Anti-NAP1L1 antibody
| Name | Anti-NAP1L1 antibody |
|---|---|
| Supplier | Abcam |
| Catalog | ab33076 |
| Prices | $400.00 |
| Sizes | 100 µg |
| Host | Rabbit |
| Clonality | Polyclonal |
| Isotype | IgG |
| Applications | IP IHC-P WB ICC/IF ICC/IF |
| Species Reactivities | Mouse, Human, Xenopus, Rat, Chicken, Dog |
| Antigen | Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human NAP1L1 |
| Blocking Peptide | Human NAP1L1 peptide |
| Description | Rabbit Polyclonal |
| Gene | NAP1L1 |
| Conjugate | Unconjugated |
| Supplier Page | Shop |
Product images
Lanes 1 - 2 : Anti-NAP1L1 antibody (ab33076) at 1/1000 dilutionLanes 3 - 4 : Anti-NAP1L1 antibody (ab33076) at 1/500 dilutionLanes 5 - 6 : Anti-NAP1L1 antibody (ab33076) at 1/250 dilutionLane 1 : U2OS whole cell lysateLane 2 : U2OS whole cell lysate treated with ionizing radiationLane 3 : U2OS whole cell lysateLane 4 : U2OS whole cell lysate treated with ionizing radiationLane 5 : U2OS whole cell lysateLane 6 : U2OS whole cell lysate treated with ionizing radiationPerformed under reducing conditions.
All lanes : Anti-NAP1L1 antibody (ab33076) at 1 µg/mlLane 1 : HeLa whole cell lysateLane 2 : A431 whole cell lysateLane 3 : HEK293 whole cell lysateLane 4 : HeLa whole cell lysate with Human NAP1L1 peptide (ab22418) at 1 µg/mlLane 5 : A431 whole cell lysate with Human NAP1L1 peptide (ab22418) at 1 µg/mlLane 6 : HEK293 whole cell lysate with Human NAP1L1 peptide (ab22418) at 1 µg/mlLysates/proteins at 20 µg per lane.SecondaryAlexa fluor goat polyclonal to rabbit IgG (700) at 1/10000 dilutiondeveloped using the ECL technique
All lanes : Anti-NAP1L1 antibody (ab33076) at 1 µg/mlLane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate (ab7179)Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate Lysates/proteins at 10 µg per lane.SecondaryIRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilutionPerformed under reducing conditions.
![NAP1L1 was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Rabbit polyclonal to NAP1L1 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, Hek293 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 ab33076.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 52kDa: NAP1L1; 49 kDa (possible cleavage fragment,cross reactivity.](http://www.bioprodhub.com/system/product_images/ab_products/2/sub_3/27541_NAP1L1-Primary-antibodies-ab33076-12.jpg)
NAP1L1 was immunoprecipitated using 0.5mg Hek293 whole cell extract, 5µg of Rabbit polyclonal to NAP1L1 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, Hek293 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 ab33076.Secondary: Mouse monoclonal [SB62a] Secondary Antibody to Rabbit IgG light chain (HRP) (ab99697).Band: 52kDa: NAP1L1; 49 kDa (possible cleavage fragment,cross reactivity.
IHC image of ab33076 staining in cervix formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab33076, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
ICC/IF image of ab33076 stained A431 cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab33076, 1µg/ml) overnight at +4°C. The secondary antibody (green) was ab96899, DyLight® 488 goat anti-rabbit IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.
Product References
ATP-dependent chromatin remodeling by Cockayne syndrome protein B and NAP1-like - ATP-dependent chromatin remodeling by Cockayne syndrome protein B and NAP1-like
Cho I, Tsai PF, Lake RJ, Basheer A, Fan HY. PLoS Genet. 2013 Apr;9(4):e1003407.
ATRX-mediated chromatin association of histone variant macroH2A1 regulates - ATRX-mediated chromatin association of histone variant macroH2A1 regulates
Ratnakumar K, Duarte LF, LeRoy G, Hasson D, Smeets D, Vardabasso C, Bonisch C, Zeng T, Xiang B, Zhang DY, Li H, Wang X, Hake SB, Schermelleh L, Garcia BA, Bernstein E. Genes Dev. 2012 Mar 1;26(5):433-8.
H2AZ is enriched at polycomb complex target genes in ES cells and is necessary - H2AZ is enriched at polycomb complex target genes in ES cells and is necessary
Creyghton MP, Markoulaki S, Levine SS, Hanna J, Lodato MA, Sha K, Young RA, Jaenisch R, Boyer LA. Cell. 2008 Nov 14;135(4):649-61.