Overview

  • Product name
  • Description
    Rabbit polyclonal to NAP1L1
  • Tested applications
    Suitable for: IP, IHC-P, WB, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human, Xenopus laevis
    Predicted to work with: Rat, Chicken, Dog
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1 - 100 of Human NAP1L1.

    (Peptide available as ab22418.)

  • Positive control
    • U2OS whole cell lysate HeLa whole cell lysate A431 whole cell lysate HEK293 whole cell lysate NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

Properties

  • Form
    Liquid
  • Storage instructions
    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • Storage buffer
    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab33076 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
IP Use at an assay dependent concentration.
IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
WB 1/250 - 1/1000. Detects a band of approximately 52 kDa (predicted molecular weight: 45 kDa).Can be blocked with Human NAP1L1 peptide (ab22418).
ICC/IF Use a concentration of 1 µg/ml.

Target

  • Function
    May be involved in modulating chromatin formation and contribute to regulation of cell proliferation.
  • Tissue specificity
    Ubiquitously expressed.
  • Sequence similarities
    Belongs to the nucleosome assembly protein (NAP) family.
  • Domain
    The acidic domains are probably involved in the interaction with histones.
  • Post-translational
    modifications
    Polyglutamylated by TTLL4, a modification that occurs exclusively on glutamate residues and results in polyglutamate chains on the gamma-carboxyl group. Some residues may also be monoglycylated but not polyglycylated due to the absence of functional TTLL10 in human.
  • Cellular localization
    Nucleus. Melanosome. Identified by mass spectrometry in melanosome fractions from stage I to stage IV.
  • Information by UniProt
  • Database links
  • Alternative names
    • FLJ16112 antibody
    • hNRP antibody
    • HSP22 like protein interacting protein antibody
    • MGC23410 antibody
    • MGC8688 antibody
    • NAP 1 antibody
    • NAP 1 related protein antibody
    • NAP 1L antibody
    • NAP-1-related protein antibody
    • NAP1 antibody
    • NAP1 L1 antibody
    • NAP1 related protein antibody
    • NAP1L antibody
    • Nap1l1 antibody
    • NAP1L1 protein antibody
    • NP1L1_HUMAN antibody
    • NRP antibody
    • Nucleosome assembly protein 1 like 1 antibody
    • Nucleosome assembly protein 1-like 1 antibody
    • Nucleosome assembly protein I-related protein antibody
    see all

Anti-NAP1L1 antibody images

  • Lanes 1 - 2 : Anti-NAP1L1 antibody (ab33076) at 1/1000 dilution
    Lanes 3 - 4 : Anti-NAP1L1 antibody (ab33076) at 1/500 dilution
    Lanes 5 - 6 : Anti-NAP1L1 antibody (ab33076) at 1/250 dilution

    Lane 1 : U2OS whole cell lysate
    Lane 2 : U2OS whole cell lysate treated with ionizing radiation
    Lane 3 : U2OS whole cell lysate
    Lane 4 : U2OS whole cell lysate treated with ionizing radiation
    Lane 5 : U2OS whole cell lysate
    Lane 6 : U2OS whole cell lysate treated with ionizing radiation


    Performed under reducing conditions.

    Predicted band size : 45 kDa
    Observed band size : 52 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 49 kDa (possible cleavage fragment,cross reactivity),64 kDa (possible cross reactivity).

    For each lane, U2OS cells, either treated or not treated with ionizing radiation, were scraped from a 60mm dish and added to 75µl of 2X Laemmli buffer. 20µl of these samples were loaded into each lane. ab33076 recognizes a major band of approximately 52 kDa corresponding closely in size to NAP1L1.

  • All lanes : Anti-NAP1L1 antibody (ab33076) at 1 µg/ml

    Lane 1 : HeLa whole cell lysate
    Lane 2 : A431 whole cell lysate
    Lane 3 : HEK293 whole cell lysate
    Lane 4 : HeLa whole cell lysate with Human NAP1L1 peptide (ab22418) at 1 µg/ml
    Lane 5 : A431 whole cell lysate with Human NAP1L1 peptide (ab22418) at 1 µg/ml
    Lane 6 : HEK293 whole cell lysate with Human NAP1L1 peptide (ab22418) at 1 µg/ml

    Lysates/proteins at 20 µg per lane.

    Secondary
    Alexa fluor goat polyclonal to rabbit IgG (700) at 1/10000 dilution
    Developed using the ECL technique

    Predicted band size : 45 kDa
    Observed band size : 52 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 49 kDa (possible cleavage fragment,cross reactivity),75 kDa (possible cross reactivity).ab33076 recognizes a major band of approximately 52 kDa corresponding closely in size to NAP1L1. This band is competed away by the addition of the immunizing peptide, suggesting that this is a specific interaction.
  • All lanes : Anti-NAP1L1 antibody (ab33076) at 1 µg/ml

    Lane 1 : NIH 3T3 whole cell lysate (ab7179)
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size : 45 kDa
    Observed band size : 52 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 74 kDa (possible cross reactivity).
  • 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.

References for Anti-NAP1L1 antibody (ab33076)

This product has been referenced in:
  • Cho I  et al. ATP-dependent chromatin remodeling by Cockayne syndrome protein B and NAP1-like histone chaperones is required for efficient transcription-coupled DNA repair. PLoS Genet 9:e1003407 (2013). IP ; Human . Read more (PubMed: 23637612) »
  • Li Z  et al. Foxa2 and H2A.Z mediate nucleosome depletion during embryonic stem cell differentiation. Cell 151:1608-16 (2012). ChIP ; Mouse . Read more (PubMed: 23260146) »

See all 4 Publications for this product

Product Wall

Thank you for your inquiry.
After blasting the immunogen sequence for ab33076, I can confirm that NAP1L4 shows no homology according to the Swiss prot blast.
Therefore it is very un-likely that the antibody cross-reacts with NAP1L4, but this ...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Xenopus laevis Cell lysate - other (egg)
Total protein in input
625 µg
Specification
egg
Immuno-precipitation step
Other - Dynabeads
Username

Abcam user community

Verified customer

Submitted Feb 26 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (Huh7-Lunet Cells)
Total protein in input
400 µg
Specification
Huh7-Lunet Cells
Immuno-precipitation step
Protein A/G
Username

Abcam user community

Verified customer

Submitted Feb 18 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Xenopus laevis Cell lysate - other (Egg)
Loading amount
25 µg
Specification
Egg
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5%
Username

Abcam user community

Verified customer

Submitted Nov 14 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (fibroblast)
Loading amount
10 µg
Specification
fibroblast
Gel Running Conditions
Non-reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Apr 13 2012

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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