Recombinant
RabMAb

Anti-KDEL antibody [EPR12668] - ER Marker (ab176333)

Overview

  • Product name
    Anti-KDEL antibody [EPR12668] - ER Marker
    See all KDEL primary antibodies
  • Description
    Rabbit monoclonal [EPR12668] to KDEL - ER Marker
  • 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

Images

  • All lanes : Anti-KDEL antibody [EPR12668] - ER Marker (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

  • 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.

  • 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. 

  • 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] - ER Marker (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.

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

    Lane 1 : HepG2 whole cell lysate
    Lane 2 : HeLa 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



    Obseved band size : 94,78,57,48 

    Blocking/Diluting buffer 5% NFDM/TBST

  • Immunohistochemical analysis of paraffin-embedded Human heart tissue labeling KDEL with ab176333 at 1/50 dilution

  • Immunohistochemical analysis of paraffin-embedded Human kidney tissue labeling KDEL with ab176333 at 1/50 dilution

  • Immunofluorescent staining of MCF7 cells labeling KDEL ER with ab176333 at 1/100 dilution (red). DAPI nuclear staining (blue).

  • Flow cytometric analysis of permeabilized HeLa cells labeling KDEL with ab176333 at 1/10 dilution (red) or a rabbit IgG (negative) (green).

References

ab176333 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

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
Username

Abcam user community

Verified customer

Submitted Jun 28 2018

Abcam guarantees this product to work in the species/application used in this Abreview.
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
Username

Michiel Krols

Verified customer

Submitted Apr 27 2016

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

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