HIF-1 alpha Antibody (NB100-449)
HIF-1 alpha Antibody
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Distributor Data...
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anti-Hypoxia-inducible factor 1 alpha antibody, anti-Hypoxia inducible factor 1 alpha subunit basic helix antibody, Anti-Hypoxia inducible factor 1 alpha antibody, anti-HIF-1alpha antibody, anti-HIF1Alpha antibody, Anti-HIF1A antibody, anti-ARNT interacting proteinantibody, Anti-ARNT interacting protein antibody, |
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HIF-1 alpha Antibody Summary: | | Gene: | HIF1A |
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| Purity: | Immunogen affinity purified |
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| Host: | Rabbit |
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| | | Note: Not all species have been tested for usefulness with this product. Only those species listed have been tested. We cannot make any guarantees about additional reactivities which may or may not occur. |
View Additional HIF-1 alpha Products HIF-1 alpha Antibody Details: | Immunogen: | The epitope recognized by this antibody maps to a region between residues 775 and the C-terminus (residue 826) of human hypoxia-inducible factor 1 (Q16665). |
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Human, monkey, rat and mouse. Has been shown to detect HIF-1 alpha in both Mouse and Human tissue by IHC with Citrate buffer antigen retrieval. Customer reports indicate reactivity in some fish as well. |
| Uses: | This antibody is useful for: Western blot, flow cytometry and immunohistochemistry. Additionally, customers have reported success using this antibody for immunoprecipitation (see product reviews). |
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| Dilutions: | Flow Cytometry 0.125ug/10^6 cells, Immunohistochemistry-Paraffin 1:100, Immunoprecipitation 2-5 ug/mg lysate, Western Blot 1:2000 - 1:10000 |
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| Unit Size: | 0.02 mg (also available: 0.025 ml (sample size)) |
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| Concentration: | 0.2 mg/ml |
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| Positive Controls: | 2 Positive Controls |
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Packaging: | Storage: | Store at 4 °C. Do not freeze. |
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| Buffer: | Tris-buffered Saline containing 0.1% BSA. |
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| Preservative: | 0.09% Sodium Azide |
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| Limitations: | This product is for research use only and is not approved for use in humans or in clinical diagnosis. Products are guaranteed for 6 months from date of receipt, except for peptides and proteins which are guaranteed for 3 months. |
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Background: Hypoxia contributes significantly to the pathophysiology of major categories of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease.
HIF-1 is a nuclear protein involved in mammalian oxygen homeostasis. This occurs as a posttranslational modification by prolyl hydroxylation. HIF-1 is a heterodimer composed of HIF-1 alpha and HIF-1 beta subunits. Both subunits are constantly translated. However, under normoxic conditions, human HIF-1 alpha is hydroxylated at Pro402 or Pro564 by a set of HIF prolyl hydroxylases, is polyubiquinated, and eventually degraded in proteosomes. Under hypoxic conditions, the lack of hydroxylation prevents HIF degradation and increases transcriptional activity. Therefore, the concentration of HIF-1 alpha increases in the cell. In contrast, HIF-1 beta remains stable under either condition. HIF hydroxylases provide insight into hypoxic cell responses, which may be used to help isolate therapeutic targets.
