Overview

  • Product name
  • Description
    Rabbit polyclonal to RFP2
  • Tested applications
    Suitable for: WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
    Predicted to work with: Chimpanzee
  • Immunogen

    Synthetic peptide derived from residues 50 - 150 of Human RFP2.

    .

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: HeLa; A431; HEk293 as well as HeLa Nuclear lysate.

Properties

Applications

Our Abpromise guarantee covers the use of ab5515 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 Use a concentration of 1 µg/ml. Detects a band of approximately 55 kDa (predicted molecular weight: 47 kDa).
ICC/IF Use at an assay dependent concentration.

Target

  • Function
    E3 ubiquitin ligase involved in the retrotranslocation and turnover of membrane and secretory proteins from the ER through a set of processes named ER-associated degradation (ERAD). This process acts on misfolded proteins as well as in the regulated degradation of correctly folded proteins. Enhances ionizing radiation-induced p53/TP53 stability and apoptosis via ubiquitinating MDM2 and AKT1 and decreasing AKT1 kinase activity through MDM2 and AKT1 proteasomal degradation. Regulates ER stress-induced autophagy, and may act as a tumor suppressor.
  • Pathway
    Protein modification; protein ubiquitination.
  • Sequence similarities
    Belongs to the TRIM/RBCC family.
    Contains 1 B box-type zinc finger.
    Contains 1 RING-type zinc finger.
  • Domain
    The coiled-coil domain is required for the induction of autophagy during endoplasmic reticulum (ER) stress.
    The RING-type zinc finger is required for auto-polyubiquitination.
    The C-terminal domain transmembrane domain is indispensable for the localization to the ER.
  • Post-translational
    modifications
    Auto-ubiquitinated; requires the RING-type zinc finger. Auto-polyubiquitination leads to proteasomal degradation.
  • Cellular localization
    Endoplasmic reticulum membrane. Concentrates and colocalizes with p62/SQSTM1 and ZFYVE1 at the perinuclear endoplasmic reticulum.
  • Information by UniProt
  • Database links
  • Alternative names
    • B cell chronic lymphocytic leukemia tumor suppressor Leu5 antibody
    • B-cell chronic lymphocytic leukemia tumor suppressor Leu5 antibody
    • CAR antibody
    • DLEU5 antibody
    • E3 ubiquitin-protein ligase TRIM13 antibody
    • HGNC:9976 antibody
    • LEU5 antibody
    • Leukemia associated protein 5 antibody
    • Leukemia-associated protein 5 antibody
    • Putative tumor suppressor RFP2 antibody
    • Ret finger protein 2 antibody
    • RING finger protein 77 antibody
    • RNF77 antibody
    • TRI13_HUMAN antibody
    • Trim13 antibody
    • Tripartite motif protein 13 antibody
    • Tripartite motif-containing protein 13 antibody
    see all

Images

  • Left: Endogenous RFP2
    Top: DAPI
    Bottom: ab5515

    Detection using indirect fluorescence of the signal corresponding to endogenous RFP2in 293T cells. Cells fixed using 4% formaldehyde, blocked with PBS containing 3% milk and 0.5% Triton X-100, incubated for 1 hour at 37 °C using a 1/50 dilution of antibody ab5515. Cells were then washed 3 times and incubated for 1 hour at 37 °C with a goat anti-rabbit secondary antibody (1/500 dilution) coupled to Alexa Fluor 555 (Molecular Probes). Following 3 more washes, cells were stained with DAPI and mounted with Vectashield.


    Right: Overexpressed RFP2
    Top: anti-FLAG antibody
    Middle: ab5515
    Bottom: Merge of anti-FLAG and ab5515 (and DAPI) staining.

    Detection using indirect fluorescence of the signal corresponding to staining with anti-FLAG mouse antibody (top) and antibody ab5515 against RFP2(middle) in 293T cells. 293T cells were transfected with vectors for overexpression of flagged RFP2 u

  • Detection using indirect fluorescence of the signal corresponding to endogenous RFP2 in 293T cells. Using Lipofectamine 2000 (Invitrogen), cells were transfected with either a control shRNA directed against luciferase (left, SiRluc) or a specific siRNA directed against RFP2(right). 48 hours post-transfection, cells were fixed, blocked, and incubated with a 1/50 dilution of ab5515 at 37 °C for 1 hour. Cells were then washed 3 times and incubated at 37 °C for 1 hour with a 1/500 dilution of goat anti-rabbit antibody coupled with Alexa Fluor 555 (Molecular Probes). Following 3 further washes cells were stained with DAPI and mounted with Vectashield.

    Antibody signals were extinguished when a specific RNAi was used. The few cells which retained a signal were presumably not transfected.   

  • Anti-RFP2 antibody (ab5515) at 1 µg/ml + HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 47 kDa
    Observed band size : 55 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 74 kDa. We are unsure as to the identity of these extra bands.

    Exposure time : 8 minutes

References

ab5515 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Thank you for your enquiry and I'm sorry to hear that you are experiencing difficulty with ab5515. At this point I would like to make the following suggestions. We recommend incubating with the primary for 2 hrs at RT or overnight at 4C. As you are not...

Read More

Thank you for your e-mail. I have tried to reach Dr Genevieve Fourel (Grenoble, France) to find out the siRNA sequence but unfortunately have not heard back from this researcher, I am very sorry for the inconvenience.

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

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