HIF-1 alpha Antibody - BSA Free

Images

 
Biological Strategies: ChIP-qPCR assay of HIF-1 binding to hypoxia response element (HRE) of the PKM2 gene in human MDA-MB-231 breast cancer cells treated with vehicle or DMOG (HIF inducer) for 8 hours. Negative ...read more
Normoxic pre-treatment sensitizes glioma cells to radiation after graded chronic hypoxic (GCH) exposure. The results of anchorage-independent colony forming assays are shown for U87, U87-luc, GL261 glioma cells and 0308 ...read more
Immunofluorescence of wounded tissue and normal unwounded skin. Representative images for immunofluorescent staining of HIF-1 alpha on healed wounded skin and adjacent normal unwounded skin of wild type, heterozygous ...read more
Staining in normal mouse kidney showing (A) staining with HIF-1 alpha antibody and (B) blocking peptide used at 10-fold excess concentration. Images provided by Dr Yves Heremans. Please see the product review of ...read more
Biological Strategies: Analysis of HIF-1 alpha in mouse hepatocytes (left lane: normoxic and right lane: hypoxic) using HIF-1 alpha antibody. Western blot image submitted by a verified customer review.
Analysis in paraffin section of mouse skin wound using NB100-479 at 1:6000. Image courtesy of product review by Ian Darby of RMIT University.
Immunofluorescence (IF) labeling and confocal microscopy analysis of the distribution of HIF-1alpha (A1-D1) and Abeta (A2-D2) in APPsw cells. DFO treatment significantly increased HIF-1alpha IF (D1) and reduced the ...read more
MDA-MB-231 cells were exposed to 20% or 1% O2 for 16 hours, and chromatin immunoprecipitation (ChIP) was performed with the indicated antibody (Ab). Primers flanking the HIF binding site in the S100A10 gene were used ...read more
Biological Strategies: Analysis in A. PC12 (rat) cells, 20% oxygen and B. PC12 (rat) cells, 1% oxygen.
Biological Strategies: Analysis in a hypoxic sample. Lane 1: CoCl2 treated Cos-7 nuclear extract (50 ug, hypoxic). Lane 2: Untreated Cos-7 nuclear extract (50 ug, normoxic). 10 second ECL exposure.
U251 cells exposed to 4 hour hypoxia (0.5% oxygen) incubation. Western blot image submitted by a verified customer review.
Detection of HIF-1 Alpha (green) in RCC4 cells using NB100-479. Nuclei (Blue) were counterstained using Hoechst 33258.
Analysis of HIF-1 alpha in human kidney tumor.
Staining of human placenta, villi.
Effects of different routes of CDDP administration on remaining viable tumor tissue. Representative images of tumor tissue (i.v. CDDP, i.a. CDDP) stained for HIF-1alpha (20X). Scale bar corresponds to 100 um. Image ...read more
Hif1alpha expression in preimplantation embryos. Co-staining of OCT4 and Hif1alpha in inner cell mass of blastocyst (top) and negative control images without primary antibodies (bottom). Scale bar: 30um. Image collected ...read more
Simple Western lane view shows lysates of MCF +/- DMOG, BioSpherix MSCs in hypoxic conditions, and BioSpherix MSCs in normoxic conditions loaded at 0.5 mg/ml. A band was detected for HIF-1 alpha at approximately 116 kDa ...read more
Cofilin phosphorylation is HIF-1 alpha dependent.Wt, shC, and the HIF-1 alpha knock down cell clones c1 and c2 cells were lysed after 24 hrs of normoxia (20% O2) or hypoxia (1% O2). Cell extracts were analysed by ...read more
Hif1alpha mediates Glut1 induction downstream of Bmp-mTORC1 signaling. d) Western blot analyses in primary chondrocytes in response to BMP2 with or without the Hif1alpha inhibitors for 24 h. Quantification of all ...read more
Effects of different routes of CDDP administration on remaining viable tumor tissue. B) Representative images of tumor tissue (i.v. CDDP, i.a. CDDP) stained for HIF-1alpha (20X). Scale bar corresponds to 100 um. Image ...read more
Involvement of HIF-1alpha in alveolar epithelial cell phenotypic changes induced by hypoxia. Primary rat AECs were cultured in normoxia (Nx) (21% O2) or hypoxia (Hx) (1.5% O2) during 6 days in presence or absence of 10 ...read more
Multiple distinct mechanisms for downregulation of AKT activity in TSC1/2-deficient SCs. A-F: Western blots of sciatic nerve lysates from control and TSC2-SCKO/TSC1-SCKO mice at age P28 (A-E) or P14 (F) (3 mice per ...read more
Hypoxia downregulates MHC class I expression via HIF transcription factors.(A-C): siRNA mediated knockdown of HIF-1alpha reversed hypoxic downregulation of MHC class I expression as compared with the scrambled, ...read more
Kidney-derived Sca-1+ cells produce Epo. (B) HIF-1alpha and HIF-2alpha protein levels in Sca-1+ kidney-derived cells after exposure to normoxia (20% O2) or hypoxia (1% O2). Numbers below the blots indicate the fold ...read more
G9a inhibition reduces cell motility in breast cancer cell lines. G9a inhibition represses breast cancer cell motility in vitro. (E) Western blot analysis of G9a, H3K9me1 and H3K9me2 in MDA-MB-231 transfected with ...read more
Effect of 24 h UO on renal HIF-1alpha/2alpha expression.WT mice were exposed to either sham-operation or left UO, which continued for 24 hours, and then was released. The left kidneys were harvested immediately (d0), 2, ...read more
ONECUT2 modulates HIF1alpha binding to chromatin in NE-like PC3 cells. a Western blot of HIF1alpha and ONECUT2 in PC3 cells under normoxic and hypoxic conditions. Two different siRNAs targeting ONECUT2 were mixed ...read more
Hypoxic markers accumulate in miR-29-sponge Zebrafish hearts. (A) Representative western blot analysis of hypoxia inducible factor 1alpha (HIF1alpha) expression in Wild Type and miR-29-sponge Zebrafish heart. In each ...read more
HIF-1alpha protein increases in HCT116 cells exposed to rapid intermittent hypoxia. Increased HIF-1alpha increases HIF-1 transactivation, leading to increased expression of HIF-1 target genes at both the mRNA and ...read more
Pharmacological inhibition of ABCC3 reduces cell proliferation through STAT3 and HIF1alpha dysregulation and induction of apoptosis. a) Representative Western blot images show the effects of pharmacological inhibition ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Ca, Fi, Gt, Ha, Pm, Rb, ZeSpecies Glossary
Applications WB, Simple Western, Flow, ICC/IF, IHC, IP, WB, ChIP, IHC
Clonality
Polyclonal
Host
Rabbit
Conjugate
Unconjugated
Format
BSA Free
Concentration
1.0 mg/ml
Validated by:
   