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Images (4) Reviews (7) Related Products (109) Exploring HIF-1 alpha Working with HIF-1 alpha
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Jump to: Publications, Images, Reviews, Protocols/Procedures, Ask a Question
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Working with HIF-1 alpha
HIF-1 alpha Antibody Images (4) Flow Cytometry: HIF-1 alpha Antibody [NB100-449] - Hela cells were treated for 15 hrs with 200uM CoCl2, fixed in PFA, and permeabilized in 90% MeOH. 1 X 10^6 cells were stained with 0.125ug anti- HIF-alpha and secondary FITC-conjugated goat anti-rabbit (in a 150ul reaction). Black- treated, anti-KLH control IgG; Red- untreated, anti-HIF1-alpha; Blue- treated, anti-HIF1-alpha. |  Western Blot: HIF-1 alpha Antibody [NB100-449] - Detection of HIF-1 alpha in a hypoxic sample. Lane 1: CoCl2 treated Cos-7 nuclear extract (hypoxic). Lane 2: Untreated Cos-7 nuclear extract (normoxic). |  Western Blot: HIF-1 alpha antibody [NB100-449] - Detection of mouse HIF1-alpha on hypoxia treated MEFs |  Immunocytochemistry: HIF-1 alpha antibody [NB100-449] - Mouse Brain, Neurons 40X |
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Reviewed by:
Anonymous 4/8/2009
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Application:
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Western Blot
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Sample Tested:
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Mouse Neuroblastoma
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Sample Loaded:
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25ug
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Species:
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Mouse
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Lot:
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A2
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Results:
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Load:
Empty Vector Normoxia
Empty Vector Hypoxia
Hif1a over expression Normoxia
Hif1a over expression Hypoxia
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Blocking Buffer:
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5% NFDM with TBS and .5% Tween 20
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Blocking Time:
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90 min.
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Blocking Temperature:
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25 Celsius
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Dilution Ratio:
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1/500
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Incubation Time:
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Overnight-16hrs
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Incubation Temperature:
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4 Celsius
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Incubation Dilution Buffer:
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5% NFDM with TBS and .5% Tween 20
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Description:
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Santa Cruz Goat anti Rabbit Secondary
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Dilution Ratio:
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1/5000
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Method:
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Chemiluminescence onto Kodak XAR film
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Exposure Time:
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2-15 min.
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Did you observe bands?:
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Yes
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Specific Bands:
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120 kDa, 60 kDa
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Non-specific Bands:
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50 kDa
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Reviewed by:
Anonymous 3/11/2010
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Application:
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Western Blot
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Sample Tested:
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Colon Cancer Cell llines
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Sample Loaded:
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50ug
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Lot:
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A2
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Results:
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The lanes on the left are HCT116 treated with increasing concentrations of DMOG. The lanes on the right are HCT116 Hif1-/- cells with the same treatment.
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Blocking Buffer:
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Blotto
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Concentration:
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5%
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Blocking Time:
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1hr
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Blocking Temperature:
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RT
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Dilution Ratio:
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1/5000
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Incubation Time:
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overnight
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Incubation Temperature:
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4 Celsius
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Incubation Dilution Buffer:
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5% BSA in TNS with Tween
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Description:
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Cell Signaling Anti-Rabbit
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Dilution Ratio:
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1/10000
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Method:
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ECL Plus
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Did you observe bands?:
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True
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Specific Bands:
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120 kDa
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Reviewed by:
Naoharu Takano 1/3/2012
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Application:
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Western Blot
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Sample Tested:
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PC12 cell whole lysate
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Sample Loaded:
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25ug
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Species:
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Rat
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Sample Pretreated:
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hypoxia
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Lot:
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A4
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Results:
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This antibody could detect HIF-1a but weak.
