This antibody gave a positive signal in the following tissue lysates:
This antibody gave a positive signal in the following whole cell lysates:
HeLa (Human epithelial carcinoma)
A431 (Human epithelial carcinoma)
MCF7 (Human breast adenocarcinoma)
HEK293 (Human embryonic kidney)
NIH 3T3 (Mouse embryonic fibroblast)
MEF1 (Mouse embryonic fibroblast)
This antibody gave a positive signal in the following nuclear lysates:
Storage instructionsShipped 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.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Predicted molecular weight: 49 kDa.
Use a concentration of 5 µg/ml.
Use at an assay dependent concentration. PubMed: 21832040
FunctionRequired for the formation of cilia. Plays an indirect role in sonic hedgehog signaling, cilia being required for all activity of the hedgehog pathway (By similarity). Has pro-apoptotic function via its interaction with HIP1, leading to recruit caspase-8 (CASP8) and trigger apoptosis. Has the ability to bind DNA sequence motif 5'-AAAGACATG-3' present in the promoter of caspase genes such as CASP1, CASP8 and CASP10, suggesting that it may act as a transcription regulator; however the relevance of such function remains unclear.
Tissue specificityPresent in many tissues such as brain, thymus, lymph node, lung, liver, skin and kidney (at protein level).
Sequence similaritiesBelongs to the IFT57 family.
DomainThe pseudo DED region (pDED) meadiates the interaction with HIP1.
Cellular localizationCytoplasm > cytoskeleton > cilium basal body. Concentrates within the inner segment of cilia.
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Anti-HIPPI antibody images
Western blot - Anti-HIPPI antibody (ab5205)
All lanes : Anti-HIPPI antibody (ab5205) at 1/1000 dilution
Lane 1 : Recombinant human HIPPI at 10 µg Lane 2 : Recombinant human HIPPI at 1 µg Lane 3 : Recombinant human HIPPI at 0.1 µg Lane 4 : yeast extract + AD-HIPPI fusion protein
Lane 5 : yeast extract
Lane 6 : human lymphoblastoid extract at 10 µg
ICC/IF image of ab5205 stained human HeLa cells. The cells were methanol fixed (5 min) and incubated with the antibody (ab5205, 1µg/ml) for 1h at room temperature. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Image-iTTM FX Signal Enhancer was used as the primary blocking agent, 5% BSA (in TBS-T) was used for all other blocking steps. DAPI was used to stain the cell nuclei (blue). Alexa Fluor® 594 WGA was used to label plasma membranes (red).
Western blot - HIPPI antibody (ab5205)
All lanes : Anti-HIPPI antibody (ab5205) at 1 µg/ml
Lane 1 : Human heart tissue lysate - total protein (ab29431) Lane 2 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate Lane 3 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate Lane 4 : A431 whole cell lysate (ab7909) Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate Lane 6 : HEK293 whole cell lysate (ab7902) Lane 7 : Human brain tissue lysate - total protein (ab29466)
Datta M & Bhattacharyya NP Regulation of RE1 Protein Silencing Transcription Factor (REST) Expression by HIP1 Protein Interactor (HIPPI). J Biol Chem286:33759-69 (2011).
Read more (PubMed: 21832040) »
Banerjee M et al. Transcription regulation of caspase-1 by R393 of HIPPI and its molecular partner HIP-1. Nucleic Acids Res38:878-92 (2010).
Read more (PubMed: 19934260) »