Biological Strategies

     

Order Details

HIF-1 alpha Antibody - BSA Free Summary

Immunogen
This HIF-1 alpha antibody was developed against a fusion protein including amino acids 530 - 825 of the mouse HIF-1 alpha protein [Uniprot# Q61221].
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 (ChIP) 1:10-1:500
  • Flow Cytometry 1:10 - 1:1000. Use reported in scientific literature (PMID 21917971)
  • Immunoblotting reported in scientific literature (PMID 28506759)
  • Immunocytochemistry/ Immunofluorescence 1:100
  • Immunohistochemistry Whole-Mount reported in scientific literature (PMID 27148974)
  • Immunohistochemistry 5 - 10 ug/mL
  • Immunohistochemistry-Frozen 5 - 10 ug/mL
  • Immunohistochemistry-Paraffin 5 - 10 ug/mL
  • Immunoprecipitation 1:10 - 1:500
  • Simple Western
  • Western Blot 1-2ug/ml
Application Notes
WB recognizes a band in hypoxic samples at ~115 kDa. HIF-1 alpha protein (NBC1-18422) has been successfully used for antibody preadsorption/blocking assay in IHC-P with HIF-1 alpha antibody NB100-479.
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 Pancreas Whole Tissue Lysate (Adult Whole Normal)
COS-7 Nuclear Hypoxic Induced Cell Lysate
Human Bone Marrow Whole Tissue Lysate (Adult Whole Diabetes)
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
Control Peptide
HIF-1 alpha Recombinant Protein (NBC1-18422)
Reviewed Applications
Read 28 Reviews rated 4.4
using
NB100-479 in the following applications:

Publications
Read Publications using
NB100-479 in the following applications:

Reactivity Notes

Use in Zebrafish reported in scientific literature (PMID:35457018).Use in Mouse reported in scientific literature (PMID:33727588). Use in Goat reported in scientific literature (PMID:21599540).

