030_12_AH Hypoxia Card Side01.qxp_Layout 1 29/12/2014 15:15 Page 1
HIFα pathway
Co-factors:
•
•
•
•
Alpha-Ketoglutarate
Fe2+
Ascorbic acid
Oxygen
Transcriptional co-activators:
•
•
•
•
•
p300
CBP - Creb binding protein
SRC-1 - Steroid receptor co-activator 1
TIF-2 - Transcription intermediary factor 2
Ref-1 - Redox factor 1
Under normoxic conditions HIFα is hydroxylated by HIFα -specific prolyl
hydroxylases (PHD1-3), which are oxygen sensing. Hydroxylation triggers
poly-ubiquitylation of HIFα and targets it for proteasomal degradation by an
E3 ubiquitin ligase, the von Hippel-Lindau protein (pVHL) complex. Hypoxia
and TCA cycle intermediates inhibit hydroxylation and stabilize the protein.
HIFα subunits are also substrates for an asparaginyl hydroxylase
factor-inhibiting HIF1α (FIH1). This enzyme is also oxygen sensing and
hydroxylation by FIH1 disrupts a critical interaction between HIFα subunits
and co-activators such as p300/CBP, impairing HIF transcriptional activity.
Regulation of HIFα stability can also be mediated by an oxygen-independent
pathway. In the cytoplasm, HIFα is bound to heat-shock protein 90 (Hsp90)
and this association leads to enhanced stability of the subunit. However,
upon displacement of Hsp90 by Hsp90 inhibitors, receptor of activated
protein C kinase (RACK1) is able to bind and acts to recruit ubiquitin ligase
machinery and potentiate the degradation of the α-subunit.
Transcription of Target Genes:
Angiogenesis:
• Vascular endothelial growth factor
• Endothelial growth factor
Metabolism:
• Glucose transporter-1
• Hexokinase 2
• Glyceraldehyde-3 phosphate dehydrogenase
• Enolase-1
Vascular function:
• Inducible nitric oxide synthase
• Endothelial nitric oxide synthase
• Endothelin-1
Cell migration:
• C-X-C chemokine receptor type 4 (CXCR-4)
Transcription of Target Genes
Discover more at abcam.com
Produced in collaboration with Claudie Hooper, King’s College London
Erythropoiesis:
• Erythropoietin
030_12_AH Hypoxia Card Side02a:A4 30/01/2012 08:42 Page 1
Hypoxia related products from Abcam:
­
­
Clonality
Applications
Host
HIF1 alpha [EP1215Y]
HIF1 alpha (Hydroxy P402)
HIF1 alpha (Hydroxy P564)
HIF1 beta / ARNT [2B10]
HIF2 alpha
HIF3 alpha
HIF3 alpha
SIRT1
PHD1/prolyl hydroxylase [EPR2]
PHD1/prolyl hydroxylase
PHD2 / prolyl hydroxylase [EPR3661]
PHD3
SIAH1/2 [8G7H12]
HIF1AN
HIF1AN
Hsp90 [AC88]
M
P
P
M
P
P
P
M
M
M
M
M
M
M
M
M
ICC/IF, IHC-P, WB
WB
WB
ICC/IF, IHC-P, IP, WB
ICC/IF, IHC-P, WB
ICC/IF, IHC-P, WB
WB
Flow Cyt, ICC/IF, In-Cell ELISA, WB
Flow Cyt, ICC/IF, IHC-P, WB
ICC, IHC-P, IP, WB
Flow Cyt, ICC, IHC-P, IP, WB
IHC-P, WB
IP, WB
Flow Cyt, ICC/IF, IHC-P, WB
ICC/IF, WB
Flow Cyt, ICC/IF, IHC-Fr, IHC-P, IP, WB
Rb
Rb
Rb
Ms
Rb
Rb
Rb
Ms
Rb
Rb
Rb
Ms
Ms
Rb
Rb
Ms
Elongin B
TCEB1 (Elongin C)
Cullin 2
Von Hippel Lindau
ROC1
Beta-Catenin
Vascular endothelial growth factor (VEGF)
VEGF [VG-1]
EGF [EGF-2]
GLUT-1
GSK3 Beta [Y174]
Enolase-1
iNOS
eNOS
CXCR-4
P
P
P
P
P
P
P
M
M
P
M
M
P
P
P
ICC/IF, IP, WB
ICC/IF, WB
IHC-P, WB
ELISA, WB
ICC/IF, IHC-P, IP, WB
Hu, Ms, Rat, Xl
ICC/IF, IHC-FoFr, IHC-P, WB
ELISA, ICC/IF, IHC-Fr, IHC-P, IP, WB
ICC/IF, WB
ICC/IF, IHC-Fr, IHC-P, IM, IP, WB
ICC, IHC-P, WB
Flow Cyt, ICC/IF, IP, In-Cell ELISA
EM, ICC/IF, IHC-Fr, IHC-P, WB
