Overview

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

    Synthetic peptide conjugated to KLH derived from within residues 350 to the C-terminus of Human NAP1L1.

    (Peptide available as ab22417.)

  • Positive control
    • This antibody gave a positive signal in the following whole cell lysates: U2OS (+/- ionizing radiation); HeLa; A431; HEK293.

      This antibody gave a positive signal in the following tissues: Formalin Fixed Paraffin Embedded Normal Human Skin.

Properties

Applications

Our Abpromise guarantee covers the use of ab21630 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
IHC-P Use a concentration of 5 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).
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 (ab21630) at 1/1000 dilution
    Lanes 3 - 4 : Anti-NAP1L1 antibody (ab21630) at 1/500 dilution
    Lanes 5 - 6 : Anti-NAP1L1 antibody (ab21630) at 1/250 dilution

    Lane 1 : U2OS cell lysate
    Lane 2 : U2OS cell lysate treated with ionizing radiation
    Lane 3 : U2OS cell lysate
    Lane 4 : U2OS cell lysate treated with ionizing radiation
    Lane 5 : U2OS cell lysate
    Lane 6 : U2OS 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).
  • All lanes : Anti-NAP1L1 antibody (ab21630) at 1 µg/ml

    Lane 1 : HeLa whole cell lysate at 20 µg
    Lane 2 : A431 whole cell lysate at 20 µg
    Lane 3 : HEK293 whole cell lysate at 20 µg
    Lane 4 : HeLa whole cell lysate at 20 µg with Human NAP1L1 peptide (ab22417) at 1 µg/ml
    Lane 5 : A431 whole cell lysate with Human NAP1L1 peptide (ab22417) at 1 µg/ml
    Lane 6 : HEK293 whole cell lysate at 20 µg with Human NAP1L1 peptide (ab22417) at 1 µg/ml

    Secondary
    Alexa fluor goat polyclonal to rabbit IgG at 1/10000 dilution

    Predicted band size : 45 kDa
    Observed band size : 52 kDa (why is the actual band size different from the predicted?)
    Additional bands at : 35 kDa (possible cross reactivity, but this band is not blocked),49 kDa (possible cleavage fragment,cross reactivity).
  • IHC image of ab21630 staining in human skin 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 ab21630, 5µg/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 ab21630 stained A431 cells. The cells were 4% formaldehyde fixed (10 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 (ab21630, 5µg/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 (ab21630)

This product has been referenced in:
  • Long L  et al. The U4/U6 recycling factor SART3 has histone chaperone activity and associates with USP15 to regulate H2B deubiquitination. J Biol Chem 289:8916-30 (2014). Read more (PubMed: 24526689) »
  • Pourfarzad F  et al. Locus-specific proteomics by TChP: targeted chromatin purification. Cell Rep 4:589-600 (2013). WB ; Human . Read more (PubMed: 23911284) »

See all 2 Publications for this product

Product Wall

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293 cells)
Gel Running Conditions
Reduced Denaturing (12,5%)
Loading amount
20 µg
Specification
HEK293 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted May 23 2017

Application
Western blot
Loading amount
10 µg
Gel Running Conditions
Reduced Denaturing (12% SDS-PAGE)
Sample
Human Cell lysate - whole cell (293T)
Specification
293T
Blocking step
Milk as blocking agent for 12 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C
Username

Abcam user community

Verified customer

Submitted Sep 25 2014

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

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