Overview

  • Product name
  • Description
    Rabbit polyclonal to NAIP
  • Host species
    Rabbit
  • Specificity
    ab25968 recognises Neuronal apoptosis inhibitor protein (NAIP).
  • Tested applications
    Suitable for: WB, IHC-P, ELISA, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Synthetic peptide, corresponding to 13 C terminal amino acids of Human NAIP (Genbank accession No. AAC52047).

  • Positive control
    • PC-3 cell lysate

Properties

Applications

Our Abpromise guarantee covers the use of ab25968 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
WB Use a concentration of 0.5 - 1 µg/ml. Detects a band of approximately 160 kDa (predicted molecular weight: 160 kDa).Can be blocked with Human NAIP peptide (ab39795).
IHC-P Use at an assay dependent concentration. PubMed: 20844241
ELISA Use at an assay dependent concentration.
ICC/IF Use a concentration of 10 µg/ml.

Target

  • Function
    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.
  • Tissue specificity
    Expressed in motor neurons, but not in sensory neurons. Found in liver and placenta, and to a lesser extent in spinal cord.
  • Sequence similarities
    Contains 3 BIR repeats.
    Contains 1 NACHT domain.
  • Information by UniProt
  • Database links
  • Alternative names
    • Baculoviral IAP repeat containing 1 antibody
    • Baculoviral IAP repeat-containing protein 1 antibody
    • BIRC 1 antibody
    • BIRC1 antibody
    • BIRC1_HUMAN antibody
    • Birc1a antibody
    • FLJ42520 antibody
    • NAIP antibody
    • Naip1 antibody
    • Neuronal apoptosis inhibitory protein antibody
    • NLR family apoptosis inhibitory protein antibody
    • NLR family BIR domain containing 1 antibody
    • NLRB 1 antibody
    • NLRB1 antibody
    • Nucleotide binding oligomerization domain leucine rich repeat and BIR domain containing 1 antibody
    • Psi neuronal apoptosis inhibitory protein antibody
    • psiNAIP antibody
    • Similar to occludin antibody
    see all

Images

  • Western Blot of human tonsil lysate labeling NAIP with Anti-NAIP antibody (ab25968) at 1µg/ml.
  • Western blot analysis of NAIP in PC-3 cell lysate with anti-NAIP (CT) at (A) 0.5, (B) 1, and (C) 2 µg/ml.
    Minor, lower molecular weight bands may represent alternately spliced forms.
  • ab25968 at 10µg/ml staining NAIP in A549 cells by ICC/IF

  • IHC image of ab25968 staining in human normal placenta 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 ab25968, 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.

  • Anti-NAIP antibody (ab25968) at 1/1500 dilution + Mouse RAW 264.7 cell lysate at 20 µg

    Secondary
    HRP Goat anti-rabbit IgG polyclonal at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 160 kDa
    Observed band size: 150 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 110 kDa (possible non-specific binding)


    Exposure time: 1 minute

    See Abreview

  • Immunofluorescence of NAIP in A549 cells using ab25968 at 20 ug/ml.

References

This product has been referenced in:
  • Abadía-Molina F  et al. Neuronal apoptosis inhibitory protein (NAIP) localizes to the cytokinetic machinery during cell division. Sci Rep 7:39981 (2017). Read more (PubMed: 28059125) »
  • Engel K  et al. USP9X stabilizes XIAP to regulate mitotic cell death and chemoresistance in aggressive B-cell lymphoma. EMBO Mol Med 8:851-62 (2016). Read more (PubMed: 27317434) »
See all 7 Publications for this product

Customer reviews and Q&As

1-2 of 2 Abreviews or Q&A

Application
Western blot
Sample
Mouse Cell lysate - other (RAW 264.7)
Loading amount
20 µg
Specification
RAW 264.7
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Dr. Mahesh Shivananjappa

Verified customer

Submitted Apr 27 2012

Answer

Thank you for your enquiry. I have enquired at the originator of the product for the WB protocols. Please see below for further details. I hope this information will be helpful. To answer your questions: 1. We us an anti-rabbit IgG, HRP conjugate. 2. We use a modified RIPA buffer (150 mM sodium chloride, 50 mM Tris-HCl, pH 7.4, 1 mM ethylenediaminetetraacetic acid, 1 mM phenylmethylsulfonyl fluoride, 1% Triton X-100, 1% sodium deoxycholic acid, 0.1% sodium dodecylsulfate, 5 μg/ml of aprotinin, 5 μg/ml of leupeptin). After centrifugation and determining the protein concentration, the lysate is boiled for 5 min in 1 x SDS sample buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% β-mercaptoethanol. Below are the protocols that we use. Buffer Formulations: SDS/Running Buffer: • 25 mM Tris • 192 mM Glycine • 0.1% SDS Transfer Buffer: • 20 mM Tris • 150 mM Glycine • 20% methanol 0.038% SDS Blocking Buffer • 5% non-fat dry milk • TBS Wash Buffer (TBST) • 125 mM NaCl • 25 mM Tris pH 8.0 • 0.1% Tween-20 Running Protein Samples onto a Gel 1. Cast a mini SDS-PAGE gel per your labs standard protocols or purchase pre-made gels. 2. Clamp the gel to the apparatus with per manufacturer directions. Add running buffer. 3. Remove the comb gently so as to not disturb the wells. 4. Add 6μL of your select marker to a well. 5. Add 7.5μL of lysate per well (2mg/mL for 15μg per lane). 6. Apply the anode and cathode wires to the appropriate poles and cover. 7. Run at 130V for 2 hours (or until the dye front is close to the bottom). Transfer of Proteins onto Membrane 8. Assemble the “sandwich” for the transfer apparatus per the diagram below. a. Note: Handle gel and membrane with tweezers – do not touch! 9. Pre-wet sponges and filter paper in transfer buffer. a. Note: Filter papers and membrane should be same size as the gel. 10. Insert the “sandwich” and insert into the transfer apparatus, making sure the gel is on the cathode (-) and the membrane is on the anode (+) side of the apparatus. 11. Transfer the proteins to the membrane at 250mA in transfer buffer for 2 hours 12. Remove the cassette from the apparatus, discard the gel, and place membrane on paper towels to dry. (Wash the membrane in 1xTBST to remove any leftover gel.) Blocking 13. Incubate the blot with blocking buffer overnight at 4°C or 2 hours at room temperature with gentle agitation. 14. Remove blot from blocking solution. Primary Antibody Incubation 15. Dilute antibody to the recommended dilution in 10mL of blocking buffer. 16. Incubate the blot with the primary antibody for one hour at room temperature or overnight at 4°C. 17. Wash the blot three (3) times 10 minutes each in washing buffer with gentle agitation. Secondary Antibody Incubation 18. Dilute 1μL anti-rabbit IgG-HRP conjugated secondary (or other appropriate secondary) in 10mL of blocking buffer to make a 1:10000 dilution Note: working dilution of secondary can vary from 1:2000 to 1:10000. 19. Incubate blot with secondary antibody for one (1) hour at room temperature. 20. Wash three (3) times for 10 minutes each in washing buffer with gentle agitation. Development 21. Drain wash buffer 22. Add ECL solution (Amersham) per manufacturer directions and develop for 1 minute. 23. Drain the fluid. 24. Cover the blot in plastic wrap. 25. Expose the blot to X-ray film for 1 minute in a dark room. 26. Develop the film in an X-ray processor Note: a. If there is no banding, expose the film for 5 minutes, then 30 minutes and up to overnight if the signal is weak. b. If the signal is strong, expose the film for 30 seconds or less. c. Optimal dilutions should be determined by each laboratory for each antibody

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