• Product name

  • Description

    Rabbit polyclonal to RNF144A
  • Host species

  • Tested applications

    Suitable for: IHC-P, ICC/IF, WB, ELISAmore details
  • Species reactivity

    Reacts with: Human, Rhesus monkey
    Predicted to work with: Mouse
  • Immunogen

    Synthetic peptide corresponding to Human RNF144A (internal sequence).

  • Positive control

    • extracts from COLO cells This antibody gave a positive result when used in the following formaldehyde fixed cell lines: HeLa. IHC-P: Human prostate FFPE tissue sections.
  • General notes

    Previously labelled as RNF144. 



Our Abpromise guarantee covers the use of ab75054 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 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use a concentration of 5 µg/ml.
WB 1/500 - 1/1000. Detects a band of approximately 30 kDa (predicted molecular weight: 33 kDa).
ELISA 1/40000.


  • Function

    E3 ubiquitin-protein ligase which accepts ubiquitin from E2 ubiquitin-conjugating enzymes UBE2L3 and UBE2L6 in the form of a thioester and then directly transfers the ubiquitin to targeted substrates.
  • Pathway

    Protein modification; protein ubiquitination.
  • Sequence similarities

    Belongs to the RBR family. RNF144 subfamily.
    Contains 1 IBR-type zinc finger.
    Contains 2 RING-type zinc fingers.
  • Cellular localization

  • Information by UniProt
  • Database links

  • Alternative names

    • KIAA0161 antibody
    • Probable E3 ubiquitin protein ligase RNF144A antibody
    • Probable E3 ubiquitin-protein ligase RNF144A antibody
    • R144A_HUMAN antibody
    • Ring finger protein 144 antibody
    • RING finger protein 144A antibody
    • RNF 144 antibody
    • RNF 144A antibody
    • RNF144 antibody
    • rnf144a antibody
    • UBCE7IP4 antibody
    • UbcM4 interacting protein 4 antibody
    • UbcM4-interacting protein 4 antibody
    • Ubiquitin conjugating enzyme 7 interacting protein 4 antibody
    • Ubiquitin-conjugating enzyme 7-interacting protein 4 antibody
    see all


  • All lanes : Anti-RNF144A antibody (ab75054) at 1/500 dilution

    Lane 1 : extracts from COLO cells
    Lane 2 : extracts from COLO cells with immunising peptide at 5 µg

    Lysates/proteins at 5 µg per lane.

    Predicted band size: 33 kDa
    Observed band size: 30 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 86 kDa. We are unsure as to the identity of these extra bands.

  • IHC image of RNF144A staining in human prostate formalin fixed paraffin embedded tissue section, performed on a Leica Bond 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 ab75054, 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.

  • ICC/IF image of ab75054 stained HepG2 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 ab75054 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) 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.


This product has been referenced in:

  • Chen X  et al. DHHC protein family targets different subsets of glioma stem cells in specific niches. J Exp Clin Cancer Res 38:25 (2019). Read more (PubMed: 30658672) »
  • Ho SR  et al. Regulation of RNF144A E3 Ubiquitin Ligase Activity by Self-association through Its Transmembrane Domain. J Biol Chem 290:23026-38 (2015). Read more (PubMed: 26216882) »
See all 4 Publications for this product

Customer reviews and Q&As

1-3 of 3 Abreviews or Q&A


Thanks for your reply with the images.

I was able to view the longer exposure image but the not the shorter exposure, I think the file was corrupted. However, I still got a good idea of the results with the long exposure. Here are my suggestions:

- Incubate the membrane at room temperature with less antibody (1:2,000 dilution). The background is high, so it is worth titrating the concentration to increase specific binding but reduced background.
- Block the membrane with at at least 1% milk, I wouldn't decrease this any further.
- Increase the washing time and perhaps the level of detergent? If you use Tween, you could switch to Triton (0.1%).
- Has the secondary antibody worked well with other primary antibodies? This will ensure the reactivity/sensitivity of the secondary has not changed.
- For sample preparation I would suggest using a RIPA based buffer or cellular fractionation in order to efficiently isolate the membrane

RIPA buffer
50 mM Tris HCl pH8
150 mM NaCl
1 % NP-40
0.5 % sodium deoxycholate
0.1 % SDS
+ protease inhibitors

Here is a link to our subcellular fractionation protocol on our website

- The lab confirmed that the antibody should be stored in single use aliquots.

I hope some of these improvements help and please let me know about your results.

Read More


Thank you for speaking with me today.

I have issued a free of charge replacement for ab75054 which is scheduled for dispatch today and should be with you tomorrow. Please aliquot the product into single use aliquots to prevent freeze thawing between each use. Once I receive the images from you I will consider what further changes to the protocol might help.

Looking forward to your reply with further information.

Read More

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