Loading...
You have changed your country from
to
. Please be aware that this will change the currency in the purchasing process.
Products:Epigenetics and Nuclear Signaling >> Chromatin Modifying Enzymes >> Methylation >> Lysine methylation
Overview
Product name
Anti-CLLD8/SETDB2 antibody
See all CLLD8/SETDB2 products (4) ...
Description
Rabbit polyclonal to CLLD8/SETDB2
Tested applications
WB, IHC-P, IFmore details
Cross reactivity
Reacts with
Human
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 550 - 650 of Human CLLD8/SETDB2.
(Peptide available as ab24398.)
Positive control
This antibody gave a positive signal when tested against Sf21 cells infected by baculovirus expressing CLLD8/SETDB2
General notes
Our collaborator on this project has found that the antibody works poorly in WB.
Properties
Form
Liquid
Storage instructions
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
Storage buffer
Preservative: 0.02% Sodium Azide
Constituents: 1% BSA, PBS, pH 7.4
Concentration
Concentration information loading...
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
Research areas
Epigenetics and Nuclear Signaling >> Chromatin Modifying Enzymes >> Methylation >> Lysine methylation
Applications
Show applications key
Our Abpromise guarantee covers the use of ab5517 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: Use at an assay dependent dilution. (PubMed: 20826732)
IHC-P: 1/50Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
IF
IF: 1/50IF: 1/50
Target
Function
Histone methyltransferase involved in left-right axis specification in early development and mitosis. Specifically trimethylates 'Lys-9' of histone H3 (H3K9me3). H3K9me3 is a specific tag for epigenetic transcriptional repression that recruits HP1 (CBX1, CBX3 and/or CBX5) proteins to methylated histones. Contributes to H3K9me3 in both the interspersed repetitive elements and centromere-associated repeats. Plays a role in chromosome condensation and segregation during mitosis.
Tissue specificity
Ubiquitous. Highest expression in heart, testis and ovary.
Sequence similarities
Belongs to the histone-lysine methyltransferase family.
Contains 1 MBD (methyl-CpG-binding) domain.
Contains 1 pre-SET domain.
Contains 1 SET domain.
Cellular localization
Nucleus. Chromosome.
Target information above from: UniProt accessionQ96T68
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Alternative names
- Histone-lysine N-methyltransferase SETDB2 antibody
- C13orf4 antibody
- Chromosome 13 open reading frame 4 antibody
see all
Database links
- Entrez Gene: 83852 Human
- Omim: 607865 Human
- SwissProt: Q96T68 Human
- Unigene: 631789 Human
Anti-CLLD8/SETDB2 antibody images:
Immunocytochemistry/ Immunofluorescence - CLLD8/SETDB2 antibody (ab5517)
Left: Endogenous CLLD8
Detection using indirect fluorescence of the signal corresponding to endogenous CLLD8 in 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 ab5517. 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.
Top: DAPI
Bottom: ab5517
Right: Overexpressed CLLD8
Detection using indirect fluorescence of the signal corresponding to staining with anti-FLAG mouse antibody (top) and an antibody (ab5517) against CLLD8 (middle) in 293T cells. 293T cells were transfected with vectors for overexpression of flagged CLLD8 using Lipofectamine 2000 transfection procedure (Invitrogen). Cells were fixed 48 hours post-transfection. Mouse anti-FLAG antibody was detected with a donkey anti-mouse antibody coupled with Alexa Fluor 555 (Molecular Probes) at a 1:500 dilution. ab5517 was detected using a donkey anti-rabbit antibody coupled with Alexa Fluor 488 at a 1/500 dilution.
Top: anti-FLAG antibody
Middle: ab5517
Bottom: Merge of anti-FLAG and ab5517 (and DAPI) staining.
Genevieve Fourel, Grenoble
Immunocytochemistry/ Immunofluorescence - CLLD8/SETDB2 antibody (ab5517)
Detection using indirect fluorescence of the signal corresponding to endogenous CLLD8 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 CLLD8 (right). 48 hours post-transfection, cells were fixed, blocked, and incubated with a 1:50 dilution of ab5517 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.
Genevieve Fourel, Grenoble
Western blot - CLLD8/SETDB2 antibody (ab5517)
All lanes : Anti-CLLD8/SETDB2 antibody (ab5517) at 1 µg/ml
Lane 1 : Sf21 cells infected by baculovirus containing FLAG-CLLD8 gene at 10 µg
Lane 2 : Sf21 cells infected by baculovirus containing FLAG-CLLD8 gene at 20 µg
Lane 3 : non-infected Sf21 cells at 10 µg
Lane 4 : non-infected Sf21 cells at 20 µg
Lane 5 : Sf21 infected by baculovirus containing FLAG-CLLD8 gene at 10 µg with CLLD8/SETDB2 peptide (ab24398) at 1 µg/ml
Lane 6 : Sf21 infected by baculovirus containing FLAG-CLLD8 gene at 20 µg with CLLD8/SETDB2 peptide (ab24398) at 1 µg/ml
Lane 7 : non-infected Sf21 cells at 10 µg with CLLD8/SETDB2 peptide (ab24398) at 1 µg/ml
Lane 8 : non-infected Sf21 cells at 20 µg with CLLD8/SETDB2 peptide (ab24398) at 1 µg/ml
Observed band size : 110 kDa (why is the actual band size different from the predicted?)
Additional bands at : 22 kDa (possible non-specific binding),90 kDa (possible non-specific binding).
ab5517 recognises a band corresponding to FLAG-tagged CLLD8/SETB2 at approximately 110 kDa in Sf21 cells infected by baculovirus expressing CLLD8/SETDB2 (lanes1-2). This is larger than the predicated band size (81 kDa) due to the addition of the FLAG tag.
ab5517 does not detect a band in uninfected sf21 cells (lanes3-4) or following blocking studies using the immunizing peptide (lanes5-6).
There appears to be a ladder of proteins recognised by ab5517 (lanes1-2), which could be attributed to degradation products.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - CLLD8/SETDB2 antibody (ab5517)
Image courtesy of Human Protein Atlas
ab5517 staining CLLD8 in human colon tissue and showing a distinct and strong staining of the nucleus of the glandular cells. Paraffin embedded human colon was incubated with ab5517 (1/50) for 30 minutes at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH6.
ab5517 was tested in a tissue microarray (TMA) containing a wide range of normal and cancer tissues as well as a cell microarray consisting of a range of commonly used, well characterised human cell lines. Further results for this antibody can be found at www.proteinatlas.org
References for Anti-CLLD8/SETDB2 antibody (ab5517)
This product has been referenced in:
- Hogarth CAet al. Identification and expression of potential regulators of the mammalian mitotic-to-meiotic transition. Biol Reprod 84:34-42 (2011). WB, IHC-P; Mouse.Read more (PubMed: 20826732) »
See 1 publication for this product
Publishing research using ab5517? Please let us know so that we can cite the reference in this datasheet
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"