Recombinant
RabMAb

Recombinant Anti-ORP150 antibody [EPR5890] (ab124884)

Overview

  • Product name

    Anti-ORP150 antibody [EPR5890]
    See all ORP150 primary antibodies
  • Description

    Rabbit monoclonal [EPR5890] to ORP150
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, Flow Cytmore details
    Unsuitable for: ICC or IP
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human ORP150 aa 50-150. The exact sequence is proprietary.
    (Peptide available as ab156891)

  • Positive control

    • Human fetal liver, Rat liver, Rat pancreas, 293T, MCF7, MDA-MB-435S, SH-SY5Y and HeLa lysates; Human liver 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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab124884 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. Detects a band of approximately 150 kDa (predicted molecular weight: 111 kDa).Can be blocked with ORP150 peptide (ab156891).
IHC-P 1/250 - 1/500.
Flow Cyt 1/10 - 1/100.

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

 

  • Application notes
    Is unsuitable for ICC or IP.
  • Target

    • Function

      Has a pivotal role in cytoprotective cellular mechanisms triggered by oxygen deprivation. May play a role as a molecular chaperone and participate in protein folding.
    • Tissue specificity

      Highly expressed in tissues that contain well-developed endoplasmic reticulum and synthesize large amounts of secretory proteins. Highly expressed in liver and pancreas and lower expression in brain and kidney. Also expressed in macrophages within aortic atherosclerotic plaques, and in breast cancers.
    • Sequence similarities

      Belongs to the heat shock protein 70 family.
    • Cellular localization

      Endoplasmic reticulum lumen.
    • Information by UniProt
    • Database links

    • Alternative names

      • 150 kDa oxygen regulated protein antibody
      • 150 kDa oxygen-regulated protein antibody
      • 170 kDa glucose regulated protein antibody
      • 170 kDa glucose-regulated protein antibody
      • DKFZp686N08236 antibody
      • FLJ94899 antibody
      • FLJ97572 antibody
      • glucose regulated protein 170 antibody
      • Grp 170 antibody
      • GRP-170 antibody
      • Grp170 antibody
      • HSP 12A antibody
      • HSP12A antibody
      • Hyou1 antibody
      • HYOU1_HUMAN antibody
      • Hypoxia up regulated 1 antibody
      • hypoxia up-regulated 1 precursor antibody
      • Hypoxia up-regulated protein 1 antibody
      • Hypoxia upregulated 1 antibody
      • Orp 150 antibody
      • ORP-150 antibody
      • Orp150 antibody
      • oxygen regulated protein (150kD) antibody
      see all

    Images

    • All lanes : Anti-ORP150 antibody [EPR5890] (ab124884) at 1/1000 dilution

      Lane 1 : Human fetal liver lysate
      Lane 2 : Rat liver lysate
      Lane 3 : Rat pancreas lysate
      Lane 4 : 293T lysate
      Lane 5 : MCF7 lysate
      Lane 6 : MDA-MB-435S lysate
      Lane 7 : SH-SY5Y lysate
      Lane 8 : HeLa lysate

      Lysates/proteins at 10 µg/ml per lane.

      Secondary
      All lanes : HRP labelled goat anti-rabbit at 1/2000 dilution

      Predicted band size: 111 kDa

    • Overlay histogram showing HepG2 cells stained with ab124884 (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 (ab124884, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HepG2 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.
    • ab124884, at 1/250 dilution, staining ORP150 in paraffin-embedded Human liver tissue by Immunohistochemistry.
    • Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD

    References

    This product has been referenced in:

    • Balboa D  et al. Insulin mutations impair beta-cell development in a patient-derived iPSC model of neonatal diabetes. Elife 7:N/A (2018). Read more (PubMed: 30412052) »
    • Wang ZS  et al. Astragaloside IV attenuates proteinuria in streptozotocin-induced diabetic nephropathy via the inhibition of endoplasmic reticulum stress. BMC Nephrol 16:44 (2015). Read more (PubMed: 25886386) »
    See all 3 Publications for this product

    Customer reviews and Q&As

    1-2 of 2 Abreviews or Q&A

    Answer

    Thank you for taking the time to complete our questionnaire. I am sorry to hear you have had difficulty obtaining satisfactory results from this antibody.

    The details you have kindly provided will enable us to investigate this case for you and this is also helpful in our records for monitoring of quality.

    I can confirmthat this antibody was ordered in march and therefore iscovered by ourAbpromise guarantee.

    Reviewing this case, I would like to offer some suggestions to help optimise the results from ab124884.

    1.) All the suggestions form my last email:

    As a first troubleshooting tips I would like to suggest to review the protease inhibitors used since all extra bands are smaller than the target protein. They could be degradation products.

    I also suggest to run a no primary control to exclude the possibility that
    the secondary is producing the multiple extra bands.

    Since cell line that have been in culture for a long time can sometimes deliver unforeseen results, I suggest to use liver lysate as a positive control or a different cell line like HeLa or MCF7.

    2.) To achieve the most specific binding I recommend to incubate the primary antibody over night at 4C.

    3.) Some antibodies give better results when BSA is used as blocking agent. I can recommend to use 5% in the blocking buffer and to use 1% in the antibody solutions.


    I hope this information is helpful, thank you for your cooperation. Should the suggestions not improve the results, please do not hesitate to contact me again with the further requested details.

    Read More

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