HIF-1 alpha Antibody

Images

 
Genetic Strategies: Knockdown Validated: HIF-1 alpha Antibody [NB100-449] - Morphological alterations induced by DMOG are HIF-1alpha dependent. glEND.2 cells were stably transfected with shGFP, shHIF-1alpha or ...read more
Biological Strategies: Western Blot: HIF-1 alpha Antibody [NB100-449] - HIF-1 alpha induction on Caki-1 cell lysate using CoCl2. Image from verified customer review.
Simple Western: HIF-1 alpha Antibody [NB100-449] - Simple Western lane view shows a specific band for HIF-1 alpha in 0.5 mg/ml of hypoxic HeLa lysate. This experiment was performed under reducing conditions using the ...read more
Immunohistochemistry: HIF-1 alpha Antibody [NB100-449] - Mouse Brain, Neurons 40X
Immunohistochemistry: HIF-1 alpha Antibody [NB100-449] - Detection of human HIF1-alpha by immunohistochemistry. Sample: FFPE section of renal cell carcinoma. Antibody: Affinity purified rabbit anti-HIF1-alpha antibody ...read more
Biological Strategies: Western Blot: HIF-1 alpha Antibody [NB100-449] - Detection of Human HIF-1 alpha by Western Blot and Immunoprecipitation. Samples: Whole cell lysate (5, 15 and 50 ug for WB; 1 mg for IP, ...read more
Western Blot: HIF-1 alpha Antibody [NB100-449] - Blot showing the effect of hypoxia on the protein expression levels of the purine biosynthetic enzymes. HIF-1 alpha is stabilized in hypoxia as expected, and no ...read more
Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody [NB100-449] - Murine primary bone marrow derived macrophages stained with HIF1-alpha antibody (red). Nuclei were counterstained with DAPI (blue). Image from ...read more
Immunohistochemistry-Paraffin: HIF-1 alpha Antibody [NB100-449] - Analysis of microvascular density and HIF-1alpha activity. Low, homogenous expression of HIF-1alpha was detected in the tumors of NNK treated mice (40X, ...read more
Flow Cytometry: HIF-1 alpha Antibody [NB100-449] - HeLa cells were treated for 15 hrs with 200 uM CoCl2, fixed in PFA, and permeabilized in 90% MeOH. 1 X 10^6 cells were stained with 0.125 ug anti-HIF-1 alpha and ...read more
Biological Strategies: Western Blot: HIF-1 alpha Antibody [NB100-449] - Analysis of HIF-1 alpha in human myeloma cell lysate using anti-HIF-1 alpha. Cells were untreated or treated with IGF-1, IL-6 or CoCl2. ...read more
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 HIF-1 alpha on hypoxia treated MEFs
Biological Strategies: Western Blot: HIF-1 alpha Antibody [NB100-449] - Samples: Whole cell lysate (5, 15 and 50 ug) from HeLa cells that were treated with cobalt chloride (+; 200 uM) or mock treated (-). ...read more
Western Blot: HIF-1 alpha Antibody [NB100-449] - BMDM were seeded at 0.5x10^6 overnight. Cells were treated with 10 ng/ml LPS for 24 hrs. Image from verified customer review.
Western Blot: HIF-1 alpha Antibody [NB100-449] - Analysis of total protein using anti-HIF-1 alpha, -CAIX, -ISCU antibodies and VDACs antibody (PMID: 29596470).
Western Blot: HIF-1 alpha Antibody [NB100-449] - Analysis of total protein using anti-HIF-1 alpha, -ISCU, -FXN antibodies. CoCl2 was used to treat or not HeLa cells for 2 days (PMID: 29596470).
Western Blot: HIF-1 alpha Antibody [NB100-449] - Detection of human HIF1 alpha by western blot. Samples: Whole cell lysate (15 and 50 ug) from HEK293T cells that were either treated with 200 uM cobalt chloride (+) or ...read more
Western Blot: HIF-1 alpha Antibody [NB100-449] - 3T3-L1 mouse embryonic fibroblast adipose-like cell line. Antibody at 1:2000. WB image submitted by a verified customer review.
Biological Strategies: Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody [NB100-449] - Formaldehyde-fixed asynchronous HeLa cells.
Immunoprecipitation: HIF-1 alpha Antibody [NB100-449] - HIF-1 alpha analysis in HEK293 cells. Image from verified customer review.
Immunoprecipitation: HIF-1 alpha Antibody [NB100-449] - Detection of human HIF1 alpha by western blot of immunoprecipitates. Samples: Whole cell lysate (1 mg for IP, 20% of IP loaded) from HEK293T cells that were ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Ca, Ch, Gt, Pm, Pm, BvSpecies Glossary
Applications WB, Simple Western, ChIP, ELISA, Flow, Flow-IC, ICC/IF, IHC, IHC-Fr, IHC-P, IP, KD, KO
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Concentration
0.2 mg/ml

