Anti-ZW10 antibody
| Name | Anti-ZW10 antibody |
|---|---|
| Supplier | Abcam |
| Catalog | ab21582 |
| Prices | $401.00 |
| Sizes | 100 µg |
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Applications | IHC-P WB ICC/IF ICC/IF IP |
| Species Reactivities | Human, Mouse, Rat, Dog, Xenopus |
| Antigen | Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human ZW10 |
| Blocking Peptide | ZW10 peptide |
| Description | Rabbit Polyclonal |
| Gene | ZW10 |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
All lanes : Anti-ZW10 antibody (ab21582) at 1 µg/mlLane 1 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899)Lane 2 : Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate (ab7899) with ZW10 peptide (ab23436) at 1 µg/mlLysates/proteins at 20 µg per lane.Secondary Goat polyclonal to Rabbit IgG(Alexa Fluor® 680) at 1/10,000 dilution Performed under reducing conditions.
![ZW10 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to ZW10 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab21582.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 88kDa: ZW10](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_5/25135_ZW10-Primary-antibodies-ab21582-8.jpg)
ZW10 was immunoprecipitated using 0.5mg Jurkat whole cell extract, 5µg of Rabbit polyclonal to ZW10 and 50µl of protein G magnetic beads (+). No antibody was added to the control (-). The antibody was incubated under agitation with Protein G beads for 10min, Jurkat whole cell extract lysate diluted in RIPA buffer was added to each sample and incubated for a further 10min under agitation.Proteins were eluted by addition of 40µl SDS loading buffer and incubated for 10min at 70oC; 10µl of each sample was separated on a SDS PAGE gel, transferred to a nitrocellulose membrane, blocked with 5% BSA and probed with ab21582.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 88kDa: ZW10
Panel one shows staining of HeLa cells by anti-ZW10 (ab21582) in green and DAPI in blue. The second panel shows staining by anti-ZW10 in green and SH-CREST in red, which stains the centromeres. In most cells at interphase, ZW10 is present diffusely in the cytoplasm. In prometaphase, it is associated with the kinetochore.Fix cells for 30 minutes on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with 1 mg/ml Na borohydride or 100 mM ammonium chloride in PEM. Permeablize 30 minutes with 0.5% TX-100 in PEM. Block 30 minutes in 5% milk in TBST. Primary antibody incubated overnight at 4oC diluted 1/100 in 5% milk in TBST. Secondary antibody 1 hour at RT diluted in 5% milk in TBST. Post-fix 20 min. on ice in 4% formaldehyde in PEM. Quench autofluorescence 2 x 5 minutes with ammonium chloride in PEM. Counterstain with DAPI in TBST. Mount with ProLong Gold antifade reagent from Invitrogen. Notes: Ample washing between each step. TBST = Tris buffered saline + 0.1% Tween. PE
Image courtesy of Human Protein Atlas ab215820 staining ZW10 in Human stomach, showing a strong staining pattern in gastric pits with lower levels in the basement membrane. Paraffin embedded human stomach tissue was incubated with ab21582 (1/150 dilution) for 30 mins at room temperature. Antigen retrieval was performed by heat induction in citrate buffer pH 6. ab21582 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
ab21582 (1/200) staining ZW10 in asynchronous mouse 10T1/2 cells (green). Cells were pre-extracted with PEM (1min) at room temperature, fixed in paraformaldehyde (10 mins), washed in KB+ buffer and counterstained with DAPI in order to highlight the DNA/nucleus (red). Please note that ab21582 might only perform well when using this specific protocol. Please refer to abreview for further experimental details.See Abreview
Product References
Constitutive Mad1 targeting to kinetochores uncouples checkpoint signalling from - Constitutive Mad1 targeting to kinetochores uncouples checkpoint signalling from
Maldonado M, Kapoor TM. Nat Cell Biol. 2011 Apr;13(4):475-82.
Rod/Zw10 complex is required for PIASy-dependent centromeric SUMOylation. - Rod/Zw10 complex is required for PIASy-dependent centromeric SUMOylation.
Ryu H, Azuma Y. J Biol Chem. 2010 Oct 15;285(42):32576-85.
Mitotic control of kinetochore-associated dynein and spindle orientation by human - Mitotic control of kinetochore-associated dynein and spindle orientation by human
Chan YW, Fava LL, Uldschmid A, Schmitz MH, Gerlich DW, Nigg EA, Santamaria A. J Cell Biol. 2009 Jun 1;185(5):859-74.