Overview

  • Product name
  • Description
    Rabbit polyclonal to RIP140
  • Host species
    Rabbit
  • Specificity
    ab42126 recognises the C terminal domain of RIP140
  • Tested applications
    Suitable for: IHC-P, IHC (PFA fixed), IHC-Fr, IP, WB, ICC/IF, ChIPmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (Human) corresponding to the C terminal domain of RIP140. Peptide available as ab93493.

Properties

Applications

Our Abpromise guarantee covers the use of ab42126 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-P Use a concentration of 4 µg/ml.
IHC (PFA fixed) Use at an assay dependent concentration.
IHC-Fr Use at an assay dependent concentration.
IP 1/250.
WB 1/1000.
ICC/IF Use a concentration of 5 µg/ml.
ChIP Use at an assay dependent concentration. PubMed: 21360626

Target

  • Function
    Modulates transcriptional activation by steroid receptors such as NR3C1, NR3C2 and ESR1. Also modulates transcriptional repression by nuclear hormone receptors.
  • Domain
    Contains 9 Leu-Xaa-Xaa-Leu-Leu (LXXLL) motifs, which have different affinities for nuclear receptors. The C-terminal LTKTNPILYYMLQK motif is required for ligand-dependent interaction with RAAR and RXRB homodimers and heterodimers, for the corepressor activity, and for the formation of an HDAC3 complex with RARA/RXRB (By similarity). Contains at least four autonomous repression domains (RD1-4). RD1 functions via a histone deacetylase (HDAC)-independent mechanism, whereas RD2, RD3 and RD4 can function by HDAC-dependent or independent mechanisms, depending on cell type. RD2 is dependent on CTBP binding.
  • Post-translational
    modifications
    Acetylation regulates its nuclear translocation and corepressive activity (By similarity). Acetylation abolishes interaction with CTBP1. Phosphorylation enhances interaction with YWHAH.
  • Cellular localization
    Nucleus. Localized to discrete foci and redistributes to larger nuclear domains upon binding to ligand-bound NR3C1.
  • Information by UniProt
  • Database links
  • Alternative names
    • NRIP 1 antibody
    • NRIP1 antibody
    • NRIP1_HUMAN antibody
    • Nuclear factor RIP 140 antibody
    • Nuclear factor RIP140 antibody
    • Nuclear receptor interacting protein 1 antibody
    • Nuclear receptor-interacting protein 1 antibody
    • Receptor interacting protein 140 antibody
    • Receptor-interacting protein 140 antibody
    • RIP 140 antibody
    • RIP140 antibody
    see all

Images

  • ab42126 (4µg/ml) staining RIP140 in human colon using an automated system (DAKO Autostainer Plus). Using this protocol there is staining of the nuclei of the intestinal glands.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer EDTA pH 9.0 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.
  • ICC/IF image of ab42126 stained MCF7 cells. The cells were 4% formaldehyde 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 (ab42126, 5µg/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.

References

This product has been referenced in:
  • Lin YW  et al. Glyburide and retinoic acid synergize to promote wound healing by anti-inflammation and RIP140 degradation. Sci Rep 8:834 (2018). Read more (PubMed: 29339732) »
  • Yu XH  et al. Downregulation of RIP140 in triple-negative breast cancer inhibits the growth and proliferation of cancer cells. Oncol Lett 15:8784-8788 (2018). Read more (PubMed: 29844812) »
See all 23 Publications for this product

Customer reviews and Q&As

1-6 of 6 Abreviews or Q&A

Answer

Thank you for contacting us.

I can confirm, that to our knowledge, customised protocols are not required for the use of ab42126 - Anti-RIP140 antibody - ChIP Grade in IP. However, I can recommend to have a look at our standard protocol, provided on our Abcam Homepage here:

https://www.abcam.com/index.html?pageconfig=resource&rid=11385

https://www.abcam.com/index.html?pageconfig=resource&rid=11382

Furthermore, I would suggest to review the provided references provided for the use of this antibody on it's datasheet.

1. Ho PC et al. NF-?B-mediated degradation of the coactivator RIP140 regulates inflammatory responses and contributes to endotoxin tolerance. Nat Immunol 13:379-86 (2012).

"Cells were then collected and lysed in SDS lysis buffer"

2. http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3395431

I hope this information is helpful to you. Please do not hesitate to contact us if you have any problems or if you need any more advice or information.

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Answer

Thank you for your inquiry and your patience with this reply. I apologize that it took the testing laboratory a while to confirm how this antibody has been further tested. The lab confirmed that the antibody has been tested for labeling endogenous expression of this protein in cellular extracts, but since the protein is mainly present in nuclear extracts, the extraction is sometimes difficult.  They recommended a strong lysis buffer, such as RIPA buffer, or even doing a nuclear extraction (protocol linked below). RIPA buffer (Radio Immuno Precipitation Assay buffer) 150 mM sodium chloride 1.0% NP-40 or Triton X-100 0.5% sodium deoxycholate 0.1% SDS (sodium dodecyl sulphate) 50 mM Tris, pH 8.0 https://www.abcam.com/index.html?pageconfig=resource&rid=11408 Also since this is such a large protein, they recommended using an 8% gel. In addition, large proteins will tend to precipitate in the gel, hindering transfer. Adding SDS to a final concentration of 0.1% in the transfer buffer will discourage this. Methanol tends to remove SDS from proteins, so reducing the methanol percentage to 10% or less will also guard against precipitation. Plus, lowering methanol in the transfer buffer also promotes swelling of the gel, allowing large proteins to transfer more easily. But methanol is only necessary if using nitrocellulose. If using PVDF, methanol can be removed from the transfer buffer altogether, and is only needed to activate the PVDF before assembling the gel/membrane sandwich. Also you should choose to do a wet transfer overnight at 4oC instead of semi-dry transfer. Lastly the primary antibody was incubated 1/1000 overnight at 4C. If these suggestions do not improve your results and you have purchased the antibody in the last 6 months, I would be happy to replace or refund it for you. I hope this information helps and please let me know if you have any other questions.

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Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Blocking step
BSA as blocking agent for 3 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 20°C
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate
Sample
Human Tissue sections (Colon)
Specification
Colon
Permeabilization
No
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Apr 18 2014

Answer

Thank you for contacting us. This antibody was generate from an 18 aa peptide from the C terminal.
I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

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Answer

Thank you for contacting us.

This product's country of origin is the United States.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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

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