)
Loading amount 50 µg
Specification 293 FT cels
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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Abcam user community

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提交于 Sep 01 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Rat Tissue sections (Testis)
Specification Testis
Fixative Formaldehyde
Permeabilization Yes - 0.1% Triton X-100
Blocking step BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Jul 16 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Rat Cell (Bone marrow cells)
Specification Bone marrow cells
Fixative Formaldehyde
Permeabilization Yes - 0.1% Triton X-100
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 May 20 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (293FT)
Specification 293FT
Fixative Formaldehyde
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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提交于 Jan 14 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Mouse Cell (Spleen)
Specification Spleen
Fixative Acetone
Permeabilization No
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
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提交于 Jan 04 2010

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (Liver)
Loading amount 100 µg
Specification Liver
Gel Running Conditions Non-reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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Verified customer

提交于 Nov 18 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (Liver)
Specification Liver
Fixative Formaldehyde
Permeabilization Yes - 0.2% Triton X-100
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 20°C
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Abcam user community

Verified customer

提交于 Nov 10 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (huh-7 cells)
Loading amount 15000 cells
Specification huh-7 cells
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
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提交于 Jul 27 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunocytochemistry/ Immunofluorescence
Sample Human Cell (HeLa)
Specification HeLa
Fixative Paraformaldehyde
Permeabilization Yes - 0.1% Triton X-100 (10 mins)
Blocking step BSA as blocking agent for 15 minute(s) · Concentration: 3% · Temperature: RT°C
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Dr. Dan Tattersall

Verified customer

提交于 Nov 05 2008

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