Packaging, Storage & Formulations

Storage
Store at -20C.
Buffer
PBS
Preservative
0.02% Sodium Azide
Concentration
1.0 mg/ml
Purity
Immunogen affinity purified

Alternate Names for HIF-1 alpha Antibody - BSA Free

  • 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 (NB100-479)(490)

We have publications tested in 11 confirmed species: Human, Mouse, Rat, Canine, Fish, Goat, Hamster, Moth - Trichoplusia ni (Cabbage Looper), Porcine, Rabbit, Zebrafish.

We have publications tested in 17 applications: B/N, ChIP, Chemotaxis, CyTof, EA, FLOW, IB, ICC/IF, IF/IHC, IHC, IHC-Fr, IHC-FrFl, IHC-P, IHC-WhMt, IP, In Vivo, WB.


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B/N
(3)
ChIP
(1)
Chemotaxis
(18)
CyTof
(1)
EA
(1)
FLOW
(3)
IB
(1)
ICC/IF
(38)
IF/IHC
(40)
IHC
(8)
IHC-Fr
(9)
IHC-FrFl
(1)
IHC-P
(42)
IHC-WhMt
(1)
IP
(12)
In Vivo
(2)
WB
(266)
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Human
(139)
Mouse
(190)
Rat
(29)
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(1)
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(1)
Goat
(1)
Hamster
(2)
Moth - Trichoplusia ni (Cabbage Looper)
(1)
Porcine
(1)
Rabbit
(2)
Zebrafish
(1)
All Species
Showing Publications 1 - 10 of 490. Show All 490 Publications.
Publications using NB100-479 Applications Species
Zhu X, Chen J, Li W et al. Hypoxia-responsive CAR-T cells exhibit reduced exhaustion and enhanced efficacy in solid tumors Cancer research 2023-10-24 [PMID: 37874330]
Yamazaki T, Mimura I, Kurata Y et al. Dznep, a histone modification inhibitor, inhibits HIF1? binding to TIMP2 gene and suppresses TIMP2 expression under hypoxia Physiol Rep 2023-09-14 [PMID: 37710084]
Maduabuchi, WO;Tansi, FL;Heller, R;Hilger, I; Hyperthermia Influences the Secretion Signature of Tumor Cells and Affects Endothelial Cell Sprouting Biomedicines 2023-08-12 [PMID: 37626752] (ICC/IF) ICC/IF
Kunke, M;Knöfler, H;Dahlke, E;Zanon Rodriguez, L;Böttner, M;Larionov, A;Saudenova, M;Ohrenschall, GM;Westermann, M;Porubsky, S;Bernardes, JP;Häsler, R;Magnin, JL;Koepsell, H;Jouret, F;Theilig, F; Targeted deletion of von-Hippel-Lindau in the proximal tubule conditions the kidney against early diabetic kidney disease Cell death & disease 2023-08-26 [PMID: 37626062]
Vellama H, Eskla KL, Eichelmann H et al. VHL-deficiency leads to reductive stress in renal cells Free radical biology & medicine 2023-07-26 [PMID: 37506952] (WB, Human)

Details:
1:1000 WB dilution
WB Human
Ouhaddi Y, Dalisson B, Rastinfard A et al. Necrosis reduction efficacy of subdermal biomaterial mediated oxygen delivery in ischemic skin flaps Biomaterials advances 2023-06-16 [PMID: 37392519] (IHC, Rat) IHC Rat
Dai X, Ruan J, Guo Y et al. Enhanced Radiotherapy Efficacy and Induced Anti-tumor Immunity in HCC by Improving Hypoxia Microenvironment using Oxygen Microcapsules Chemical Engineering Journal 2021-04-01 (IF/IHC, ICC/IF) IF/IHC, ICC/IF
Wang N, Su X, Sams D et al. P300/CBP Regulates HIF-1 Dependent Sympathetic Activation and Hypertension by Intermittent Hypoxia American journal of respiratory cell and molecular biology 2023-10-24 [PMID: 37874694]

Details:
DNA immunoprecipitation
Wang D, Zhu Z, Fu Y et al. Bromodomain-containing protein 4 activates androgen receptor transcription and promotes ovarian fibrosis in PCOS Cell reports 2023-09-04 [PMID: 37669164] (IP, Rat) IP Rat
Mawambo G, Oubaha M, Ichiyama Y et al. HIF1?-dependent hypoxia response in myeloid cells requires IRE1? Journal of neuroinflammation 2023-06-21 [PMID: 37344842] (WB, IP, Mouse) WB, IP Mouse
Show All 490 Publications.