Order Details

HIF-1 alpha Antibody Summary

Immunogen
The immunogen recognized by this HIF-1 alpha Antibody maps to a region between residues 775 and the C-terminus (residue 826) of human hypoxia-inducible factor 1 [Uniprot# Q16665].
Predicted Species
Bovine (100%). Backed by our 100% Guarantee.
Isotype
IgG
Clonality
Polyclonal
Host
Rabbit
Gene
HIF1A
Purity
Immunogen affinity purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Applications/Dilutions

Dilutions
  • Chromatin Immunoprecipitation 1:10 - 1:500
  • ELISA 1:100-1:2000
  • Flow (Intracellular)
  • Flow Cytometry 0.125 ug per 1 million cells in a 150 mcl reaction
  • Immunocytochemistry/Immunofluorescence 1:10-1:500
  • Immunohistochemistry 1:100 - 1:500
  • Immunohistochemistry-Frozen 1:50-1:200
  • Immunohistochemistry-Paraffin 1:50-1:200
  • Immunoprecipitation 2-5 ug/mg lysate
  • Knockdown Validated
  • Knockout Validated
  • Simple Western 1:200
  • Western Blot 1:2000 - 1:10000
Application Notes
ChIP usage was reported in scientific literature (PMID: 25557133). In Simple Western only 10 - 15 uL of the recommended dilution is used per data point. For IHC-P, Tris-EDTA pH 9.0 buffer is recommended for the heat induced epitope retrieval. ELISA (PMID: 17556599 and 16966370). Knockout data (PMID: 31793879). Use in Flow-intracellular reported in scientific literature (PMID:31722203).
Theoretical MW
93 kDa.
Disclaimer note: 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.
Control
Human Brain Cerebral Cortex Whole Tissue Lysate (Adult Whole Normal)
Human Pancreas Whole Tissue Lysate (Adult Whole Tumor)
COS-7 Nuclear Hypoxic Induced Cell Lysate
HeLa CoCl2 treated/Untreated Cell Lysate
HepG2 CoCl2 treated/Untreated Cell Lysate
HeLa Hypoxic / Normoxic Cell Lysate
HepG2 Hypoxic / Normoxic Cell Lysate
HIF-1 alpha Knockout CoCl2-treated/untreated HeLa Cell Lysate
HIF-1 alpha Knockout Hypoxic-treated/untreated HeLa Cell Lysate
NIH 3T3 CoCl2 treated/Untreated Cell Lysate
L929 CoCl2 treated/Untreated Cell Lysate
Raw 264.7 CoCl2 treated/Untreated Cell Lysate
PC12 CoCl2 treated/Untreated Cell Lysate
PC12 Hypoxic / Normoxic Cell Lysate
Raw 264.7 Hypoxic / Normoxic Cell Lysate
L929 Hypoxic / Normoxic Cell Lysate
NIH 3T3 Hypoxic / Normoxic Cell Lysate
Reviewed Applications
Read 28 Reviews rated 4.3
using
NB100-449 in the following applications:

Publications
Read Publications using
NB100-449 in the following applications:

Reactivity Notes

Monkey (COS-7) and Rat reactivities were reported in customer review feedback. Detection of HIF1 alpha in both Mouse and Human tissue by IHC. Chicken reactivity was reported in scientific literature (PMID: 25632022). Canine reactivity reported in scientific literature (PMID: 28701694). Reactivity with Goat is reported in PMID: 21599540.

