Overview

  • Product nameAnti-RFP2 antibody
    See all RFP2 primary antibodies
  • Description
    Rabbit polyclonal to RFP2
  • Tested applicationsSuitable 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

  • FunctionE3 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.
  • PathwayProtein modification; protein ubiquitination.
  • Sequence similaritiesBelongs to the TRIM/RBCC family.
    Contains 1 B box-type zinc finger.
    Contains 1 RING-type zinc finger.
  • DomainThe 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 localizationEndoplasmic 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

Anti-RFP2 antibody 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 for Anti-RFP2 antibody (ab5515)

ab5515 has not yet been referenced specifically in any publications.

Product Wall

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"