HIF-1 alpha Antibody (H1alpha67)

Images

 
Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Analysis of HIF-1 alpha in human hepatocytes from cancer patient using anti-HIF-1 alpha antibody. Image from verified customer review.
Immunocytochemistry/ Immunofluorescence: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Expression of HIF-1 in intresititial pig cells under 12% of 02 condition. Image from verified customer review.
Immunohistochemistry-Paraffin: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Analysis using the Biotin conjugate of NB100-105. Staining of human glioblastoma multiforme.
Flow Cytometry: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Analysis using the Alexa Fluor (R) 488 conjugate of NB100-105. Staining of HIF-1 alpha in multiple myeloma cells: H929 cells (0.5x10^6) were stained with ...read more
Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - HIF-1 alpha induction by CoCl2 on Caki-1 cell lysate. Image from verified customer review.
Western Blot: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Analysis using the HRP conjugate of NB100-105. Detection of 50ug cobalt chloride induced COS-7 nuclear extracts (NB800-PC26) using NB100-105.
Immunohistochemistry: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Staining of HIF1 alpha in human kidney. Renal tubular epithelium showed moderate membranous, cytoplasmic and nuclear staining, and glomeruli showed ...read more
Flow Cytometry: HIF-1 alpha Antibody (H1alpha67) [NB100-105] - Analysis using the Alexa Fluor (R) 488 conjugate of NB100-105. Staining of HIF-1 alpha in H929 cells using HIF-1 alpha antibody. Image from verified ...read more

Product Details

Summary
Reactivity Hu, Mu, Rt, Po, Bv, Ft, Mk, Pm, Rb, Sh, Xp, YeSpecies Glossary
Applications WB, ChIP, ELISA, Flow, GS, ICC/IF, IHC, IHC-Fr, IHC-P, IP, LA, TCS
Clone
H1alpha67
Clonality
Monoclonal
Host
Mouse
Concentration
1.0 mg/ml

HIF-1 alpha Antibody (H1alpha67) Summary

Immunogen
A fusion protein containing amino acids 432 - 528 of human HIF-1 alpha [UniProt# Q16665].
Localization
HIF-1 is a nuclear protein
Isotype
IgG2b
Clonality
Monoclonal
Host
Mouse
Gene
HIF1A
Purity
Protein G purified
Innovator's Reward
Test in a species/application not listed above to receive a full credit towards a future purchase.

Packaging, Storage & Formulations

Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Conjugate
Unconjugated
Buffer
PBS with 1% BSA
Preservative
0.05% Sodium Azide
Concentration
1.0 mg/ml
Purity
Protein G purified

Applications/Dilutions

Dilutions
  • Western Blot 1:500
  • Chromatin Immunoprecipitation 1 - 5 ug/IP
  • ELISA 1:100 - 1:2000
  • Flow Cytometry 1:10 - 1:1000
  • Gel Super Shift Assays 1:1 - 1:100
  • Immunocytochemistry/Immunofluorescence 1:50
  • Immunohistochemistry 1:20 - 1:50
  • Immunohistochemistry-Frozen 1:20 - 1:50
  • Immunohistochemistry-Paraffin 1:20 - 1:50
  • Immunoprecipitation 1:10 - 1:500
  • Tissue Culture Substratum
  • Ligand Activation
Application Notes
ChIP usage was reported in scientific literature. ELISA usage was reported in scientific literature (PMID: 20042684). Gel Super Shift Assays usage was reported in scientif literature (PMID: 22411794). Ligand Activation usage was reported in scientific literature (PMID: 26147748). IHC-P: No detection of Mouse is reported. WB: Detection of Mouse protein. In WB, a band can be seen at 120 kDa representing HIF-1 alpha in induced tissues and cells. Multiple bands may be seen at 100-120 kDa representing post-translational modification of HIF-1 alpha. For WB, testing on nuclear extracts is recommended. 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.
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.
Reviewed Applications
Read 30 Reviews rated 4.2
using
NB100-105 in the following applications:

Publications
Read Publications using
NB100-105 in the following applications:

  • 39 publications
  • 5 publications
  • 3 publications
  • 2 publications
  • GS
    7 publications
  • IA
    1 publication
  • 48 publications
  • IHC
    91 publications
  • 4 publications
  • 78 publications
  • IP
    26 publications
  • WB
    334 publications

Reactivity Notes

Candida Albicans reactivity was reported by customer review. Xenopus reactivity was reported in scientific literature (PMID: 18303027). Please note that this antibody is reactive to Mouse and derived from the same host, Mouse. Additional Mouse on Mouse blocking steps may be required for IHC and ICC experiments. Please contact Technical Support for more information.