Packaging, Storage & Formulations

Storage
Store at 4C. Do not freeze.
Buffer
TBS, 0.1% BSA
Preservative
0.09% Sodium Azide
Concentration
0.2 mg/ml
Purity
Immunogen affinity purified

Alternate Names for HIF-1 alpha Antibody

  • AINT
  • anti-HIF-1 alpha
  • anti-HIF1A
  • ARNT interacting protein
  • ARNT-interacting protein
  • Basic-helix-loop-helix-PAS protein MOP1
  • BHLHE78
  • Class E basic helix-loop-helix protein 78
  • HIF 1A
  • HIF1 alpha
  • HIF-1 alpha
  • HIF1
  • HIF1A
  • HIF-1a
  • HIF-1alpha
  • HIF-1-alpha
  • HIF1-alpha
  • hypoxia inducible factor 1 alpha subunit, hypoxia inducible factor 1 subunit alpha
  • hypoxia inducible factor 1, alpha subunit (basic helix-loop-helix transcription factor)
  • hypoxia-inducible factor 1-alpha
  • Member of PAS protein 1
  • member of PAS superfamily 1
  • MOP1
  • PAS domain-containing protein 8
  • PASD8

Background

Hypoxia contributes to the pathophysiology of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease (1). In cancer and particularly solid tumors, hypoxia plays a critical role in the regulation of genes involved in stem cell renewal, epithelial to mesenchymal transition (EMT), metastasis and angiogenesis. In the tumor microenvironment (TME), hypoxia influences the properties and function of stromal cells (e.g., fibroblasts, endothelial and immune cells) and is a strong determinant of tumor progression (2,3).

HIF-1 or hypoxia inducible factor 1 (predicted molecular weight 93kDa), is a transcription factor commonly referred to as a "master regulator of the hypoxic response" for its central role in the regulation of cellular adaptations to hypoxia. In its active form under hypoxic conditions, HIF-1 is stabilized by the formation of a heterodimer of HIF-1 alpha and ARNT/HIF-1 beta subunits. Nuclear HIF-1 engages p300/CBP for binding to hypoxic response elements (HREs). This process induces transcription and regulation of genes including EPO, VEGF, iNOS2, ANGPT1 and OCT4 (4,5).

Under normoxic conditions, the HIF-1 alpha subunit is rapidly targeted and degraded by the ubiquitin proteasome system. This process is mediated by prolyl hydroxylase domain enzymes (PHDs), which catalyze the hydroxylation of key proline residues (Pro-402 and Pro-564) within the oxygen-dependent degradation domain of HIF-1 alpha. Once hydroxylated, HIF-1 alpha binds the von Hippel-Lindau tumor suppressor protein (pVHL) for subsequent ubiquitination and proteasomal degradation (4). pVHL dependent regulation of HIF-1 alpha plays a role in normal physiology and disease states. Regulation of HIF-1 alpha by pVHL is critical for the suppressive function of FoxP3+ regulatory Tcells (6). Repression of pVHL expression in chronic lymphocytic leukemia (CLL) B cells leads to HIF-1 alpha stabilization and increased VEGF secretion (7).

References

1. Semenza, G. L., Agani, F., Feldser, D., Iyer, N., Kotch, L., Laughner, E., & Yu, A. (2000). Hypoxia, HIF-1, and the pathophysiology of common human diseases. Advances in Experimental Medicine and Biology.

2. Muz, B., de la Puente, P., Azab, F., & Azab, A. K. (2015). The role of hypoxia in cancer progression, angiogenesis, metastasis, and resistance to therapy. Hypoxia. https://doi.org/10.2147/hp.s93413

3. Huang, Y., Lin, D., & Taniguchi, C. M. (2017). Hypoxia inducible factor (HIF) in the tumor microenvironment: friend or foe? Science China Life Sciences. https://doi.org/10.1007/s11427-017-9178-y

4. Koyasu, S., Kobayashi, M., Goto, Y., Hiraoka, M., & Harada, H. (2018). Regulatory mechanisms of hypoxia-inducible factor 1 activity: Two decades of knowledge. Cancer Science. https://doi.org/10.1111/cas.13483

5. Dengler, V. L., Galbraith, M. D., & Espinosa, J. M. (2014). Transcriptional regulation by hypoxia inducible factors. Critical Reviews in Biochemistry and Molecular Biology. https://doi.org/10.3109/10409238.2013.838205

6. Lee, J. H., Elly, C., Park, Y., & Liu, Y. C. (2015). E3Ubiquitin Ligase VHL Regulates Hypoxia-Inducible Factor-1 alpha to Maintain Regulatory T Cell Stability and Suppressive Capacity. Immunity. https://doi.org/10.1016/j.immuni.2015.05.016

7. Ghosh, A. K., Shanafelt, T. D., Cimmino, A., Taccioli, C., Volinia, S., Liu, C. G., ... Kay, N. E. (2009). Aberrant regulation of pVHL levels by microRNA promotes the HIF/VEGF axis in CLL B cells. Blood. https://doi.org/10.1182/blood-2008-10-185686

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

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Publications for HIF-1 alpha Antibody (NB100-449)(265)

We have publications tested in 7 confirmed species: Human, Mouse, Rat, Canine, Chicken, Goat, Primate.

