HIF3A antibody (Hypoxia Inducible Factor 3, alpha Subunit) (Internal Region)

Details for Product anti-HIF3A Antibody No. ABIN250964
Antigen
  • HIF-3A
  • IPAS
  • MOP7
  • PASD7
  • bHLHe17
  • Ipas
  • NEPAS
  • Mop7
  • HIF3A
  • hypoxia inducible factor 3 alpha subunit
  • hypoxia inducible factor 3, alpha subunit
  • HIF3A
  • Hif3a
Epitope
Internal Region, AA 250-350
19
12
10
9
8
6
6
4
3
2
2
2
1
1
1
Reactivity
Human, Mouse (Murine)
82
56
34
2
Host
Rabbit
71
14
12
Clonality
Polyclonal
Conjugate
This HIF3A antibody is un-conjugated
6
5
3
2
2
2
2
2
2
2
1
1
1
1
1
1
1
1
1
Application
Immunocytochemistry (ICC), Immunofluorescence (IF), Immunoprecipitation (IP), Western Blotting (WB)
63
32
26
12
11
9
9
2
1
Options
Immunogen A synthetic peptide made to an internal region of human HIF-3 alpha (within residues 250-350). [Swiss-Prot# Q66K72]
Purification IgG purified
Plasmids, Primers & others Plasmids, Primers & others HIF3A products on genomics-online (e.g. as negative or positive controls)
Antigen
Alternative Name HIF-3 alpha (HIF3A Antibody Abstract)
Background Gene Symbol: HIF3A
Molecular Weight Theoretical MW: 68 kDa
Gene ID 64344
UniProt Q66K72
Application Notes Western Blot 1:500-1:1000, Immunocytochemistry/Immunofluorescence 1:100, Immunoprecipitation 1:10-1:500This HIF-3 alpha antibody is useful for Immunoprecipitation, Western Blot and Immunocytochemistry/Immunofluorescence. In Western Blot analysis, a band is recognized at ~68 kDa representing HIF-3 alpha. Because a band at ~71 kDa is also seen in hypoxic samples,it is important to run a negative control lysate as well (see image). In ICC/IF, cytoplasmic and nuclear staining was observed in HeLa cells, which is consistent with the literature under normoxic conditions for HIF-3 alpha (PMID: 16775626). The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.
Comment

The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.

Protocol Western Blot Protocol specific for HIF-3 alpha Antibody Western Blot Protocol
1. Perform SDS-PAGE on samples to be analyzed, loading 40 µg of total protein per lane.
. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
. Rinse the blot.
. Block the membrane using standard blocking buffer for at least 1 hour.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
. Wash the membrane in wash buffer three times for 10 minutes each.
. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
. Apply the detection reagent of choice in accordance with the manufacturers instructions.Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2 %.*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.Immunocytochemistry Protocol specific for HIF-3 alpha Antibody Immunocytochemistry ProtocolCulture cells to appropriate density in 35 mm culture dishes or 6-well plates.
. Remove culture medium and add 10 % formalin to the dish. Fix at room temperature for 30 minutes.
. Remove the formalin and add ice cold methanol. Incubate for 5-10 minutes.
. Remove methanol and add washing solution (i.e. PBS). Be sure to not let the specimen dry out. Wash three times for 10 minutes.
. To block nonspecific antibody binding incubate in 10 % normal goat serum from 1 hour to overnight at room temperature.
. Add primary antibody at appropriate dilution and incubate at room temperature from 2 hours to overnight at room temperature.
. Remove primary antibody and replace with washing solution. Wash three times for 10 minutes.
. Add secondary antibody at appropriate dilution. Incubate for 1 hour at room temperature.
. Remove antibody and replace with wash solution, then wash for 10 minutes. Add Hoechst 33258 to wash solution at 1:25,0000 and incubate for 10 minutes. Wash a third time for 10 minutes.
. Cells can be viewed directly after washing. The plates can also be stored in PBS containing Azide covered in Parafilm (TM). Cells can also be cover-slipped using Fluoromount, with appropriate sealing.*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.
Restrictions For Research Use only
Format Liquid
Buffer PBS
Buffer contains: 0.02 % Sodium Azide
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Do not freeze.
Storage 4 °C
Storage Comment Store at 4°C. Do not freeze.
Supplier Images
Western Blotting (WB) image for anti-Hypoxia Inducible Factor 3, alpha Subunit (HIF3A) (AA 250-350), (Internal Region) antibody (ABIN250964) anti-Hypoxia Inducible Factor 3, alpha Subunit (HIF3A) (AA 250-350), (Internal Region) antibody
Western Blotting (WB) image for anti-Hypoxia Inducible Factor 3, alpha Subunit (HIF3A) (AA 250-350), (Internal Region) antibody (ABIN250964) Western Blot: HIF-3 alpha Antibody [ABIN250965] - Hif3a expression in papillary renal...
Immunofluorescence (IF) image for anti-Hypoxia Inducible Factor 3, alpha Subunit (HIF3A) (AA 250-350), (Internal Region) antibody (ABIN250964) Immunocytochemistry/Immunofluorescence: HIF-3 alpha Antibody [ABIN250965] - HIF3 alph...
Product cited in: Heyboer, Milovanova, Wojcik, Grant, Chin, Hardy, Lambert, Logue, Thom: "CD34+/CD45-dim stem cell mobilization by hyperbaric oxygen - changes with oxygen dosage." in: Stem cell research, Vol. 12, Issue 3, pp. 638-45, 2014 (PubMed). (Sample species: Human). Further details: Flow Cytometry

Ando, Natsume, Iwami, Ohka, Kuchimaru, Kizaka-Kondoh, Ito, Saito, Sugita, Hoshino, Wakabayashi: "A hypoxia-inducible factor (HIF)-3α splicing variant, HIF-3α4 impairs angiogenesis in hypervascular malignant meningiomas with epigenetically silenced HIF-3α4." in: Biochemical and biophysical research communications, Vol. 433, Issue 1, pp. 139-44, 2013 (PubMed). (Sample species: Human). Further details: Western Blotting

Storti, Bolzoni, Donofrio, Airoldi, Guasco, Toscani, Martella, Lazzaretti, Mancini, Agnelli, Patrene, Maïga, Franceschi, Colla, Anderson, Neri, Amiot, Aversa, Roodman, Giuliani: "Hypoxia-inducible factor (HIF)-1α suppression in myeloma cells blocks tumoral growth in vivo inhibiting angiogenesis and bone destruction." in: Leukemia, Vol. 27, Issue 8, pp. 1697-706, 2013 (PubMed). (Sample species: Human). Further details: Western Blotting

Forooghian, Razavi, Timms: "Hypoxia-inducible factor expression in human RPE cells." in: The British journal of ophthalmology, Vol. 91, Issue 10, pp. 1406-10, 2007 (PubMed).

Wade, Guttentag, Gonzales, Maschhoff, Gonzales, Kolla, Singhal, Ballard: "Gene induction during differentiation of human pulmonary type II cells in vitro." in: American journal of respiratory cell and molecular biology, Vol. 34, Issue 6, pp. 727-37, 2006 (PubMed).

Background publications Gu, Moran, Hogenesch, Wartman, Bradfield: "Molecular characterization and chromosomal localization of a third alpha-class hypoxia inducible factor subunit, HIF3alpha." in: Gene expression, Vol. 7, Issue 3, pp. 205-13, 1999 (PubMed).

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