ELISA, ICC/IF, IHC-Fr, IHC-P, IP, WB
Flow Cyt, ICC/IF, IHC-Fr, IHC-P, IP, WB
Rb
Ms
Rb
Goat
Rb
Rb
Rb
Ms
Ms
Rb
Rb
Ms
Rb
Rb
Rb
Host
Species
Hu, Rat
Hu
Hu
Hu, Ms, Rat, AGMk
Hu, Rat
Hu, Ms
Hu, Ms
Hu, Ms, Rat
Hu, Ms, Rat
Hu, Ms, Rat
Hu, Ms
Hu
Hu, Rat, Dm, Pig, Zfsh
Hu
Hu
Hu, Ms, Rat, AGMk, Ce, Carp, Chk, Dog,
Fsh, Gpig, Hm, Mk, Pig, Rb, Rtrout, Shp
Hu
Hu
Hu, Ms
Hu
Hu
ICC/IF, IHC-FoFr, IHC-P, WB, sELISA
Hu, Ms, Rat
Hu, Ms, Rat, Dog
Hu
Hu, Ms, Rat
Hu, Ms, Rat
Hu
Hu, Ms, Rat
Hu, Ms, Rat
Hu, Ms, Rat
Proteins and peptides
Applications
HIF1 alpha protein (His tag)
HIF2 alpha
PHD1/prolyl hydroxylase
Von Hippel Lindau protein (His tag)
SDS-PAGE
BL
SDS-PAGE
Panels and assay kits
Applications
Species
HIF1 alpha antibody panel
ATP synthase enzyme activity microplate assay kit
Alpha Ketoglutarate (alpha KG) assay kit
NADP/NADPH Assay Kit
IHC-Fr, IHC-P, WB
FuncS
FuncS
FuncS
HIF1 / GLUT Hypoxia ICE Kit
HIF1 / PDK1 Hypoxia ICE Kit
HIF1 / Actin Hypoxia ICE Kit
In-Cell ELISA
In-Cell ELISA
In-Cell ELISA
Hu, Ms, Rat, Fr, Pig, Rb, Sh
Hu, Rat, Cow
Sample type: Tissue culture supernatants, urine, plasma, serum
Sample type: Cell and tissue lysates, culture media, urine,
plasma and serum, as well as many other biological fluids
Hu
Hu
Hu
E. coli
E. coli
Product code
Species
Hu
ab51608
ab72775
ab72777
ab2771
ab20654
ab10134
ab2165
ab110304
ab108980
ab113077
ab109088
ab77610
ab77167
ab92498
ab12289
ab13492
ab45297
ab21778
ab2967
ab77262
ab86862
ab16051
ab46154
ab1316
ab8282
ab652
ab32391
ab112994
ab15323
ab66127
ab2074
Product code
ab82065
ab21447
ab91085
ab48702
Product code
ab110304
ab109714
ab83431
ab65349
ab125298
ab125299
ab125300
Featured antibodies
HIF1 alpha antibody [mgc3] (ab16066)
Clonality
M
Applications
EMSA, GSA, ICC/IF, IHC-P, IP, WB
Host
Ms
Species cross reactivity
Hu, Cow, Pig
ab16066 staining of HIF1 alpha in human small intestine (IBD) paraformaldehyde-fixed,
paraffin-embedded sections by immunohistochemistry.
This antibody has been cited in 7 publications and has a
rating: Excellent
HIF3 alpha antibody (ab10134)
Clonality
P
Applications
ICC/IF, IHC-P, IP, WB
Host
Rb
ICC/IF image of ab10134 stained HeLa cells (green).
This antibody has been cited in 2 publications and has a
Species cross reactivity
Hu, Ms
rating: Good
Hsp90 antibody [AC88] (ab13492)
Clonality
M
Applications
Flow Cyt, ICC/IF, IHC-Fr, IHC-P,
IP, WB
Host
Ms
Species cross reactivity
Hu, Ms, Rat, Rb, C. elegans,
Chk, Dog, Shp and more
ICC/IF image of ab13492 stained NIH3T3 mouse fibroblasts (orange).
This antibody has been cited in 2 publications and has a
rating: Good
Glucose transporter GLUT1 antibody (ab652)
Clonality
P
Applications
ICC/IF, IHC-Fr, IHC-P, IM, IP, WB
Host
Rb
ICC/IF image of ab652 stained HeLa cells (green).
This antibody has been cited in 27 publications and has a
Species cross reactivity
Hu, Ms, Rt
rating: Good
VEGF antibody (ab46154)
Clonality
P
Applications
ICC/IF, IHC-FoFr, IHC-P, WB
ICC/IF image of ab46154 stained HeLa cells (green).
This antibody has been cited in 10 publications and has a
Host
Rb
Species cross reactivity
Hu, Ms, Rt
rating: Excellent
030_12_AH
Antibodies
­
: MitoSciences product
Discover more at abcam.com
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