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Blocking Buffer:
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5% Milk in TBST
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Concentration:
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5%
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Blocking Time:
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1hr
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Blocking Temperature:
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RT
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Dilution Ratio:
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1:1000
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Incubation Time:
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o/n
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Incubation Temperature:
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4C
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Incubation Dilution Buffer:
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blocking buffer
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Description:
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anti-Rabbit IgG
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Dilution Ratio:
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1:5000
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Method:
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ECL plus
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Did you observe bands?:
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Yes
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Reviewed by:
Cristina Rodríguez 6/8/2011
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Application:
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Western Blot
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Sample Tested:
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HUVEC whole cell lysate
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Sample Loaded:
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20 microg
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Species:
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Human
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Sample Pretreated:
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Normoxia vs Hypoxia
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Lot:
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unknown
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Results:
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Western blot analysis showing that the increase in HIF-1alpha levels triggered by hypoxia in HUVEC is prevented by a specific siRNA against HIF-1alpha (siHIFa).(N: Normoxia, H: Hypoxia; siRAND: Random siRNA
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Pub Med ID:
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21622104
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PubMedPDF:
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pub_doc_9236_672_1307536568.pdf
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Blocking Buffer:
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Milk in TBST
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Concentration:
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5%
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Blocking Time:
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overnigth
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Blocking Temperature:
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4ºC Celsius
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Dilution Ratio:
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1:500
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Incubation Time:
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1 hour
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Incubation Temperature:
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room temperature Celsius
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Incubation Dilution Buffer:
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5% Milk in TBST
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Description:
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anti-rabbit DAKO
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Manufacturer Catalog #:
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P0448
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Dilution Ratio:
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1:2000
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Method:
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SuperSignal West Dura Extended Duration Substrate
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Exposure Time:
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3 min
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Positive Control:
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Hypoxia lane in Random transfected cells
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Negative Control:
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Normoxia lane in Random transfected cells
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Did you observe bands?:
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Yes
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Specific Bands:
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Multiband pattern at aprox 110 Kda kDa
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Non-specific Bands:
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Non detected kDa
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Reviewed by:
Cemile Guldal 12/22/2010
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Application:
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Western Blot
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Sample Tested:
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cerebellar neurons
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Species:
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Mouse
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Results:
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This Hif1 alpha antibody is the only one we have found to work consistently well. We use it 1:500 in 1% BSA and reuse it for 4-5 times. We get a a doublet, the expected band at around 100KDa and another band at about 130KDa or so (right below the 150KDa marker.)
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Blocking Buffer:
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3% BSA in TBST
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Blocking Time:
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1hr
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Blocking Temperature:
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RT
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Dilution Ratio:
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1:500
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Incubation Time:
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overnight
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Incubation Temperature:
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4 Celsius
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Method:
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ECL
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Exposure Time:
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5min
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Did you observe bands?:
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Yes
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Reviewed by:
Anonymous 12/3/2010
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Application:
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IP
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Sample Tested:
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HEK 293
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Species:
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Human
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Lysate Amt:
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1mg
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Sample Pretreatment:
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100uM DFO, 24 hours
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Lot:
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A3
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Results:
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I used BD mouse anti-human HIF1 antibody for WB because I didn't want to detect the rabbit IgG from the NB100-449 antibody during the IP.
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Lysate Preclear:
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agarose A
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Diluted Lysate Concentration:
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2ug/ul
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Antibody Dilution Ratio:
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1:100
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Protein Capture:
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agarose A
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Protein Collection Method:
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4000 rpm centrifugation
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Protein Reduction:
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2X sample buffer, with SDS/bME
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Bead Removal:
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4min, 4C
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Blocking Buffer:
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5% milk in TBST
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Blocking Time:
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2 hours
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Blocking Temperature:
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4
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Dilution Ratio:
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1:2000
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Incubation Time:
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overnight
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Incubation Temperature:
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4 4
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Incubation Dilution Buffer:
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5% milk/TBST
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Description:
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anti-human IgG-HRP
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Manufacturer Catalog #:
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promega
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Dilution Ratio:
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1:10000
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Method:
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ECL
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Exposure Time:
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2 minutes
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Positive Control:
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DFO treated
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Negative Control:
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no treatment
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Did you observe bands?:
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Yes
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Specific Bands:
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116
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Additional Comments:
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I used BD mouse anti-human HIF1 antibody for IB because I didn't want to detect the rabbit IgG from the NB100-449 antibody during the IP.
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Reviewed by:
Anonymous 12/21/2009
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Application:
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Western Blot
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Sample Tested:
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human tumor model mouse
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Sample Loaded:
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12ug
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Species:
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Mouse
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Results:
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Nuclear protein extracts: Chen et al. J Cell Biochem 2001;82(3):512-521.
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Blocking Buffer:
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BSA
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Concentration:
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5%
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Blocking Time:
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60 min.
