Recombinant Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR13059(2)] to RYBP - ChIP Grade
- Suitable for: Flow Cyt (Intra), ChIP, IP, ICC/IF, IHC-P, WB
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-RYBP antibody [EPR13059(2)] - ChIP Grade
See all RYBP primary antibodies -
Description
Rabbit monoclonal [EPR13059(2)] to RYBP - ChIP Grade -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), ChIP, IP, ICC/IF, IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- K562, SW480, HepG2 and rat PC12 cell lysates; Human placenta and Human clear carcinoma of kidney tissues; HepG2, SW480 and K562 cells IP: NIH/3T3 cell lysates Flow Cyt (intra): A431 cells
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR13059(2) -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Isotype control
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Positive Controls
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab185971 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 |
---|---|---|
Flow Cyt (Intra) |
1/20 - 1/40.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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ChIP |
Use at an assay dependent concentration.
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IP |
1/40 - 1/60.
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ICC/IF | (1) |
1/250.
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IHC-P |
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB |
1/1000 - 1/10000. Detects a band of approximately 32 kDa (predicted molecular weight: 25 kDa).
|
Notes |
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Flow Cyt (Intra)
1/20 - 1/40. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ChIP
Use at an assay dependent concentration. |
IP
1/40 - 1/60. |
ICC/IF
1/250. |
IHC-P
1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
WB
1/1000 - 1/10000. Detects a band of approximately 32 kDa (predicted molecular weight: 25 kDa). |
Target
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Function
May be implicated in the regulation of the transcription as a repressor of the transcriptional activity of E4TF1. In tumor cell lines, may induce apoptosis. -
Tissue specificity
Widely expressed with highest levels in lymphoid tissues and placenta. -
Sequence similarities
Contains 1 RanBP2-type zinc finger. -
Cellular localization
Nucleus. Cytoplasm. Primarily found in the nucleus. - Information by UniProt
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Database links
- Entrez Gene: 23429 Human
- Entrez Gene: 56353 Mouse
- Entrez Gene: 312603 Rat
- Omim: 607535 Human
- SwissProt: Q8N488 Human
- SwissProt: Q8CCI5 Mouse
- Unigene: 7910 Human
- Unigene: 321633 Mouse
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Alternative names
- 2410018J24Rik antibody
- AAP 1 antibody
- AAP1 antibody
see all
Images
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All lanes : Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971) at 1/1000 dilution
Lane 1 : K562 cell lysate
Lane 2 : SW480 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : goat anti-rabbit IgG, (H+L), peroxidase conjugated at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 25 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted? -
Immunohistochemical analysis of paraffin-embedded, Human clear cell carcinoma kidney tissue labeling RYBP with ab185971 at a 1/100 dilution. Counter stained with hematoxylin.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunofluorescence analysis of, paraformaldehyde-fixed, HepG2 cells labeling RYBP with ab185971 at a 1/250 dilution. As secondary antibody goat anti-rabbit IgG (Alexa Fluor®555) was used at a 1/200. In blue DAPI staining.
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ab185971 (purified) at 1/20 dilution (11.7µg/mL) immunoprecipitating RYBP in NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10 µg.
Lane 1 (input): NIH/3T3 (Mouse embryonic fibroblast) whole cell lysate 10 µg
Lane 2 (+): ab185971 & NIH/3T3 whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab185971 in NIH/3T3 whole cell lysate
For western blotting, ab185971 at 1/500 dilution (0.468 µg/mL) and veriBlot for IP secondary antibody (HRP) (ab131366) at 1/1000 dilution was used.
Blocking and diluting buffer: 5% NFDM /TBST. -
Intracellular Flow Cytometry analysis of A431 (Human epidermoid carcinoma epithelial cell) cells labeling RYBP with purified ab185971 at 1/20 dilution (11.7µg/mL) (Red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) secondary antibody was used at 1/2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) (black). Unlabeled control - Unlabelled cells (blue).
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Chromatin was prepared from NIH/3T3 cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab185971 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
*http://www.abcam.com/resources?keywords=X%20ChIP%20protocol -
Intracellular Flow Cytometry analysis of paraformaldehyde-fixed K562 cells labeling RYBP with ab185971 at a 1/40 dilution and secondary antibody goat anti-rabbit IgG (FITC, red) at a 1/150 dilution, or negative control rabbit IgG (green).
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Chromatin was prepared from HEK293 cells according to the Abcam Dual X-ChIP protocol*. Cells were fixed with EGS for 30 minutes, then formaldehyde for 10 minutes.
The ChIP was performed with 25 µg of chromatin, 5 µg of ab185971 (red), and 20 µl of Protein A/G sepharose beads. 5 µg of rabbit normal IgG was added to the beads control (gray). The immunoprecipitated DNA was quantified by real time PCR (Sybr green approach).
Primers and probes are located in the first kb of the transcribed region.
*http://www.abcam.com/resources?keywords=X%20ChIP%20protocol -
Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971) at 1/10000 dilution + HepG2 cell lysate at 20 µg
Secondary
goat anti-rabbit IgG, (H+L), peroxidase conjugated at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 25 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted? -
Anti-RYBP antibody [EPR13059(2)] - ChIP Grade (ab185971) at 1/1000 dilution + rat PC12 cell lysate at 10 µg
Secondary
goat anti-rabbit IgG, (H+L), peroxidase conjugated at 1/1000 dilution
Developed using the ECL technique.
Predicted band size: 25 kDa
Observed band size: 32 kDa why is the actual band size different from the predicted? -
Immunohistochemical analysis of paraffin-embedded, Human placenta tissue labeling RYBP with ab185971 at a 1/100 dilution. Counter stained with hematoxylin.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
-
Immunofluorescence analysis of, paraformaldehyde-fixed, SW480 cells labeling RYBP with ab185971 at a 1/250 dilution. As secondary antibody goat anti-rabbit IgG (Alexa Fluor®555) was used at a 1/200. In blue DAPI staining.
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Western blot analysis on immunoprecipitation pellet from HepG2 cell lysate labeling RYBP with ab185971 at 1/50 dilution and goat anti-rabbit IgG, (H+L), peroxidase conjugated at a 1/1000 dilution.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (6)
ab185971 has been referenced in 6 publications.
- Soler M et al. The transcribed ultraconserved region uc.160+ enhances processing and A-to-I editing of the miR-376 cluster: hypermethylation improves glioma prognosis. Mol Oncol 16:648-664 (2022). PubMed: 34665919
- Sawai A et al. PRC1 sustains the integrity of neural fate in the absence of PRC2 function. Elife 11:N/A (2022). PubMed: 34994686
- Collier AJ et al. Genome-wide screening identifies Polycomb repressive complex 1.3 as an essential regulator of human naïve pluripotent cell reprogramming. Sci Adv 8:eabk0013 (2022). PubMed: 35333572
- Maybee DV et al. RYBP Sensitizes Cancer Cells to PARP Inhibitors by Regulating ATM Activity. Int J Mol Sci 23:N/A (2022). PubMed: 36233063
- Boulay G et al. The chromatin landscape of primary synovial sarcoma organoids is linked to specific epigenetic mechanisms and dependencies. Life Sci Alliance 4:N/A (2021). PubMed: 33361335
- Tian Q et al. Rybp orchestrates spermatogenesis via regulating meiosis and sperm motility in mice. Cell Cycle 19:1492-1501 (2020). PubMed: 32324084