Product datasheet
Anti-HIPPI antibody ab5205
5 Abreviews 2 References 4 Images
Overview
Product name
Anti-HIPPI antibody
Description
Rabbit polyclonal to HIPPI
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
WB, ICC/IF, ChIP
Species reactivity
Reacts with: Mouse, Human
Immunogen
Synthetic peptide conjugated to KLH derived from within residues 50 - 150 of Human
HIPPI.Read Abcam's proprietary immunogen policy
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
Purity
Immunogen affinity purified
Clonality
Polyclonal
Isotype
IgG
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
WB
Abreviews
Notes
Use a concentration of 1 µg/ml. Predicted molecular weight: 49 kDa.
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Application
Abreviews
Notes
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.
Anti-HIPPI antibody 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
Western blot - Anti-HIPPI antibody (ab5205)
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
The indicated band in lane 4 is higher than the
recombinant human HIPPI as it is a fusion
protein with a Activation Domain.
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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
Immunocytochemistry/ Immunofluorescence -
Enhancer was used as the primary blocking
HIPPI antibody (ab5205)
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 : Heart (Human) Tissue Lysate - adult
normal tissue (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 (Human) Whole Cell Lysate
(ab7909)
Western blot - HIPPI antibody (ab5205)
Lane 5 : MCF7 (Human breast
adenocarcinoma cell line) Whole Cell Lysate
Lane 6 : HEK293 (Human) Whole Cell
Lysate (ab7902)
Lane 7 : Brain (Human) Tissue Lysate - adult
normal tissue (ab29466)
Lysates/proteins at 10 µg per lane.
Secondary
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
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All lanes : Anti-HIPPI antibody (ab5205) at 1
µg/ml
Lane 1 : NIH 3T3 (Mouse) Whole Cell Lysate
(ab7179)
Lane 2 : MEF1 (Mouse embryonic fibroblast
cell line) Whole Cell Lysate
Lane 3 : Brain (Mouse) Tissue Lysate
Western blot - HIPPI antibody (ab5205)
Lysates/proteins at 10 µg per lane.
Secondary
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
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