Recombinant
RabMAb

Anti-RNF40 antibody [EPR18511] (ab191309)

Overview

  • Product name
    Anti-RNF40 antibody [EPR18511]
    See all RNF40 primary antibodies
  • Description
    Rabbit monoclonal [EPR18511] to RNF40
  • Tested applications
    Suitable for: IHC-P, IP, WBmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant fragment within Human RNF40 aa 200-450. The exact sequence is proprietary.
    Database link: O75150

  • Positive control
    • WB: Human fetal liver, fetal heart and fetal kidney lysates; HeLa, HEK-293, mESC, MCF7, A-375, HepG2, C6, RAW 264.7 and PC-12 whole cell lysates. IHC-P: Human, mouse and rat colon tissues. IP: HeLa whole cell lysate.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
  • Concentration information loading...
  • Purity
    Protein A purified
  • Clonality
    Monoclonal
  • Clone number
    EPR18511
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab191309 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 1/100. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
IP 1/80.
WB 1/1000. Detects a band of approximately 130 kDa (predicted molecular weight: 114 kDa).

Target

  • Function
    E3 ubiquitin-protein ligase that mediates monoubiquitination of 'Lys-120' of histone H2B (H2BK120ub1). H2BK120ub1 gives a specific tag for epigenetic transcriptional activation and is also prerequisite for histone H3 'Lys-4' and 'Lys-79' methylation (H3K4me and H3K79me, respectively). Forms a ubiquitin ligase complex in cooperation with the E2 enzyme UBE2E1/UBCH6. It thereby plays a central role in histone code and gene regulation. Required for transcriptional activation of Hox genes.
  • Tissue specificity
    Ubiquitously expressed. Expressed at higher level in testis, heart and pancreas, while it is only weakly expressed in lung, skeletal muscle and small intestine.
  • Pathway
    Protein modification; protein ubiquitination.
  • Sequence similarities
    Belongs to the BRE1 family.
    Contains 1 RING-type zinc finger.
  • Cellular localization
    Nucleus.
  • Information by UniProt
  • Database links
  • Alternative names
    • 95 kDa retinoblastoma associated protein antibody
    • 95 kDa retinoblastoma protein binding protein antibody
    • 95 kDa retinoblastoma-associated protein antibody
    • BRE 1B antibody
    • BRE1 B antibody
    • BRE1 E3 ubiquitin ligase homolog B antibody
    • BRE1-B antibody
    • BRE1B antibody
    • BRE1B_HUMAN antibody
    • DKFZp686K191 antibody
    • E3 ubiquitin protein ligase BRE1B antibody
    • E3 ubiquitin-protein ligase BRE1B antibody
    • KIAA0661 antibody
    • MGC13051 antibody
    • Rb associated protein antibody
    • RBP 95 antibody
    • RBP95 antibody
    • RING finger protein 40 antibody
    • Ring finger protein 40 E3 ubiquitin protein ligase antibody
    • RNF 40 antibody
    • Rnf40 antibody
    • STARING antibody
    see all

Images

  • All lanes : Anti-RNF40 antibody [EPR18511] (ab191309) at 1/1000 dilution

    Lane 1 : Human fetal liver lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size : 114 kDa
    Observed band size : 130 kDa (why is the actual band size different from the predicted?)


    Exposure time : 15 seconds

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The molecular weight observed is consistent with the MW shown by commercial antibodies from other suppliers.

  • All lanes : Anti-RNF40 antibody [EPR18511] (ab191309) at 1/1000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate
    Lane 2 : HEK-293 (Human epithelial cells from embryonic kidney) whole cell lysate
    Lane 3 : mESC (Mouse embryonic stem cell) whole cell lysate
    Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
    Lane 5 : A-375 (Human malignant melanoma cell line) whole cell lysate
    Lane 6 : HepG2 (Human liver hepatocellular carcinoma) whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size : 114 kDa
    Observed band size : 130 kDa (why is the actual band size different from the predicted?)

    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1, 2 and 3: 30 seconds; Lane 4, 5 and 6: 15 seconds.

    The molecular weight observed is consistent with the MW shown by commercial antibodies from other suppliers.

  • All lanes : Anti-RNF40 antibody [EPR18511] (ab191309) at 1/1000 dilution

    Lane 1 : C6 (Rat glial tumor cells) whole cell lysate
    Lane 2 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 3 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size : 114 kDa
    Observed band size : 130 kDa (why is the actual band size different from the predicted?)


    Exposure time : 15 seconds

    Blocking/Dilution buffer: 5% NFDM/TBST.

    The molecular weight observed is consistent with the MW shown by commercial antibodies from other suppliers.

  • Immunohistochemical analysis of paraffin-embedded human colon tissue labeling RNF40 with ab191309 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on epithelial cells of human colon is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling RNF40 with ab191309 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on epithelial cells of mouse colon is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

  • Immunohistochemical analysis of paraffin-embedded rat colon tissue labeling RNF40 with ab191309 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Nuclear staining on epithelial cells of rat colon is observed. Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) secondary antibody at 1/500 dilution.

  • RNF40 was immunoprecipitated from 1mg of HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysate with ab191309 at 1/80 dilution. Western blot was performed from the immunoprecipitate using ab191309 at 1/1000 dilution. VeriBlot for IP secondary antibody (HRP) (ab131366), was used as secondary antibody at 1/10000 dilution.

    Lane 1: HeLa whole cell lysate 10ug (Input). 

    Lane 2: ab191309 IP in HeLa whole cell lysate. 

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab191309 in HeLa whole cell lysate.

    Blocking and dilution buffer and concentration: 5% NFDM/TBST.
    Exposure time: 3 minutes.

References

ab191309 has not yet been referenced specifically in any publications.

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry
Sample
Human Cultured Cells (Neuroblastoma cells (SHEP, Tet21 derived))
Permeabilization
Yes - 0.1% Triton X-100
Specification
Neuroblastoma cells (SHEP, Tet21 derived)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 24°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted May 12 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (breast)
Gel Running Conditions
Reduced Denaturing (12.5)
Loading amount
100000 cells
Specification
breast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3%
Username

Abcam user community

Verified customer

Submitted Sep 26 2016

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Breast)
Gel Running Conditions
Reduced Denaturing (12.5)
Loading amount
100000 cells
Specification
Breast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3%
Username

Abcam user community

Verified customer

Submitted Sep 26 2016

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

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