Overview

  • Product nameAnti-p23 antibody [JJ3]
    See all p23 primary antibodies
  • Description
    Mouse monoclonal [JJ3] to p23
  • Tested applicationsSuitable for: ICC/IF, Inhibition Assay, Flow Cyt, IHC-P, WB, IPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Chicken, Guinea pig, Human, Xenopus laevis
    Predicted to work with: Hamster, Monkey, Non Human Primates
  • Immunogen

    Recombinant full length protein corresponding to Human p23.

Properties

  • FormLiquid
  • Storage instructionsShipped 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.
  • Storage bufferPreservative: 0.05% Sodium azide
    Constituent: PBS
  • Concentration information loading...
  • PurityAscites
  • Primary antibody notesSteroid receptors are ligand-dependent intracellular proteins that stimulate transcription of specific genes by binding to specific DNA sequences following activation by the appropriate hormone. Prior to activation, steroid receptors associate with a number of different proteins in both a stable and transient fashion. Steroid receptor complex proteins include heat shock proteins (HSP 70 and HSP 90), immunosuppressant binding proteins called immunophilins (the FK 506 binding proteins, FKBP 52 & FKBP 54 and the cyclosporin binding protein, CyP-40) and at least three other proteins termed p23, p60 and p48. p23 along with HSP 70, HSP 90 and p60, combine with progesterone receptor (PR) as members of a transient intermediate complex. Cloned human p23 encodes a protein of 160 amino acids that is highly conserved between species and shows no homology to previously identified proteins. p23 is a highly acidic phosphoprotein with an aspartic acid-rich C-terminal domain and multiple potential phosphorylation sites. In vitro studies have suggested that p23 binds to HSP90 and is necessary for the binding of HSP 90 and CyP-40 to PR. While neither its exact function nor mechanism of action have been identified, p23 appears to be an important factor in PR function.
  • ClonalityMonoclonal
  • Clone numberJJ3
  • IsotypeIgG1
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab2814 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
ICC/IF 1/200 - 1/2000.
Inhibition Assay Use at an assay dependent concentration.
Flow Cyt 1/100.
IHC-P Use at an assay dependent concentration.
WB 1/1000. Predicted molecular weight: 18 kDa. This antibody detects a 23 kDa protein representing p23 from rabbit reticulocyte lysate.
IP Use at an assay dependent concentration. This antibody can be used to immunoprecipitate both free and complexed p23.

Target

Anti-p23 antibody [JJ3] images

  • Immunocytochemistry/Immunofluorescence analysis of p23 shows staining in C6 cells. p23 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2814 (1:500) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse IgG secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of p23 shows staining in HeLa cells. p23 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2814 (1:500) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse IgG secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/Immunofluorescence analysis of p23 shows staining in MCF-7 cells. p23 staining (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2814 (1:500) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated goat anti-mouse IgG secondary antibody. Images were taken at 60X magnification.

  • Flow cytometry analysis of p23 showing positive staining in the cytoplasm of 3T3 cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/ml, fixed with 2% paraformaldehyde, washed with PBS, and incubated with ab2814 (0.5 ug/test) for 60 min at room temperature. Cells were then blocked in a solution of 2% BSA-PBS for 30 min at room temperature, incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody, and re-suspended in PBS for FACS analysis.

  • Flow cytometry analysis of p23 showing positive staining in the cytoplasm of Hela cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/ml, fixed with 2% paraformaldehyde, washed with PBS, and incubated with ab2814 (0.25 ug/test) for 60 min at room temperature. Cells were then blocked in a solution of 2% BSA-PBS for 30 min at room temperature, incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody, and re-suspended in PBS for FACS analysis.

  • Flow cytometry analysis of p23 showing positive staining in the cytoplasm of Jurkat cells compared to an isotype control (blue). Cells were harvested, adjusted to a concentration of 1-5x10^6 cells/ml, fixed with 2% paraformaldehyde, washed with PBS, and incubated with ab2814 (0.25 ug/test) for 60 min at room temperature. Cells were then blocked in a solution of 2% BSA-PBS for 30 min at room temperature, incubated for 40 min at room temperature in the dark using a Dylight 488-conjugated goat anti-mouse IgG (H+L) secondary antibody, and re-suspended in PBS for FACS analysis.

  • All lanes : Anti-p23 antibody [JJ3] (ab2814) at 1/1000 dilution

    Lane 1 : HeLa cell lysate
    Lane 2 : HepG2 cell lysate
    Lane 3 : Mouse spleen cell lysate

    Lysates/proteins at 25 µg per lane.


    Predicted band size : 18 kDa
    Observed band size : 23 kDa (why is the actual band size different from the predicted?)
  • ab2814 immunoprecipitating p23 from rat PC12 cells (20ug whole cell lysate). A protein G matrix was used with the antibody at a concentration of 4ug/mg of lysate. ab2814 was then used in the western blot stage of the experiment at a dilution of 1/1000.