We have publications tested in 11 applications: ChIP, ELISA, Flow-IC, ICC/IF, IHC, IHC-Fr, IHC-P, IP, KO, PLA, WB.


Filter By Application
ChIP
(4)
ELISA
(2)
Flow-IC
(1)
ICC/IF
(10)
IHC
(21)
IHC-Fr
(7)
IHC-P
(14)
IP
(3)
KO
(1)
PLA
(1)
WB
(134)
All Applications
Filter By Species
Human
(66)
Mouse
(108)
Rat
(4)
Canine
(2)
Chicken
(4)
Goat
(1)
Primate
(1)
All Species
Showing Publications 1 - 10 of 265. Show All 265 Publications.
Publications using NB100-449 Applications Species
Marrocco A Alterations of CCSP expression and macrophages metabolism in the development of silica-induced pulmonary inflammation and fibrosis Thesis Jan 1 2020 (ICC/IF, Mouse) ICC/IF Mouse
van Dam TP DNA polymerase beta prevents AID-instigated mutagenic non-canonical mismatch DNA repair bioRxiv Jan 1 2020 (IP, Mouse) IP Mouse
Tournaire G, Loopmans S, Stegen S et al. Skeletal progenitors preserve proliferation and self-renewal upon inhibition of mitochondrial respiration by rerouting the TCA cycle Cell reports Jul 26 2022 [PMID: 35905715] (WB, Mouse) WB Mouse
Vo VTA, Kim S, Hua TNM et al. Iron commensalism of mesenchymal glioblastoma promotes ferroptosis susceptibility upon dopamine treatment Communications biology Jun 16 2022 [PMID: 35710828] (WB, Mouse) WB Mouse
Han JS, Jeon YG, Oh M et al. Adipocyte HIF2 alpha functions as a thermostat via PKA C alpha regulation in beige adipocytes Nature communications Jun 7 2022 [PMID: 35672324] (IHC, Mouse) IHC Mouse
Rashed F, Diaz-Dussan D, Mashayekhi F et al. Cellular mechanism of action of 2-nitroimidzoles as hypoxia-selective therapeutic agents Redox Biology Mar 1 2022 [PMID: 35430547] (WB, Human) WB Human
Mulangala J, Akers EJ, Solly EL et al. Pro-Calcific Environment Impairs Ischaemia-Driven Angiogenesis International journal of molecular sciences Mar 20 2022 [PMID: 35328786] (WB, Human) WB Human
Mennerich D, Kubaichuk K, Raza GS et al. ER-stress promotes VHL-independent degradation of hypoxia-inducible factors via FBXW1A/ beta TrCP Redox biology Jan 18 2022 [PMID: 35074541] (WB, Mouse, Human) WB Mouse, Human
Leinhos L Myofibroblast differentiation in hypoxia: a novel role for ArhGAP29 Thesis (WB) WB
Craps J, Joris V, Baldeschi L et al. miR-199a Downregulation as a Driver of the NOX4/HIF-1 alpha/VEGF-A Pathway in Thyroid and Orbital Adipose Tissues from Graves Patients International Journal of Molecular Sciences Dec 23 2021 [PMID: 35008579] (WB, IHC, Human) WB, IHC Human
Show All 265 Publications.

Reviews for HIF-1 alpha Antibody (NB100-449) (28) 4.328

Average Rating: 4.3
(Based on 28 reviews)
We have 28 reviews tested in 5 species: Human, Mouse, Rat, Goat, Other.