Reviews for HIF-1 alpha Antibody (NB100-479) (28) 4.428

Average Rating: 4.4
(Based on 28 reviews)
We have 28 reviews tested in 6 species: Human, Mouse, Rat, Human and Human Breast Cancer Cell Lines, Human and Mouse, Other.

Reviews using NB100-479:
Filter by Applications
WB
(22)
ChIP
(2)
IHC-P
(4)
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Filter by Species
Human
(13)
Mouse
(9)
Rat
(2)
Human and Human Breast Cancer Cell Lines
(1)
Human and Mouse
(1)
Other
(2)
All Species
Images Ratings Applications Species Date Details
Western Blot HIF-1 alpha NB100-479
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5
reviewed by:
Verified Customer
WB Human 05/28/2021
View

Summary

ApplicationWestern Blot
Sample TestedA549 cells
SpeciesHuman
LotAQ-2
Western Blot HIF-1 alpha NB100-479
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5
reviewed by:
Verified Customer
WB Human 03/15/2021
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Summary

ApplicationWestern Blot
Sample TestedHep3B human hepatocellular carcinoma cell line
SpeciesHuman
LotAN-1
Western Blot HIF-1 alpha NB100-479
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5
reviewed by:
Yongkang Yang
WB Human and Mouse 10/10/2020
View

Summary

ApplicationWestern Blot
Sample Testedcell line, whole cell lysate
SpeciesHuman and Mouse
LotAP-2
Chromatin Immunoprecipitation HIF-1 alpha NB100-479
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5
reviewed by:
Verified Customer
ChIP Human and Human Breast Cancer Cell Lines 05/17/2020
View

Summary

ApplicationChromatin Immunoprecipitation
Sample TestedBreast cancer cells
SpeciesHuman and Human Breast Cancer Cell Lines
LotAN-1
Western Blot HIF-1 alpha NB100-479
Enlarge
3
reviewed by:
Verified Customer
WB Human 04/16/2019
View

Summary

ApplicationWestern Blot
Sample Tested16-HBE
SpeciesHuman
LotAL-6
Western Blot HIF-1 alpha NB100-479
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5
reviewed by:
Verified Customer
WB Human 09/18/2018
View

Summary

ApplicationWestern Blot
Sample TestedHuman Cancer Cells
SpeciesHuman
LotAN-1
Immunohistochemistry-Paraffin HIF-1 alpha NB100-479
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5
reviewed by:
Verified Customer
IHC-P Mouse 10/25/2017
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Summary

ApplicationImmunohistochemistry-Paraffin
Sample TestedIHC on parrafin sections (mouse)
SpeciesMouse
LotAK-4
Western Blot HIF-1 alpha NB100-479
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4
reviewed by:
Kristina Cook
WB Human 02/23/2017
View

Summary

ApplicationWestern Blot
Sample TestedU251 glioma cell line; whole cell lysate
SpeciesHuman

Comments

Comments4 hour hypoxia (0.5% oxygen) incubation
  5
reviewed by:
Verified Customer
WB Mouse 09/21/2016
View

Summary

ApplicationWestern Blot
Sample TestedMouse kidney
SpeciesMouse
Western Blot HIF-1 alpha NB100-479
Enlarge
4
reviewed by:
Verified Customer
WB Mouse 08/06/2016
View

Summary

ApplicationWestern Blot
SpeciesMouse
Western Blot HIF-1 alpha NB100-479
Enlarge
5
reviewed by:
Zhongxiao Wang
WB Mouse 10/05/2015
View

Summary

ApplicationWestern Blot
Sample Testedmouse retina
SpeciesMouse
LotNB100-479
  3
reviewed by:
Verified Customer
WB Human 08/23/2015
View

Summary

ApplicationWestern Blot
Sample TestedHuman cancer cell whole cell lysate
SpeciesHuman
  4
reviewed by:
Verified Customer
WB Human 03/27/2015
View

