Recombinant Anti-RNF40 antibody [EPR18511] (ab191309)
![Western blot - Anti-RNF40 antibody [EPR18511] (ab191309)](https://www.abcam.com/ps/products/191/ab191309/Images/ab191309-250317-Western-blot--AntiRNF403.jpg "Western blot - Anti-RNF40 antibody [EPR18511] (ab191309)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR18511] to RNF40
- Suitable for: IHC-P, IP, WB
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-RNF40 antibody [EPR18511]
See all RNF40 primary antibodies -
Description
Rabbit monoclonal [EPR18511] to RNF40 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, IP, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
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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.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
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
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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
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR18511 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Related Products
Applications
The Abpromise guarantee
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.
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| IP |
1/80.
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| WB | (2) |
1/1000. Detects a band of approximately 130 kDa (predicted molecular weight: 114 kDa).
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| Notes |
|---|
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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. |
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WB
1/1000. Detects a band of approximately 130 kDa (predicted molecular weight: 114 kDa). |
Target
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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
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Database links
- Entrez Gene: 9810 Human
- Entrez Gene: 233900 Mouse
- Entrez Gene: 266712 Rat
- Omim: 607700 Human
- SwissProt: O75150 Human
- SwissProt: Q3U319 Mouse
- SwissProt: Q8CJB9 Rat
- Unigene: 65238 Human
see all -
Alternative names
- 95 kDa retinoblastoma associated protein antibody
- 95 kDa retinoblastoma protein binding protein antibody
- 95 kDa retinoblastoma-associated protein antibody
see all
Images
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Western blot - Anti-RNF40 antibody [EPR18511] (ab191309)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
All lanes : 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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with the MW shown by commercial antibodies from other suppliers.
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![Western blot - Anti-RNF40 antibody [EPR18511] (ab191309)](https://www.abcam.com/ps/products/191/ab191309/Images/ab191309-250316-Western-blot--AntiRNF40-2.jpg)
Western blot - Anti-RNF40 antibody [EPR18511] (ab191309)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
All lanes : 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.
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![Western blot - Anti-RNF40 antibody [EPR18511] (ab191309)](https://www.abcam.com/ps/products/191/ab191309/Images/ab191309-250315-Western-blot--AntiRNF40-1.jpg)
Western blot - Anti-RNF40 antibody [EPR18511] (ab191309)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
All lanes : 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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The molecular weight observed is consistent with the MW shown by commercial antibodies from other suppliers.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNF40 antibody [EPR18511] (ab191309)](https://www.abcam.com/ps/products/191/ab191309/Images/ab191309-250314-ihc-1.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNF40 antibody [EPR18511] (ab191309)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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNF40 antibody [EPR18511] (ab191309)](https://www.abcam.com/ps/products/191/ab191309/Images/ab191309-250313-ihc-2.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNF40 antibody [EPR18511] (ab191309)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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNF40 antibody [EPR18511] (ab191309)](https://www.abcam.com/ps/products/191/ab191309/Images/ab191309-250312-ihc-3.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-RNF40 antibody [EPR18511] (ab191309)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.
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Immunoprecipitation - Anti-RNF40 antibody [EPR18511] (ab191309)](https://www.abcam.com/ps/products/191/ab191309/Images/ab191309-250311-Immunoprecipitation--AntiRNF40.jpg)
Immunoprecipitation - Anti-RNF40 antibody [EPR18511] (ab191309)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 Detection Reagent (HRP) (ab131366), was used for detection 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. -
![Anti-RNF40 antibody [EPR18511] (ab191309)](https://www.abcam.com/ps/products/191/ab191309/Images/ab191309-8-benefits-of-recombinant-antibodies.png)
Anti-RNF40 antibody [EPR18511] (ab191309)
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (3)
ab191309 has been referenced in 3 publications.
- Oh S et al. The chromatin-binding protein PHF6 functions as an E3 ubiquitin ligase of H2BK120 via H2BK12Ac recognition for activation of trophectodermal genes. Nucleic Acids Res 48:9037-9052 (2020). PubMed: 32735658
- Wüst HM et al. Egr2-guided histone H2B monoubiquitination is required for peripheral nervous system myelination. Nucleic Acids Res 48:8959-8976 (2020). PubMed: 32672815
- Wade AK et al. LIM-domain transcription complexes interact with ring-finger ubiquitin ligases and thereby impact islet ß-cell function. J Biol Chem 294:11728-11740 (2019). PubMed: 31186351