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Blocking Temperature:
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room temperature
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Dilution Ratio:
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1/500
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Incubation Time:
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overnight
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Incubation Temperature:
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4 Celsius
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Incubation Dilution Buffer:
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TBST with 5% milk
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Description:
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Anti-rabbit IgG HRP-linked, Cell Signaling #7047
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Dilution Ratio:
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1/3000
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Method:
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ECL
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Exposure Time:
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5 - 10 min.
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Did you observe bands?:
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Yes
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Specific Bands:
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100 kDa
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Publications (29)HIF-1 alpha Antibody Product Specific:- Nagao K, Oka K. HIF-2 directly activates CD82 gene expression in endothelial cells. Biochem Biophys Res Commun. 2011 Mar 4.[PMID 21382346]
- Dunford ECE, Herbst EAF, Jeoung NH, et al. Pyruvate dehydrogenase (PDH) activation during in vitro muscle contractions in PDH kinase 2 knockout mice: the effect of PDH kinase 1 compensation. Am J Physiol Regul Integr Comp Physiol. 2011 Mar 16.[PMID 21411764]
more... - Copple BL, Bai S, Burgoon LD, Moon J-O. Hypoxia-inducible factor-1[alpha] regulates the expression of genes in hypoxic hepatic stellate cells important for collagen deposition and angiogenesis. Liver Int. 2011 Feb.[PMID 20880076]
- Rabie T, Kunze R, Marti HH. Impaired hypoxic response in senescent mouse brain. Int J Dev Neurosci. 2011 Jun 16.[PMID 21704147]
- Jantsch J, Wiese M, Schödel J, et al. Toll-like receptor activation and hypoxia use distinct signaling pathways to stabilize hypoxia-inducible factor 1{alpha} (HIF1A) and result in differential HIF1A-dependent gene expression. J Leukoc Biol. 2011 Jun 17. [PMID 21685248]
- Dunford ECE, Herbst EAF, Jeoung NH, et al. Pyruvate dehydrogenase (PDH) activation during in vitro muscle contractions in PDH kinase 2 knockout mice: the effect of PDH kinase 1 compensation. Am J Physiol Regul Integr Comp Physiol. 2011 Mar 16.[ PMID: 21411764]
- Nagao K, Oka K. HIF-2 directly activates CD82 gene expression in endothelial cells.Biochem Biophys Res Commun. 2011 Apr 1;407(1):260-5. [PMID: 21382346]
- Tanaka T, Wiesener M, Bernhardt W, et al. The human HIF (hypoxia-inducible factor)-3alpha gene is a HIF-1 target gene and may modulate hypoxic gene induction.Biochem J. 2009 Oct 23;424(1):143-51. [PMID: 19694616]
- Doran DM, Kulkarni-Datar K, Cool DR et al. Hypoxia Activates Constitutive Luciferase Reporter Constructs. Biochimie. 2010 Oct 21.[ PMID: 20971156]