    Lane 1: p23 IP

    Lane 2: Control mouse IgG IP

    Lane 3: Input (20%)

    See Abreview

  • ab2814 at 1/200 staining human anal cancer tissue sections by IHC-P. The tissue was paraformaldehyde fixed and a heat mediated antigen retrieval step was performed in citrate buffer, before the tissue was blocked and incubated with the antibody for 45 minutes. An HRP conjugated rabbit anti-mouse antibody was used as the secondary.

    See Abreview

  • Overlay histogram showing HeLA cells stained with ab2814 (red line). The cells were fixed with 4% paraformaldehyde (10 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 (ab2814, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 100% methanol (5 min)/permeabilized in 0.1% PBS-Tween used under the same conditions.

  • ICC/IF image of ab2814 stained MCF7 cells. The cells were 100% methanol fixed (5 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 (ab2814, 5µg/ml) overnight at +4°C. The secondary antibody (green) was ab96879, DyLight® 488 goat anti-mouse IgG (H+L) 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.


  • developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 18 kDa


    Exposure time : 90 seconds
  • Immunohistochemistry was performed on normal biopsies of deparaffinized Human tonsil tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:1000 with a mouse monoclonal antibody recognizing p23 ab2814 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on cancer biopsies of deparaffinized Human breast carcinoma tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:1000 with a mouse monoclonal antibody recognizing p23 ab2814 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

  • Immunohistochemistry was performed on normal biopsies of deparaffinized Human testis tissue. To expose target proteins heat induced antigen retrieval was performed using 10mM sodium citrate (pH6.0) buffer microwaved for 8-15 minutes. Following antigen retrieval tissues were blocked in 3% BSA-PBS for 30 minutes at room temperature. Tissues were then probed at a dilution of 1:1000 with a mouse monoclonal antibody recognizing p23 ab2814 or without primary antibody (negative control) overnight at 4°C in a humidified chamber. Tissues were washed extensively with PBST and endogenous peroxidase activity was quenched with a peroxidase suppressor. Detection was performed using a biotin-conjugated secondary antibody and SA-HRP followed by colorimetric detection using DAB. Tissues were counterstained with hematoxylin and prepped for mounting.

References for Anti-p23 antibody [JJ3] (ab2814)

This product has been referenced in:
  • Cano LQ  et al. The co-chaperone p23 promotes prostate cancer motility and metastasis. Mol Oncol 9:295-308 (2015). WB, IHC ; Human . Read more (PubMed: 25241147) »
  • El-Kasaby A  et al. A cytosolic relay of heat shock proteins HSP70-1A and HSP90ß monitors the folding trajectory of the serotonin transporter. J Biol Chem 289:28987-9000 (2014). Read more (PubMed: 25202009) »

See all 15 Publications for this product

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Unfortunately, ab2814 is an unpurified antibody (provided as ascites). We can only provide concentrations for purified antibodies as unpurified antibody preparations vary significantly in specific antibody concentration.

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Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Hek293T)
Loading amount 30 µg
Specification Hek293T
Gel Running Conditions Reduced Denaturing (12%)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
Username

Dr. Mirjam Eckert

Verified customer

Submitted Oct 29 2009

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample Human Tissue sections (lung carcinoma)
Specification lung carcinoma
Fixative Formaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate pH 6.0
Permeabilization No
Blocking step Serum as blocking agent for 10 minute(s) · Concentration: 1.5%
Username

Abcam user community

Verified customer

Submitted Sep 24 2008

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunoprecipitation
Sample Human Cell lysate - whole cell (293T)
Total protein in input 500 µg
Specification 293T
Immuno-precipitation step Protein G
Username

Abcam user community

Verified customer

Submitted Oct 30 2007

Application Immunoprecipitation
Sample Rat Cell lysate - whole cell (PC12)
Total protein in input 500 µg
Specification PC12
Immuno-precipitation step Protein G
Username

Abcam user community

Verified customer

Submitted Oct 30 2007

Application Western blot
Sample African Green Monkey Cell lysate - whole cell (Cos-7 cell line)
Loading amount 30 µg
Specification Cos-7 cell line
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Sep 17 2007

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Mouse Cell lysate - whole cell (3T3 cell line)
Loading amount 30 µg
Specification 3T3 cell line
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Sep 17 2007

Application Western blot
Sample Hamster Cell lysate - whole cell (CHO cell line)
Loading amount 30 µg
Specification CHO cell line
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Sep 17 2007

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 (small intestine)
Specification small intestine
Fixative Paraformaldehyde
Antigen retrieval step Heat mediated - Buffer/Enzyme Used: Citrate pH 6.0
Permeabilization No
Blocking step Serum as blocking agent for 20 minute(s) · Concentration: 1% · Temperature: 22°C
Username

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Verified customer

Submitted Jul 31 2007

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