Reviews using NB100-449:
Filter by Applications
WB
(23)
IHC-P
(1)
IP
(2)
IF
(1)
Simple Western
(1)
All Applications
Filter by Species
Human
(10)
Mouse
(10)
Rat
(2)
Goat
(1)
Other
(3)
All Species
Images Ratings Applications Species Date Details
Western Blot HIF-1 alpha NB100-449
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5
reviewed by:
Raussie Baidya
WB Mouse 05/08/2020
View

Summary

ApplicationWestern Blot
Sample Tested3T3-L1 mouse embryonic fibroblast adipose-like cell line
SpeciesMouse
Lotlot A3

Comments

Commentsskimmed milk 5%
primary : 1:2000
secondary: 1: 200000
Immunohistochemistry-Paraffin HIF-1 alpha NB100-449
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4
reviewed by:
Amleto Fiocchi
IHC-P Mouse 06/07/2019
View

Summary

ApplicationImmunohistochemistry-Paraffin
Sample TestedPDX mouse
SpeciesMouse
LotLot A3

Comments

CommentsWorking Dilution 1:100, EDTA pH 9.0; detection kit Polymer-HRP.
Western Blot HIF-1 alpha NB100-449
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4
reviewed by:
Verified Customer
WB Goat 05/15/2018
View

Summary

ApplicationWestern Blot
Sample Testedlaydig cell
SpeciesGoat
LotNBP1-04676SS

Comments

Commentsdilution:1:500
Western Blot HIF-1 alpha NB100-449
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4
reviewed by:
Sarah Corcoran
WB Mouse 07/17/2017
View

Summary

ApplicationWestern Blot
Sample TestedBMDM
SpeciesMouse
LotA5
  4
reviewed by:
Ning Wang
IP Rat 04/24/2017
View

Summary

ApplicationImmunoprecipitation
Sample TestedPC-12 rat adrenal pheochromocytoma cell line
SpeciesRat
LotA6
Immunofluorescence HIF-1 alpha NB100-449
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4
reviewed by:
Anonymous -
IF Mouse 01/12/2016
View

Summary

ApplicationImmunofluorescence
Sample Testedprimary bone marrow derived macrophages
SpeciesMouse
  4
reviewed by:
Haiquan Lu
WB Human 09/07/2015
View

Summary

ApplicationWestern Blot
Sample TestedHuman cancer cell whole cell lysate
SpeciesHuman
Simple Western HIF-1 alpha NB100-449
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4
reviewed by:
Verified Customer
Simple Western Human 06/09/2015
View

Summary

ApplicationSimple Western
Sample TestedHuman mesenchymal stem cell whole cell lysate
SpeciesHuman
LotA5
Western Blot HIF-1 alpha NB100-449
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4
reviewed by:
Verified Customer
WB Human 01/09/2015
View

Summary

ApplicationWestern Blot
Sample TestedMyeloma cells whole cell lysate
SpeciesHuman
  4
reviewed by:
cynthia morimoto
WB Human 12/12/2014
View

Summary

ApplicationWestern Blot
Sample TestedSee PMID 22923663
SpeciesHuman
Western Blot HIF-1 alpha NB100-449
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4
reviewed by:
Verified Customer
WB Human 08/15/2014
View

Summary

ApplicationWestern Blot
Sample TestedHuman pancreatic cancer cell lysates
SpeciesHuman
LotA4
Western Blot HIF-1 alpha NB100-449
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5
reviewed by:
Verified Customer
WB Human 07/18/2014
View

Summary

ApplicationWestern Blot
Sample TestedCaki-1 cell lysate
SpeciesHuman
Western Blot HIF-1 alpha NB100-449
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4
reviewed by:
Lei He
WB Mouse 07/05/2014
View

Summary

ApplicationWestern Blot
Sample Testedlarge intestinal lamina propria leukocytes (LPLs).
SpeciesMouse
  5
reviewed by:
Verified Customer
WB Mouse 06/30/2014
View

Summary

ApplicationWestern Blot
Sample TestedMouse
SpeciesMouse
LotA4
Western Blot HIF-1 alpha NB100-449
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5
reviewed by:
Verified Customer
WB Human 04/29/2014
View

Summary

ApplicationWestern Blot
Sample Testednuclear fractions from T47-D beast cancer cell line
SpeciesHuman
LotA5
Western Blot HIF-1 alpha NB100-449
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4
reviewed by:
Saray Varona
WB Other 03/14/2014
View

Summary

ApplicationWestern Blot
SpeciesOther
LotA4
Western Blot HIF-1 alpha NB100-449
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4
reviewed by:
Verified Customer
WB Other 08/26/2013
View

Summary

ApplicationWestern Blot
Sample Testedpig satellite cell nuclear protein
SpeciesOther
  5
reviewed by:
Bryan Pursell
WB Human 11/05/2012
View

Summary

ApplicationWestern Blot
Sample Testedhuman cell lysates
SpeciesHuman
Western Blot HIF-1 alpha NB100-449
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5
reviewed by:
Verified Customer
WB Mouse 08/24/2012
View

