Anti-NAP1L1 antibody (ab21630)
Key features and details
- Rabbit polyclonal to NAP1L1
- Suitable for: IHC-P, WB, ICC/IF
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-NAP1L1 antibody
See all NAP1L1 primary antibodies -
Description
Rabbit polyclonal to NAP1L1 -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WB, ICC/IFmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Mouse, Rat, Dog, Xenopus laevis
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Immunogen
Synthetic peptide conjugated to KLH derived from within residues 350 to the C-terminus of Human NAP1L1.
Read Abcam's proprietary immunogen policy (Peptide available as ab22417.) -
Positive control
- Recombinant Human NAP1L1 protein (ab117213) can be used as a positive control in WB. 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.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As
Properties
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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
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Immunizing Peptide (Blocking)
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Isotype control
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Positive Controls
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Recombinant Protein
Applications
The Abpromise guarantee
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 (3) 1/250 - 1/1000. Detects a band of approximately 52 kDa (predicted molecular weight: 45 kDa).ICC/IF (1) Use a concentration of 1 µg/ml.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
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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
modificationsPolyglutamylated 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
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Database links
- Entrez Gene: 4673 Human
- Entrez Gene: 53605 Mouse
- Entrez Gene: 89825 Rat
- Omim: 164060 Human
- SwissProt: P55209 Human
- SwissProt: P28656 Mouse
- SwissProt: Q9Z2G8 Rat
- Unigene: 524599 Human
see all -
Alternative names
- FLJ16112 antibody
- hNRP antibody
- HSP22 like protein interacting protein antibody
see all
Images
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Western blot - Anti-NAP1L1 antibody (ab21630)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
Lanes 1 & 3 & 5 : U2OS cell lysate
Lanes 2 & 4 & 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), 49 kDa (possible cross reactivity)For each lane, U2OS cells, either treated or not treated with ionizing radiation, were scraped from a 60 mm dish and added to 75µl of 2X Laemmli buffer. 20µl of these samples were loaded into each lane. ab21630 recognizes a major band of approximately 52 kDa corresponding closely in size to NAP1L1.
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Western blot - Anti-NAP1L1 antibody (ab21630)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
All lanes : 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), 49 kDa (possible cross reactivity)
ab21630 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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NAP1L1 antibody (ab21630)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. -
Immunocytochemistry/ Immunofluorescence - Anti-NAP1L1 antibody (ab21630)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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (5)
ab21630 has been referenced in 5 publications.
- Arnold J et al. Tubulin Tyrosine Ligase Like 4 (TTLL4) overexpression in breast cancer cells is associated with brain metastasis and alters exosome biogenesis. J Exp Clin Cancer Res 39:205 (2020). PubMed: 32998758
- Wang J et al. Exosome-Mediated Transfer of lncRNA HOTTIP Promotes Cisplatin Resistance in Gastric Cancer Cells by Regulating HMGA1/miR-218 Axis. Onco Targets Ther 12:11325-11338 (2019). PubMed: 31908497
- 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). PubMed: 24526689
- Kadyrova LY et al. Human CAF-1-dependent nucleosome assembly in a defined system. Cell Cycle 12:3286-97 (2013). PubMed: 24036545
- Pourfarzad F et al. Locus-specific proteomics by TChP: targeted chromatin purification. Cell Rep 4:589-600 (2013). WB ; Human . PubMed: 23911284
Images
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Western blot - Anti-NAP1L1 antibody (ab21630)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
Lanes 1 & 3 & 5 : U2OS cell lysate
Lanes 2 & 4 & 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), 49 kDa (possible cross reactivity)For each lane, U2OS cells, either treated or not treated with ionizing radiation, were scraped from a 60 mm dish and added to 75µl of 2X Laemmli buffer. 20µl of these samples were loaded into each lane. ab21630 recognizes a major band of approximately 52 kDa corresponding closely in size to NAP1L1.
-
Western blot - Anti-NAP1L1 antibody (ab21630)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
All lanes : 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), 49 kDa (possible cross reactivity)
ab21630 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. -
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-NAP1L1 antibody (ab21630)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. -
Immunocytochemistry/ Immunofluorescence - Anti-NAP1L1 antibody (ab21630)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.