- Copple BL, Bai S, Moon JO. Hypoxia-inducible factor-dependent production of profibrotic mediators by hypoxic Kupffer cells.Hepatol Res. 2010 May;40(5):530-9.[ PMID: 20412331]
- Clever JL, Sakai Y, Wang RA, et al. Inefficient skeletal muscle repair in inhibitor of differentiation knockout mice suggests a crucial role for BMP signaling during adult muscle regeneration. Am J Physiol Cell Physiol. 2010 May;298(5):C1087-99.[PMID: 20181926] (Mouse, WB)
- S. K. Martin, P. Diamond, S. A. Williams, et al. Hypoxia-inducible factor-2 is a novel regulator of aberrant CXCL12 expression in multiple myeloma plasma cells. Haematologica. 2010; 95 (5): 776-784. [PMID: 20015878]
- Xiao W, Ai J, Habermacher G, et al. U19/Eaf2 Binds to Stabilizes von Hippel-Lindau Protein. Cancer Res 2009;69(6):2599-2606 [pubmed: 19258512] (WB)
- Moon J-OK, Welch TP, Gonzalez FJ, et al. Reduced liver fibrosis in hypoxia-inducible factor-1{alpha}-deficient mice. Am J Physiol Gastrointest Liver Physiol 2009;296(3):G582-592 [pubmed: 19136383] (Mouse, IHC-F/IF, WB)
- Lomb DJ, Desouza LA, Franklin JL, et al. Prolyl hydroxylase inhibitors depend on extracellular glucose HIF-2{alpha} to inhibit cell death caused by NGF deprivation: evidence that HIF-2{alpha} has a role in NGF-promoted survival of sympathetic neurons. Mol Pharmacol. 2009 May;75(5):1198-209. [pubmed: 19204094] (WB)
- Lu CW, Lin SC, Chen KF, et al. Induction of pyruvate dehydrogenase kinase-3 by hypoxia-inducible factor-1 promotes metabolic switch and drug resistance. J Biol Chem. 2008 Oct 17;283(42):28106-14. [PMID: 18718909] (Human, WB)
- Tam Kimberley, Russell Darryl, Kind Karen, Thompson Jeremy. Post Hcg Follicle Differentiation in the Mouse Is Associated with an Increase in Hypoxia Inducible Factor Activity. 41st Annual Meeting of the Society for the Study of Reproduction (SSR); May 27, 2008. Kailua Kona, HI; USA.
- Vangeison G, Carr D, Federoff HJ, et al. The good, the bad, and the cell type-specific roles of hypoxia inducible factor-1 alpha in neurons and astrocytes. J Neurosci. 2008 Feb 20;28(8):1988-93. [PMID: 18287515] (Mouse, WB)
- Becker CM, Rohwer N, Funakoshi T, et al. 2-methoxyestradiol inhibits hypoxia-inducible factor-1{alpha} and suppresses growth of lesions in a mouse model of endometriosis. Am J Pathol. 2008 Feb;172(2):534-44. [PMID: 18202195] (Mouse, IHC-P, WB)
- Sun L, Marti HH, Veltkamp R. Hyperbaric oxygen reduces tissue hypoxia and hypoxia-inducible factor-1 alpha expression in focal cerebral ischemia.Stroke. 2008 Mar;39(3):1000-6. [PMID: 18239183] (Mouse, IHC-P/IF)
- May D, Gilon D, Djonov V, et al. Transgenic system for conditional induction and rescue of chronic myocardial hibernation provides insights into genomic programs of hibernation. Proc Natl Acad Sci U S A. 2008 Jan 8;105(1):282-7. [PMID: 18162550] (Mouse, IHC-P)
- Takeda K, Aguila HL, Parikh NS, et al. Regulation of adult erythropoiesis by prolyl hydroxylase domain proteins. Blood. 2008 Mar 15;111(6):3229-35. [PMID: 18056838] (Mouse, WB)
- Glassford AJ, Yue P, Sheikh AY, et al. HIF-1 regulates hypoxia- and insulin-induced expression of apelin in adipocytes. Am J Physiol Endocrinol Metab. 2007 Dec;293(6):E1590-6. [PMID: 17878221] (Mouse, WB)
- Chen HL, Pistollato F, Hoeppner DJ, et al. Oxygen tension regulates survival and fate of mouse central nervous system precursors at multiple levels. Stem Cells. 2007 Sep;25(9):2291-301. [PMID: 17556599] (Mouse, ELISA)