Summary

ApplicationWestern Blot
Sample Testedmouse peritoneal and bone marrow derived macrophages
SpeciesMouse
LotA4
Western Blot HIF-1 alpha NB100-449
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5
reviewed by:
Verified Customer
WB 08/07/2012
View

Summary

ApplicationWestern Blot
LotA2
  4
reviewed by:
ajit shah
WB 05/09/2012
View

Summary

ApplicationWestern Blot
  4
reviewed by:
Naoharu Takano
WB Rat 01/03/2012
View

Summary

ApplicationWestern Blot
Sample TestedPC12 cell whole lysate, Sample Amount: 25ug
SpeciesRat
LotA4
Western Blot HIF-1 alpha NB100-449
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4
reviewed by:
Cristina Rodríguez
WB Human 06/08/2011
View

Summary

ApplicationWestern Blot
Sample TestedHUVEC whole cell lysate, Sample Amount: 20 microg
SpeciesHuman
Western Blot HIF-1 alpha NB100-449
Enlarge
4
reviewed by:
Cemile Guldal
WB Mouse 12/22/2010
View

Summary

ApplicationWestern Blot
Sample Testedcerebellar neurons
SpeciesMouse
Immunoprecipitation HIF-1 alpha NB100-449
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4
reviewed by:
Verified Customer
IP Human 12/03/2010
View

Summary

ApplicationImmunoprecipitation
Sample TestedHEK 293
SpeciesHuman
LotA3
  5
reviewed by:
System Admin
WB Other 03/11/2010
View

Summary

ApplicationWestern Blot
Sample TestedColon Cancer Cell lines, Sample Amount: 50ug
SpeciesOther
LotA2
  3
reviewed by:
System Admin
WB Mouse 12/21/2009
View

Summary

ApplicationWestern Blot
Sample Testedhuman tumor model mouse, Sample Amount: 12ug
SpeciesMouse
  5
reviewed by:
System Admin
WB Mouse 04/08/2009
View

Summary

ApplicationWestern Blot
Sample TestedMouse Neuroblastoma, Sample Amount: 25ug
SpeciesMouse
LotA2

Product General Protocols

Video Protocols

WB Video Protocol
ChIP Video Protocol
ChIP Webinar
ICC/IF Video Protocol

FAQs for HIF-1 alpha Antibody (NB100-449). (Showing 1 - 10 of 14 FAQs).