Summary

ApplicationWestern Blot
Sample TestedHela lysates treated with MG132
SpeciesHuman
Western Blot HIF-1 alpha NB100-479
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4
reviewed by:
Verified Customer
WB Mouse 02/18/2015
View

Summary

ApplicationWestern Blot
Sample TestedMouse heart
SpeciesMouse
LotJ1
  5
reviewed by:
Verified Customer
ChIP Human 12/15/2014
View

Summary

ApplicationChromatin Immunoprecipitation
Sample TestedHEK293
SpeciesHuman
LotAA-1
  5
reviewed by:
Verified Customer
WB Mouse 12/12/2014
View

Summary

ApplicationWestern Blot
Sample TestedSee PMID 23232625
SpeciesMouse
  4
reviewed by:
Verified Customer
WB Other 09/15/2014
View

Summary

ApplicationWestern Blot
Sample TestedSK-N-BE(2) whole cell lysate
SpeciesOther
LotAA-1
Western Blot HIF-1 alpha NB100-479
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4
reviewed by:
Xiuquan Luo
WB Human 09/12/2014
View

Summary

ApplicationWestern Blot
Sample TestedHEK293 cell lysate
SpeciesHuman
  4
reviewed by:
Verified Customer
WB Other 08/18/2014
View

Summary

ApplicationWestern Blot
Sample TestedPC12 whole cell lysate
SpeciesOther
LotAA-1
Western Blot HIF-1 alpha NB100-479
Enlarge
4
reviewed by:
Ming Lu
WB Human 08/15/2014
View

Summary

ApplicationWestern Blot
Sample TestedHuman Pancreatic Cancer Cell line
SpeciesHuman
LotAA
Western Blot HIF-1 alpha NB100-479
Enlarge
4
reviewed by:
Molly Jenkins
WB Human 07/18/2014
View

Summary

ApplicationWestern Blot
Sample TestedNuclear fraction, Human cell line
SpeciesHuman
Western Blot HIF-1 alpha NB100-479
Enlarge
4
reviewed by:
Jason Lee
WB Human 10/29/2013
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Summary

ApplicationWestern Blot
Sample TestedHuman MCF7 breast epithelial cells
SpeciesHuman
LotLot X-1
Immunohistochemistry-Paraffin HIF-1 alpha NB100-479
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5
reviewed by:
Verified Customer
IHC-P Mouse 06/27/2013
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Summary

ApplicationImmunohistochemistry-Paraffin
Sample Testedparaffin section of skin wound
SpeciesMouse
Western Blot HIF-1 alpha NB100-479
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5
reviewed by:
Sandhya Khurana
WB Rat 02/05/2012
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Summary

ApplicationWestern Blot
Sample TestedPC12 nuclear extract, Sample Amount: 50ug
SpeciesRat
  5
reviewed by:
Verified Customer
WB Rat 01/03/2012
View

Summary

ApplicationWestern Blot
Sample TestedPC12 cell whole lysate, Sample Amount: 25ug
SpeciesRat
LotP1
Immunohistochemistry-Paraffin HIF-1 alpha NB100-479
Enlarge
4
reviewed by:
Verified Customer
IHC-P Mouse 07/30/2011
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Summary

ApplicationImmunohistochemistry-Paraffin
Sample Testednormal mouse kidney
SpeciesMouse
Immunohistochemistry-Paraffin HIF-1 alpha NB100-479
Enlarge
5
reviewed by:
Paramahamsa Maturu
IHC-P Human 03/01/2011
View

Summary

ApplicationImmunohistochemistry-Paraffin
Sample TestedHuman Kidney tumor
SpeciesHuman
LotI
Western Blot HIF-1 alpha NB100-479
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4
reviewed by:
JORCYK LAB
WB Mouse 02/01/2011
View

Summary

ApplicationWestern Blot
Sample Tested4T1 cells whole cell lysate
SpeciesMouse

Product General Protocols

View specific protocols for HIF-1 alpha Antibody (NB100-479): Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

Video Protocols

WB Video Protocol
ICC/IF Video Protocol

FAQs for HIF-1 alpha Antibody (NB100-479). (Showing 1 - 10 of 17 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 (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.Also, 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.
  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 (5)    Last but not the least, Novus technical support team may be contacted via email
  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.
  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.
  11. Show All 17 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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Bioinformatics

Gene Symbol HIF1A
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