- Palmer LA, Doctor A, Chhabra P, et al. S-nitrosothiols signal hypoxia-mimetic vascular pathology. J Clin Invest. 2007 Sep;117(9):2592-601. [PMID: 17786245] (Mouse, WB)
- Nagano M, Yamashita T, Hamada H, et al. Identification of functional endothelial progenitor cells suitable for the treatment of ischemic tissue using human umbilical cord blood. Blood. 2007 Jul 1;110(1):151-60. [PMID: 17379743] (Human, WB)
- Takeda K, Ho VC, Takeda H, et al. Placental but not heart defects are associated with elevated hypoxia-inducible factor alpha levels in mice lacking prolyl hydroxylase domain protein 2. Mol Cell Biol. 2006 Nov;26(22):8336-46. [PMID: 16966370] (Mouse, WB, ELISA)
- Hsu T, Adereth Y, Kose N, et al. Endocytic function of von Hippel-Lindau tumor suppressor protein regulates surface localization of fibroblast growth factor receptor 1 and cell motility. J Biol Chem. 2006 Apr 28;281(17):12069-80. [PMID: 16505488] (Human, WB)
- Litz J, Krystal GW. Imatinib inhibits c-Kit-induced hypoxia-inducible factor-1alpha activity and vascular endothelial growth factor expression in small cell lung cancer cells. Mol Cancer Ther. 2006 Jun;5(6):1415-22. [PMID: 1681849] (Human, WB)
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Jump to: Positive Controls, Antibody Packs, Antibody Pairs, Lysates, Peptides and Proteins, Primary Antibodies, RNAi, Secondary Antibodies
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Related ProductsPositive ControlsAntibody Packs (3)| Name | Catalog# | Reactivity | Applications | Images | Publications | Reviews | |
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| HIF-1 alpha Antibody Pack 5 | NB100-900WB | Hu, Mk, Mu, Rt | ChIP, FACS, ICC, IF, IHC, IHC-Fr, IHC-P, IP, WB | (11) | (36) | | | | HIF-1 alpha Antibody Pack 5Vials | NB100-905 | Hu, Mu | ChIP, FACS, ICC, IHC, IHC-Fr, IHC-P, IP, WB | (16) | (4) | | | | HIF-1 alpha Antibody Pack 4 vials | NB100-901IHC | Hu, Mu, Rt | FACS, ICC, IF, IHC, IHC-Fr, IHC-P, IP, WB | (15) | (5) | | |
Antibody Pairs (1)Lysates (1)Peptides and ProteinsPrimary Antibodies (92)| Name | Catalog# | Reactivity | Applications | Images | Publications | Reviews | |
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| HIF-1 alpha Antibody (H1alpha67) | NB100-105 | Bv, Ft, Hu, Mk, Mu, Po, Rb, Rt, Sh | ChIP, ICC, IF, IHC, IHC-Fr, IHC-P, IP, WB | (2) | (194) | | | | HIF-1 alpha Antibody | NB100-479 | Hu, Mk, Mu, Rt | ICC, IF, IHC-Fr, IHC-P, WB | (4) | (35) | | | | HIF-1 alpha Antibody (H1alpha67) | NB100-123 | Bv, Ft, Hu, Mk, Mu, Po, Rt, Sh | IF, IHC, IHC-P, IP, WB | (4) | (31) | | | | HIF-1 alpha Antibody | NB100-134 | Hu, Mk, Mu, Rt | ChIP, IHC-P, IP, WB | (3) | (31) | | | | HIF-1 alpha Antibody (ESEE122) | NB100-131 | Bv, Ca, Hu, Mu, Rt | ICC, IF, IHC-Fr, IHC-P, IP, WB | (6) | (23) | | | | ... and 87 more. See All » |
RNAi (2)Secondary Antibodies| Name | Catalog# | Reactivity | Applications | Images | Publications | Reviews | |
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| Rabbit IgG Antibody [HRP] | NB730-H | Rb | ELISA, ICC, IHC-P, WB | | (5) | | | | Rabbit IgG Antibody [FITC] | NB730-F | Rb | ICC, IHC-P | | (1) | | | | Rabbit IgG Antibody [Biotin] | NB730-B | Rb | ELISA, ICC, IHC-P, WB | | | | | | Rabbit IgG, H/L Chains Antibody [DyLight 488] | NBP1-72944 | Rb | FLISA, IF, WB | (3) | | | | | Rabbit IgG Antibody [Alkaline Phosphatase] | NB730-AP | Rb | ELISA, ICC, IHC-P, WB | | (1) | | | | See All » |
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