  1. Why is there a difference between the theoretical MW for HIF1A and the observed MW for HIF-1 alpha?
    • HIF1A, like many other proteins, has post-translational modifications. Depending on the size, amount and nature of the post-translational modifications, it can cause subtle to very large changes in molecular weight.
  2. Which antibody(ies) do you recommend for the detection of HIF-1a by immunohistochemistry in the sections of paraffin-embedded mouse liver samples? I would appreciate if you can give me several choices and rank them in the order of performance. My goal is to distinguish HIF upregulation by prolyl hydroxylase inhibitor in different liver cells.
    • All of our antibodies are of high quality and are well tested/validated in species/applications we list on the datasheet. However, we suggest the following four HIF-1 alpha antibodies based upon customer reviews, as well as the number of peer reviewed publications in which these products have been cited by researchers from reputed institutes. (1) HIF-1 alpha Antibody (H1alpha67) (cat# NB100-105) (cited in at least 218 peer reviewed publications) (2) HIF-1 alpha Antibody (cat# NB100-479) (cited in at least 51 peer reviewed publications) (3) HIF-1 alpha Antibody (H1alpha67) (cat# NB100-123 ) (cited in at least 38 peer reviewed publications) (4) HIF-1 alpha Antibody (cat# NB100-449) (cited in at least 31 peer reviewed publications).
  3. I would like to know, does a path exist for detection of HIF 1 in venous blood before and after revascularization of the leg? 
    • We are not entirely sure if HIF-1 alpha will be present in the leg after revascularization. It may be present, but you may want to search the literature to see if this has been looked at before. If not, then this would certainly be an experiment worth doing.
  4. What is the molecular weight (kDa) of protein HIF 1 alpha in western blot?
    • The theoretical molecular weight of HIF 1-alpha is ~93kDa. However, you will likely see a band between 100-120kDa due to phosphorylation.
  5. We got the Hif1a (https://www.novusbio.com/products/hif-1-alpha-antibody-h1alpha67_nb100-105 ) antibody from you guys. I used the concentration that is mentioned on your website, but I am getting a band of a completely different size (~70kDa) and not the 120 kDa mentioned.
    • HIF-1 alpha is a notoriously difficult protein to work with due to its rapid degradation. Therefore, the ~70kDa bands are most likely degradation products. It is very important to lyse the cells in hypoxic conditions. We strongly recommend lysing the cells directly into the Laemmli buffer and doing that quickly, so that the exposure to oxygen is minimized.Please go through our hypoxia related FAQs, you should find them very informative:https://www.novusbio.com/support/hypoxia-and-hif-faqsAlso, running a positive control may help confirm the band specificity in your samples. You may prepare them yourself or choose some from our catalog, for example: 1) HeLa Hypoxic / Normoxic Cell Lysate (NBP2-36452)2) HeLa Hypoxic (CoCl2) / Normoxic Cell Lysate (NBP2-36450)
  6. I performed several Western Blots of HIF-1 alpha with different lysis buffers, whole lysates, and cytoplasm/nuclei extractions. I can’t seem to get a good western blot (poor signal, band much lower than expected, etc.). Can someone suggest some technical considerations/tricks I should consider using?
    • A major issue that researchers working with HIF-1 alpha is degradation due to exposure to oxygen. In western blot, this results in a weaker band and/or the appearance of multiple low molecular weight bands (40-80 kDa). We recommend preparing the lysates after collection of cells/tissues as quickly as possible (on ice), preferably in a hypoxic chamber. We also recommend including a true hypoxia mimetic control (eg: cells treated with CoCl2, DMOG… etc.). The controls help distinguish your band of interest from potential degradation/dimer bands.For more troubleshooting tips and frequently asked questions regarding hypoxia/HIFs, you can refer to our hypoxia-related FAQs: https://www.novusbio.com/support/hypoxia-and-hif-faqs
  7. I am doing HIF1 westerns in HIF-overexpressing mouse liver and adipose tissue using Novus antirabbit HIF1a antibody with overnight incubation. I am getting strong bands around 90kDa. I am aware that HIF theoretical molecular weight is 93kDa, but in westerns, the HIF band is usually around 120kDa according to my internet research. Can someone let me know if I’m getting the right HIF band or just some non-specific bands? Thanks.
    • (1)    HIF-1 alpha’s theoretical molecular weight is 93kDa. The post translationally modified/ubiquitinated form of HIF-1 alpha protein (fails to undergo proteasomal degradation) shows up as a band in the 110-130 kDa range on a Western blot.(2)    The dimeric protein may appear at a position above 200 kDa on non-reducing gels.(3)    Importantly, HIFs are among the most rapidly degradable proteins; therefore, sample preparation is highly important when analyzing HIF1 alpha or HIF2 alpha. When degraded, HIF-1 alpha may show up between 40-80 kDa position on Western blot. Degradation may be avoided by preparing the samples as soon as possible after collection of cells/tissues in hypoxic chamber. Notably, the tissues/cells should be kept on ice during lysate preparation and the lysates should be analyzed as soon as possible.(4)    For troubleshooting suggestions/feedback on more than 25 similar frequently asked questions, I would recommend visiting Novus page: FAQs - Hypoxia and HIFs https://www.novusbio.com/support/hypoxia-and-hif-faqs(5)    Last but not the least, Novus technical support team may be contacted at: technical@novusbio.com 
  8. I have Hif1a nuclear protein extract at -80C. I am wondering if anyone knows how long it would be good for at that temperature since HIf1a is known to be degraded easily.Thank you!
    • You could try a few things to further inhibit the degradation.1) Use the protease inhibitors (if you are not already using them).2) Lyse cells into a buffer that contains SDS or LDS (eg: Laemmli's buffer), since SDS and LDS denature and inhibit proteases. Lysis may even be performed with reducing agents in the buffer (eg. DTT), but this will make your lysates unsuitable for BCA assay.3) Lysing samples rapidly ensures that the samples are instantly homogenized (it also shears DNA released by the SDS).5) Flash-freezing samples in liquid nitrogen rather than freezing at -80*C reduces the window of time for protease activity.6) Freeze samples in individual aliquots, instead of thawing the same vial multiple times.
  9. I am curious to know the biochemical reactions of CoCl2 that mimic hypoxia. Is it that CoCl2 can bind any ubiquitin enzyme which regulates their degradation?
    • CoCl2 inhibits PHD enzymes (the body’s “oxygen sensors”) by replacing the Fe ion with Co, preventing these enzymes from marking HIF-1 alpha for degradation. CoCl2-based hypoxia mimetic samples are often used as positive control in HIF analysis. For more troubleshooting tips and frequently asked questions regarding hypoxia/HIFs, you can refer to our hypoxia-related FAQs: http://www.novusbio.com/support/hypoxia-and-hif-faqs
  10. I am curious to know the biochemical reactions of CoCl2 that mimic hypoxia. Is it that CoCl2 can bind any ubiquitin enzyme which regulates their degradation?
    • CoCl2 inhibits PHD enzymes (the body’s “oxygen sensors”) by replacing the Fe ion with Co, preventing these enzymes from marking HIF-1 alpha for degradation. CoCl2-based hypoxia mimetic samples are often used as positive control in HIF analysis. For more troubleshooting tips and frequently asked questions regarding hypoxia/HIFs, you can refer to our hypoxia-related FAQs: https://www.novusbio.com/support/hypoxia-and-hif-faqs
  11. Show All 14 FAQs.

