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Read our guarantee »Products:Cell Biology >> Apoptosis >> Intracellular >> Survivin / IAPs
Anti-NAIP antibody
See all NAIP products (3) ...
Rabbit polyclonal to NAIP
This antibody detects recombinant human NAIP in extracts from Sf21 insect cells, but it is unknown as to whether this antibody detects the endogenous form.
Reacts with
Human
Synthetic peptide surrounding amino acid 506 of human NAIP.
Liquid
Shipped at 4°C. Upon delivery aliquot and store at -20°C or -80°C. Avoid repeated freeze / thaw cycles.
PBS pH7.2 with 50% glycerol, 1% BSA and 0.02% sodium azide
Concentration information loading...
Immunogen affinity purified
NAIP (Neuronal Apoptosis Inhibitor Protein) is a member of the IAPs (Inhibitor of Apoptosis Proteins) that function in cell death pathways to inhibit apoptosis. NAIP was discovered based on its association with a neurodegenerative disorder. Mutations and deletions in the NAIP gene locus is a contributing factor in spinal muscular atrophy. Coexpression of NAIP and hippocalcin protects neurons against calcium-induced cell death. NAIP is strongly expressed in anterior horn and motor cortex neurons of the normal brain. It is also found in human fetal neurons and in adult choroid plexus cells.
Polyclonal
IgG
Cancer >> Invasion/microenvironment >> Apoptosis >> Death receptors & ligands >> IAPs
Cell Biology >> Apoptosis >> Intracellular >> Survivin / IAPs
Western blot - NAIP antibody (ab2549)
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Our Abpromise guarantee covers the use of ab2549 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
WB: Use at a concentration of 1.0 µg/ml.
In Western blot, this antibody detects a band of approximately 140 kDa.
Not tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
Prevents motor-neuron apoptosis induced by a variety of signals. Possible role in the prevention of spinal muscular atrophy that seems to be caused by inappropriate persistence of motor-neuron apoptosis: mutated or deleted forms of NAIP have been found in individuals with severe spinal muscular atrophy.
Expressed in motor neurons, but not in sensory neurons. Found in liver and placenta, and to a lesser extent in spinal cord.
Contains 3 BIR repeats.
Contains 1 NACHT domain.
Target information above from: UniProt accessionQ13075
The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).
Western blot - NAIP antibody (ab2549)

All lanes : Anti-NAIP antibody (ab2549) at 1/1000 dilution
Lane 1 : Lysate prepared from transfected PC3 cells.
Lane 2 : Lysate prepared from non-transfected PC3 cells.
Lane 3 : Lysate prepared from transfected HeLa cells.
Lane 4 : Lysate prepared from non-transfected HeLa cells.
Lane 5 : Lysate prepared from non-transfected NTera2D1 cells.
Secondary
secondary antibody at 1/100000 dilution
Observed band size : 160 kDa (why is the actual band size different from the predicted?)
Additional bands at : 105 kDa (possible isoform),115 kDa,130 kDa (possible isoform). We are unsure as to the identity of these extra bands.
