Anti-HIPPI/IFT57 antibody (ab5205)
Key features and details
- Rabbit polyclonal to HIPPI/IFT57
- Suitable for: WB, ICC/IF
- Reacts with: Mouse, Human, Recombinant fragment
- Isotype: IgG
Overview
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Product name
Anti-HIPPI/IFT57 antibody
See all HIPPI/IFT57 primary antibodies -
Description
Rabbit polyclonal to HIPPI/IFT57 -
Host species
Rabbit -
Specificity
This antibody has been shown to detect 0.1 ug/lane of recombinant human HIPPI/IFT57. It also detects a band of the expected size when tested on a yeast lysate containing a HIPPI/IFT57 fusion protein.
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Tested Applications & Species
See all applications and species dataApplication Species ICC/IF HumanWB MouseHumanRecombinant fragment
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Immunogen
Synthetic peptide corresponding to Human HIPPI/IFT57 aa 50-150 conjugated to keyhole limpet haemocyanin.
(Peptide available asab46941) -
General notes
This product was previously labelled as HIPPI
Properties
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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
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
The Abpromise guarantee
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.
GuaranteedTested applications are guaranteed to work and covered by our Abpromise guarantee.
PredictedPredicted to work for this combination of applications and species but not guaranteed.
IncompatibleDoes not work for this combination of applications and species.
Application Species ICC/IF HumanWB MouseHumanRecombinant fragmentApplication Abreviews Notes WB (2) Use a concentration of 1 µg/ml. Predicted molecular weight: 49 kDa.ICC/IF Use a concentration of 5 µg/ml.Notes WB
Use a concentration of 1 µg/ml. Predicted molecular weight: 49 kDa.ICC/IF
Use a concentration of 5 µg/ml.Target
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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
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Database links
- Entrez Gene: 55081 Human
- Entrez Gene: 73916 Mouse
- Omim: 606621 Human
- SwissProt: Q9NWB7 Human
- SwissProt: Q8BXG3 Mouse
- Unigene: 412196 Human
- Unigene: 241276 Mouse
- Unigene: 404371 Mouse
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Alternative names
- CDNA FLJ10147 fis clone HEMBA1003369 antibody
- Dermal papilla derived protein 8 antibody
- Dermal papilla-derived protein 8 antibody
see all
Images
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Western blot - Anti-HIPPI/IFT57 antibody (ab5205)All lanes : Anti-HIPPI/IFT57 antibody (ab5205) at 1/1000 dilution
Lane 1 : Recombinant human HIPPI/IFT57 at 10 µg
Lane 2 : Recombinant human HIPPI/IFT57 at 1 µg
Lane 3 : Recombinant human HIPPI/IFT57 at 0.1 µg
Lane 4 : yeast extract + AD-HIPPI/IFT57 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.
The indicated band in lane 4 is higher than the recombinant human HIPPI/IFT57 as it is a fusion protein with a Activation Domain.
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Immunocytochemistry/ Immunofluorescence - Anti-HIPPI/IFT57 antibody (ab5205)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).
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Western blot - Anti-HIPPI/IFT57 antibody (ab5205)All lanes : Anti-HIPPI/IFT57 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 :A-431 whole cell lysate (ab7909)
Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 6 :HEK-293 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). -
Western blot - Anti-HIPPI/IFT57 antibody (ab5205)All lanes : Anti-HIPPI/IFT57 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.
Protocols
Datasheets and documents
References (2)
ab5205 has been referenced in 2 publications.
- 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 . 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 . PubMed: 19934260
Images
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Western blot - Anti-HIPPI/IFT57 antibody (ab5205)All lanes : Anti-HIPPI/IFT57 antibody (ab5205) at 1/1000 dilution
Lane 1 : Recombinant human HIPPI/IFT57 at 10 µg
Lane 2 : Recombinant human HIPPI/IFT57 at 1 µg
Lane 3 : Recombinant human HIPPI/IFT57 at 0.1 µg
Lane 4 : yeast extract + AD-HIPPI/IFT57 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.
The indicated band in lane 4 is higher than the recombinant human HIPPI/IFT57 as it is a fusion protein with a Activation Domain.
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Immunocytochemistry/ Immunofluorescence - Anti-HIPPI/IFT57 antibody (ab5205)
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 - Anti-HIPPI/IFT57 antibody (ab5205)All lanes : Anti-HIPPI/IFT57 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 :A-431 whole cell lysate (ab7909)
Lane 5 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 6 :HEK-293 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). -
Western blot - Anti-HIPPI/IFT57 antibody (ab5205)All lanes : Anti-HIPPI/IFT57 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.
