Overview

  • Product name

    Anti-KDEL antibody [EPR12668]
    See all KDEL primary antibodies
  • Description

    Rabbit monoclonal [EPR12668] to KDEL
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide corresponding to KDEL.

  • Positive control

    • 293T, MCF-7, Raji, HepG2, HeLa, Jurkat, and K562 cell lysates. MCF7 and HeLa cells. Paraffin-embedded Human heart and kidney tissue.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.06% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR12668
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab176333 in the following tested applications.

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

Application Abreviews Notes
WB 1/1000 - 1/10000. Predicted molecular weight: 25 kDa.
IHC-P 1/50 - 1/350.

See IHC antigen retrieval protocols.

ICC/IF 1/100 - 1/250.
Flow Cyt 1/10 - 1/100.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • Relevance

    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.
  • Cellular localization

    Endoplasmic reticulum
  • Alternative names

    • KDEL antibody
    • Lys Asp Glu Leu antibody

Images

  • Immunocytochemistry/Immunofluorescence analysis of Jurkat cells labelling KDEL with purified ab176333 at 1/200. Cells were fixed with 4% Paraformaldehyde and permeabilized using 0.1% Triton X-100. ab150077, Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. Cells were co-stained with ab7291, a mouse anti-tubulin antibody (1/1000) using ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) as the secondary. Nuclei were counterstained with DAPI (blue).

    For negative control 1, ab150120 (Rabbit primary antibody) and anti-mouse secondary antibody were used and for negative control 2, ab7291 (Mouse primary antibody) and ab150077 (anti-rabbit secondary antibody) were used. 

  • All lanes : Anti-KDEL antibody [EPR12668] (ab176333) at 1/1000 dilution

    Lane 1 : 293T cell lysate
    Lane 2 : MCF7 cell lysate
    Lane 3 : Raji cell lysate
    Lane 4 : HepG2 cell lysate
    Lane 5 : HeLa cell lysate
    Lane 6 : Jurkat cell lysate
    Lane 7 : K562 cell lysate

    Lysates/proteins at 10 µg per lane.

    Predicted band size: 25 kDa

  • Lane 1 : Anti-P4HB antibody [EPR9498] (ab137119) at 1/5000 dilution
    Lanes 2 & 4 & 6 & 8 & 10 : Anti-KDEL antibody [EPR12668] (ab176333) at 1/5000 dilution
    Lane 3 : Anti-GRP78 BiP antibody [EPR4040(2)] (ab108613) at 1/5000 dilution
    Lane 5 : Anti-GRP94 antibody [EPR3988] (ab108606) at 1/5000 dilution
    Lane 7 : Anti-ERp57 antibody [EPR10678(B)] (ab154191) at 1/5000 dilution
    Lane 9 : Anti-PDIA6 antibody [EPR10132(B)] (ab154820) at 1/5000 dilution
    Lane 11 : Anti-LEPRE1/P3H1 antibody [EPR10193(B)] (ab154799) at 1/5000 dilution

    All lanes : HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate with 5% NFDM/TBST

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution

    Predicted band size: 25 kDa


    Exposure time: 3 minutes


    ab176333 detected several bands which have similar MWs with some proteins containing carboxyl terminal KDEL motifs, including P4HB, GRP78 BiP, GRP94, PDIA6 and LEPRE1 (PMID: 25683117, PMID: 19741001, PMID: 22079671, PMID: 28648146, PMID: 22615817).  It indicates that ab176333 might recognize these proteins.

  • Immunohistochemical analysis of paraffin-embedded human liver sections labelling KDEL with purified ab176333 at dilution of 1:350. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • Overlay histogram showing 4% paraformaldehyde fixed HeLa cells labelling KDEL (red) with purified ab176333 at dilution of 1/100. The secondary antibody used was Alexa Fluor® 488 goat-anti-rabbit IgG at dilution of 1/2000. A non-specific IgG antibody (rabbit monoclonal) was used as isotype control (black). The blue line shows cells without incubation with primary antibody and secondary antibody.

  • Immunohistochemical analysis of paraffin-embedded rat kidney sections labelling KDEL with purified ab176333 at dilution of 1:350. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

  • All lanes : Anti-KDEL antibody [EPR12668] (ab176333) at 1/10000 dilution

    Lane 1 : Raw264.7 whole cell lysate
    Lane 2 : C6 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/2000 dilution

    Predicted band size: 25 kDa



    Observed band size : 94,78,57,48

    Blocking/Diluting buffer 5% NFDM/TBST

  • Immunohistochemical analysis of paraffin-embedded mouse liver sections labelling KDEL with purified ab176333 at dilution of 1:350. The secondary antibody used was ab97051; a goat anti-rabbit IgG H&L (HRP) at dilution of 1/500. The sample was counterstained with hematoxylin. Antigen retrieval was performed using EDTA Buffer; pH 9.0. PBS was used instead of the primary antibody as the negative control and is shown in the inset.

References

This product has been referenced in:

See 1 Publication for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (Cancer Cell lines)
Gel Running Conditions
Reduced Denaturing (10)
Loading amount
25 µg
Specification
Cancer Cell lines
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 23°C

Abcam user community

Verified customer

Submitted Jun 28 2018

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (HeLa)
Permeabilization
Yes - 0.1% TritonX100 in PBS, 3mins
Specification
HeLa
Blocking step
2%BSA, 2% goat serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Fixative
Formaldehyde

Michiel Krols

Verified customer

Submitted Apr 27 2016

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