Overview

  • Product name
  • Description
    Rabbit polyclonal to HIPPI
  • Host species
    Rabbit
  • Specificity
    This antibody has been shown to detect 0.1 ug/lane of recombinant human HIPPI. It also detects a band of the expected size when tested on a yeast lysate containing a HIPPI fusion protein.
  • Tested applications
    Suitable for: WB, ICC/IF, ChIPmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human HIPPI.

  • Positive control
    • This antibody gave a positive signal in the following tissue lysates: Human Heart Human Brain Mouse Brain 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: HeLa Nuclear

Properties

  • 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
    Preservative: 0.02% Sodium Azide
    Constituents: 1% BSA, PBS, pH 7.4
  • Concentration information loading...
  • Purity
    Immunogen affinity purified
  • Clonality
    Polyclonal
  • Isotype
    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab5205 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 1 µg/ml. Predicted molecular weight: 49 kDa.
ICC/IF Use a concentration of 5 µg/ml.
ChIP Use at an assay dependent concentration. PubMed: 21832040

Target

  • Function
    Required 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 specificity
    Present in many tissues such as brain, thymus, lymph node, lung, liver, skin and kidney (at protein level).
  • Sequence similarities
    Belongs to the IFT57 family.
  • Domain
    The pseudo DED region (pDED) meadiates the interaction with HIP1.
  • Cellular localization
    Cytoplasm > cytoskeleton > cilium basal body. Concentrates within the inner segment of cilia.
  • Information by UniProt
  • Database links
  • Alternative names
    • CDNA FLJ10147 fis clone HEMBA1003369 antibody
    • Dermal papilla derived protein 8 antibody
    • Dermal papilla-derived protein 8 antibody
    • DERP 8 antibody
    • DERP8 antibody
    • Esrrbl 1 antibody
    • Esrrbl1 antibody
    • ESRRBL1 protein antibody
    • Estrogen related receptor beta like 1 antibody
    • Estrogen related receptor beta like protein 1 antibody
    • Estrogen-related receptor beta-like protein 1 antibody
    • FLJ10147 antibody
    • Hip1 Interacting protein antibody
    • HIP1 interacting protein HIPPI antibody
    • Hip1 protein interactor antibody
    • HIP1-interacting protein antibody
    • HIPPI antibody
    • Huntingtin interacting protein 1 interacting protein antibody
    • Huntingtin interacting protein 1 protein interactor antibody
    • IFT 57 antibody
    • ift57 antibody
    • IFT57_HUMAN antibody
    • Intraflagellar transport 57 homolog (Chlamydomonas) antibody
    • Intraflagellar transport 57 homolog antibody
    • Intraflagellar transport protein 57 homolog antibody
    • MHS4R2 antibody
    • Vestrogen related receptor beta like 1 antibody
    see all

Images

  • 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

    Performed under reducing conditions.

    Predicted band size: 49 kDa
    Observed band size: 45 kDa (why is the actual band size different from the predicted?)



    Western blot using ab5205 at 1/1000 for 1hr in TBST.

    Lane 1: recombinant human HIPPI (10 ug)
    Lane 2: recombinant human HIPPI (1 ug)
    Lane 3: recombinant human HIPPI (0.1 ug)
    Lane 4: yeast extract + AD-HIPPI fusion protein
    Lane 5: yeast extract
    Lane 6: human lymphoblastoid extract (10ug)

    The indicated band in lane 4 is higher than the recombinant human HIPPI as it is a fusion protein with a Activation Domain.

     

    The indicated band in lane 4 is higher than the recombinant human HIPPI as it is a fusion protein with a Activation Domain.
  • 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).

  • 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)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 49 kDa
    Observed band size: 43,49 kDa (why is the actual band size different from the predicted?)



    This antibody recognizes a band at approximately 49 kDa in tissue lysates, however in whole cell lysates the protein appears to be running at 43 kDa. In previous studies this 43 kDa band has been blocked with the immunizing peptide (data not shown).
  • All lanes : Anti-HIPPI antibody (ab5205) at 1 µg/ml

    Lane 1 : NIH 3T3 whole cell lysate (ab7179)
    Lane 2 : MEF1 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 3 : Brain (Mouse) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 49 kDa
    Observed band size: 43,49 kDa (why is the actual band size different from the predicted?)



    This antibody recognizes a band at approximately 49 kDa in tissue lysates, however in whole cell lysates the protein appears to be running at 43 kDa.

References

This product has been referenced in:
  • Datta M & Bhattacharyya NP Regulation of RE1 Protein Silencing Transcription Factor (REST) Expression by HIP1 Protein Interactor (HIPPI). J Biol Chem 286:33759-69 (2011). ChIP, IP . 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 Res 38:878-92 (2010). WB ; Human . Read more (PubMed: 19934260) »

See all 2 Publications for this product

Customer reviews and Q&As

Application
Western blot
Loading amount
100 µg
Gel Running Conditions
Reduced Denaturing (Gradient Gel)
Sample
Human Cell lysate - whole cell (HCT116)
Specification
HCT116
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°C
Username

Abcam user community

Verified customer

Submitted Feb 18 2014

Application
Immunoprecipitation
Immuno-precipitation step
Protein A/G
Sample
Human Tissue lysate - nuclear (HCT116)
Specification
HCT116
Total protein in input
100 µg
Username

Abcam user community

Verified customer

Submitted Feb 18 2014

Abreviews
Abcam guarantees this product to work in the species/application used in this Abreview.
Application
ChIP
Sample
Human Cell lysate - nuclear (Hela Cells)
Specification
Hela Cells
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 15 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde
Detection step
Real-time PCR
Negative control
No DNA control
Username

Abcam user community

Verified customer

Submitted Jul 24 2012

Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (Hela Cells)
Total protein in input
5000 µg
Specification
Hela Cells
Immuno-precipitation step
Protein G
Username

Dr. jerry fl

Verified customer

Submitted Jul 05 2012

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Hela Cells)
Loading amount
50 µg
Specification
Hela Cells
Gel Running Conditions
Reduced Denaturing (8%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Jul 03 2012

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

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