Overview

  • Product nameAnti-GRK5 antibody
    See all GRK5 primary antibodies
  • Description
    Rabbit polyclonal to GRK5
  • SpecificityGRK5 antibody detects endogenous levels of total GRK5 protein.
  • Tested applicationsSuitable for: IHC-Fr, WB, ELISA, ICC/IF, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide from internal of human GRK5.

  • Positive control
    • HUVEC, HepG2 and HeLa cell lysates

Properties

Applications

Our Abpromise guarantee covers the use of ab64943 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
IHC-Fr Use at an assay dependent concentration.
WB 1/500 - 1/1000. Detects a band of approximately 68 kDa (predicted molecular weight: 68 kDa).
ELISA 1/10000.
ICC/IF Use a concentration of 1 µg/ml.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

Target

Anti-GRK5 antibody images

  • ab64943 staining GRK5 in Rat H9C2 cells by ICC/IF (Immunocytochemistry/immunofluorescence). H9C2 cells were infected with Ad and were treated with GRK5 and PFA fro 24 hours. Cells were fixed with paraformaldehyde, permeabilized with 1% Triton X-100 and blocked with 5% serum for 1 hour at room temperature. Samples were incubated with primary antibody (1/200 in 5% Goat serum) for 2 hours. An undiluted Alexa Fluor® 488-conjugated Goat anti-rabbit IgG polyclonal was used as the secondary antibody.

    See Abreview

  • Anti-GRK5 antibody (ab64943) at 1/500 dilution + HEK293 whole cell lysate at 30 µg

    Secondary
    HRP-conjugated Goat anti-rabbit IgG polyclonal at 1/4000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 68 kDa
    Observed band size : 70 kDa (why is the actual band size different from the predicted?)


    Exposure time : 2 minutes

    This image is courtesy of an anonymous Abreview

    See Abreview

  • All lanes : Anti-GRK5 antibody (ab64943) at 1/500 dilution

    Lane 1 : HUVEC extract
    Lane 2 : HepG2 cell extract
    Lane 3 : HeLa cell extract
    Lane 4 : HeLa cell extract with immunising peptide


    Predicted band size : 68 kDa
    Observed band size : 68 kDa
    Additional bands at : 100 kDa (possible non-specific binding),40 kDa (possible non-specific binding).
  • ICC/IF image of ab64943 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 (ab64943, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 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.

  • IHC image of ab64943 staining in human breast carcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab64943, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

References for Anti-GRK5 antibody (ab64943)

ab64943 has not yet been referenced specifically in any publications.

Product Wall

Application Western blot
Loading amount 30 µg
Gel Running Conditions Reduced Denaturing (10% SDS PAGE)
Sample Mouse Tissue lysate - whole (Liver)
Specification Liver
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

Submitted Jul 23 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Frozen sections)
Sample Mouse Tissue sections (brain)
Specification brain
Fixative Paraformaldehyde
Permeabilization No
Blocking step Serum as blocking agent for 30 minute(s) · Concentration: 10% · Temperature: 20°C
Username

Mr. Daniel Havas

Verified customer

Submitted Sep 01 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Tissue lysate - whole (brain)
Loading amount 20 µg
Specification brain
Gel Running Conditions Reduced Denaturing (4-12% gradient)
Blocking step LiCor buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Mar 24 2011

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