Detection of novel NAIP protein isoforms.Cells were grown in 10 cm dishes. The human PC3, NTera2D1, and HeLa cell lines were selected to screen for NAIP proteins based on preliminary RT-PCR findings. Cells transfected with the expression vector encoding the Alu-derived NAIP ORF or untransfected controls were harvested by either scraping or trypsinization following two washes with cold PBS. Cell pellets were obtained by centrifugation and resuspended in RIPA (150 mM NaCl; 1% NP-40; 0.5% sodium deoxycholate; 0.1% SDS; 50 mM Tris, pH8) and NP40 (150 mM NaCl; 1% NP-40; 50 mM Tris, pH8) lysis buffers supplemented with a protease inhibitor cocktail, and subsequently quantified using the Qubit Fluorometer. RIPA provided clearer results for the NAIP-specific antibody. Bi-phased gels containing TEMED and APS (4% stacking, 9% separating) were used to resolve total cellular protein in electrophoresis running buffer (10×: 25 mM Tris; 192 mM glycine; 0.1% SDS). Subsequently, separated proteins were transferred using a Hoefer TE 22 tank transfer unit onto Immobilon-P PVDF membrane in fresh transfer buffer (25 mM Tris, 192 mM glycine, 10% methanol, 0.1% SDS). To assess NAIP protein isoforms in primary human tissues an IMB-103-50 Instablot membrane was used. Blocking of all membranes was performed in 5% reconstituted skimmed milk powder under constant agitation at 4° overnight. The following morning, blocking solution was replaced and fresh primary antibody was applied at 1/1000 dilution for one hour at room temperature under constant agitation. Washes were carried out with TBS-T (10×: 20 mM Tris;1.4 M NaCl;1% Tween-20) at room temperature in 5 minute intervals, no more than five times. Secondary antibody was diluted in fresh TBST and 1% blocking solution to a final concentration of 1/100,000, and incubated for one hour at room temperature under constant agitation.
Image from Dr MT Romanish et al, PLoS One. 2009 Jun 2;4(6):e5761, Fig 5.
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All lanes : Anti-NAIP antibody (ab2549) at 1/1000 dilution
Lane 1 : Lysate prepared from transfected PC3 cells.
Lane 2 : Lysate prepared from non-transfected PC3 cells.
Lane 3 : Lysate prepared from transfected HeLa cells.
Lane 4 : Lysate prepared from non-transfected HeLa cells.
Lane 5 : Lysate prepared from non-transfected NTera2D1 cells.
Secondary
secondary antibody at 1/100000 dilution
Observed band size : 160 kDa (why is the actual band size different from the predicted?)
Additional bands at : 105 kDa (possible isoform),115 kDa,130 kDa (possible isoform). We are unsure as to the identity of these extra bands.
Detection of novel NAIP protein isoforms.Cells were grown in 10 cm dishes. The human PC3, NTera2D1, and HeLa cell lines were selected to screen for NAIP proteins based on preliminary RT-PCR findings. Cells transfected with the expression vector encoding the Alu-derived NAIP ORF or untransfected controls were harvested by either scraping or trypsinization following two washes with cold PBS. Cell pellets were obtained by centrifugation and resuspended in RIPA (150 mM NaCl; 1% NP-40; 0.5% sodium deoxycholate; 0.1% SDS; 50 mM Tris, pH8) and NP40 (150 mM NaCl; 1% NP-40; 50 mM Tris, pH8) lysis buffers supplemented with a protease inhibitor cocktail, and subsequently quantified using the Qubit Fluorometer. RIPA provided clearer results for the NAIP-specific antibody. Bi-phased gels containing TEMED and APS (4% stacking, 9% separating) were used to resolve total cellular protein in electrophoresis running buffer (10×: 25 mM Tris; 192 mM glycine; 0.1% SDS). Subsequently, separated proteins were transferred using a Hoefer TE 22 tank transfer unit onto Immobilon-P PVDF membrane in fresh transfer buffer (25 mM Tris, 192 mM glycine, 10% methanol, 0.1% SDS). To assess NAIP protein isoforms in primary human tissues an IMB-103-50 Instablot membrane was used. Blocking of all membranes was performed in 5% reconstituted skimmed milk powder under constant agitation at 4° overnight. The following morning, blocking solution was replaced and fresh primary antibody was applied at 1/1000 dilution for one hour at room temperature under constant agitation. Washes were carried out with TBS-T (10×: 20 mM Tris;1.4 M NaCl;1% Tween-20) at room temperature in 5 minute intervals, no more than five times. Secondary antibody was diluted in fresh TBST and 1% blocking solution to a final concentration of 1/100,000, and incubated for one hour at room temperature under constant agitation.
Image from Dr MT Romanish et al, PLoS One. 2009 Jun 2;4(6):e5761, Fig 5.
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