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Blogs on HIF-1 alpha. Showing 1-10 of 37 blog posts - Show all blog posts.

Hypoxia-Dependent CAR Stabilizing Construct in T cells Improves Solid Tumor Targeting and Efficacy
By Victoria Osinski, PhDDespite advances in the development of cancer immunotherapies, those specifically targeting tumors still remains limited. Currently, there is great interest in utilizing chimeric antigen rece...  Read full blog post.

Tired T cells: Hypoxia Drives T cell Exhaustion in the Tumor Microenvironment
By Hunter MartinezThe paradigm shifting view of the immune system being leveraged to target cancer has led to numerous therapeutic breakthroughs. One major cell group responsible for this revelation is a T cell. ...  Read full blog post.

Understanding ‘Y’ in Breast Cancer: Crucial Role of DNA/RNA-binding Protein YB-1 in the Development, Pre-Invasive, and Metastatic Phases
Jamshed Arslan, Pharm D, PhD In the United States, 1 in 8 women will be diagnosed with breast cancer in her lifetime.1 Despite the prevalence, cancer genesis is a mystery. The heterogeneity of cancers makes it diff...  Read full blog post.


  Read full blog post.

Breast cancer stem cells survive chemotherapy through S100A10-ANXA2-SPT6 interaction that epigenetically promotes OCT4-mediated stemness
By Jamshed Arslan, Pharm D, PhDBreast cancer is the most common cancer among women that causes the greatest number of cancer-related deaths worldwide. After radiotherapy or cytotoxic chemotherapy like paclitax...  Read full blog post.

mTOR Signaling and the Tumor Microenvironment
By Yoskaly Lazo-Fernandez, PhD The mammalian target of rapamycin (mTOR) is a conserved serine/threonine kinase that, as a member of two distinct intracellular protein complexes, mTORC1 and mTORC2, regulates protein ...  Read full blog post.

Bad news for stomach cancer: BAMBI protein inhibits gastric carcinoma via TGF-beta/epithelial-mesenchymal transition signaling
By Jamshed Arslan Pharm.D. Gastric carcinoma is the second leading cause of cancer-related deaths worldwide. One of the key features of gastric carcinoma is acidosis, which promotes growth and metastasis of gastric ...  Read full blog post.

Developmental regulator Daam2 promotes glial cell tumors by degrading Von Hippel-Lindau protein
By Jamshed Arslan Pharm.D. Glioblastoma is an aggressive type of cancer that forms from the star-shaped glial cells of the central nervous system, called astrocytes. Intriguingly, several genes linked to glioblasto...  Read full blog post.

Stemness for Surviving Hypoxia: TGF-beta/Smad Signaling in Multiple Myeloma
By Jamshed Arslan Pharm.D. Multiple myeloma (MM) is a cancer of antibody-producing plasma cells. The bone marrow (BM) of MM patients is hypoxic, and MM cells overexpress many cancerous genes that are regulated by hy...  Read full blog post.

Forecasting and Targeting a Rare Cancer with Hypoxia-Inducible Factor
By Jamshed Arslan Pharm.D. Cancers of nerve, adipose, and other soft tissues are called soft tissue sarcomas (STS). Malignant peripheral nerve sheath tumor (MPNST) is an example of a rare and hard-to-treat STS; eve...  Read full blog post.

Showing 1-10 of 37 blog posts - Show all blog posts.
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Gene Symbol HIF1A
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