Alternate Names for HIF-1 alpha Antibody (H1alpha67)

  • anti-HIF-1 alpha
  • anti-HIF1A
  • ARNT-interacting protein
  • Basic-helix-loop-helix-PAS protein MOP1
  • BHLHE78
  • Class E basic helix-loop-helix protein 78
  • H1alpha67
  • HIF1 alpha
  • HIF-1 alpha
  • HIF1A
  • HIF-1-alpha
  • HIF1-alpha
  • hypoxia inducible factor 1, alpha subunit (basic helix-loop-helix transcriptionfactor)
  • hypoxia-inducible factor 1-alpha
  • Member of PAS protein 1
  • member of PAS superfamily 1
  • MOP1HIF1-ALPHA
  • PAS domain-containing protein 8
  • PASD8alpha subunit (basic helix-loop-helix transcriptionfactor)

Background

HIF1 (hypoxia-inducible factor 1), a heterodimeric transcription factor complex central to cellular response to hypoxia, consists of two subunits (HIF-1 alpha and HIF-1 beta) which are basic helix-loop-helix proteins of the PAS (Per, ARNT, Sim) family. Expression of HIF-1 alpha protein is regulated by cellular oxygen level alterations as well as in oxygen-independent manner via different cytokines (through the PI3K-AKT-mTOR pathway), growth factors, oncogenic activation, or loss of tumor suppressor function etc. In normoxic cells, HIF-1 alpha is proline hydroxylated leading to a conformational change that promotes its binding to the VHL (von Hippel Lindau) protein E3 ligase complex; ubiquitination and followed by rapid proteasomal degradation. Hypoxia as well as chemical hydroxylase inhibitors (desferrioxamine, cobalt etc.) inhibit HIF-1 alpha degradation and lead to its accumulation in the cells, whereas, contrastingly, HIF-1 beta/ARNT (AhR nuclear translocator) remains stable under both conditions. Besides their critical role in hypoxic response, HIF1s regulates the transcription of genes responsible for angiogenesis, erythropoiesis/iron-metabolism, glucose metabolism, cell proliferation/survival, adipogenesis, carotid body formation, B lymphocyte development and immune reactions.

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.

Customers Who Viewed This Item Also Viewed...

NBP1-89843
Species: Hu
Applications: ICC/IF, IHC, IHC-P
NB100-124
Species: Hu, Mu, Rt, Bv, Ft, Pm, Sh
Applications: WB, ChIP, GS, ICC/IF, IHC, IHC-P, IP
NB100-485
Species: Hu
Applications: WB, IP
NB100-122
Species: Hu, Mu, Rt, Fi, Ha, Pm, Sh
Applications: WB, Simple Western, ChIP, ELISA, Flow, GS, ICC/IF, IHC, IHC-Fr, IHC-P, IP
MAB2055
Species: Hu, Mu, Rt
Applications: WB, Simple Western, ICC, ICFlow
NB110-39113
Species: Hu, Mu, Rt
Applications: WB, ChIP, Flow, ICC/IF, IHC, IHC-Fr, IHC-P
NB100-417
Species: Hu, Mu, Rt, Ca, Pl
Applications: WB, Simple Western, ELISA, Flow, GS, ICC/IF, IHC, IHC-Fr, IHC-P, ChIP
MAB1489
Species: Hu
Applications: WB
MAB18251
Species: Hu, Mu
Applications: WB, Simple Western
NBP1-47842
Species: Hu, Rt, Ca, Mk
Applications: WB, IHC, IHC-P, IP
NBP1-19855
Species: Hu, Mu, Rt
Applications: WB, ICC/IF, IHC, IHC-P
NB100-105
Species: Hu, Mu, Rt, Po, Bv, Ft, Mk, Pm, Rb, Sh, Xp, Ye
Applications: WB, ChIP, ELISA, Flow, GS, ICC/IF, IHC, IHC-Fr, IHC-P, IP, TCS, LA
NB300-605
Species: Hu, Mu, Rt, Po, Av, Fe, Fi, Rb, Xp
Applications: WB, ICC/IF, IHC, IHC-P
NB100-137
Species: Hu, Mu, Rt, Pm
Applications: WB, Simple Western, Flow, ICC/IF, IHC, IHC-P, IP

Publications for HIF-1 alpha Antibody (NB100-105)(614)

We have publications tested in 10 confirmed species: Human, Mouse, Rat, Bovine, Canine, Porcine, Primate, Rabbit, Sheep, Xenopus.

We have publications tested in 12 applications: ChIP, ELISA, EMSA, FLOW, GS, IA, ICC/IF, IHC, IHC-Fr, IHC-P, IP, WB.


Filter By Application
ChIP
(39)
ELISA
(5)
EMSA
(3)
FLOW
(2)
GS
(7)
IA
(1)
ICC/IF
(48)
IHC
(91)
IHC-Fr
(4)
IHC-P
(78)
IP
(26)
WB
(334)
All Applications
Filter By Species
Human
(303)
Mouse
(165)
Rat
(91)
Bovine
(3)
Canine
(1)
Porcine
(4)
Primate
(2)
Rabbit
(4)
Sheep
(2)
Xenopus
(1)
All Species
Showing Publications 1 - 10 of 614. Show All 614 Publications.
Publications using NB100-105 Applications Species
Soncini M, Corna G, Moresco M et al. 24-Hydroxycholesterol participates in pancreatic neuroendocrine tumor development Proc. Natl. Acad. Sci. U.S.A. Sep 26 2016 12:00AM [PMID:27671648] (ChIP, Mouse) ChIP Mouse
Choi YK, Maki T, Mandeville ET et al. Dual effects of carbon monoxide on pericytes and neurogenesis in traumatic brain injury Nat. Med. Sep 26 2016 12:00AM [PMID:27668935] (IHC, Mouse) IHC Mouse
Maugeri G, D'Amico AG, Saccone S et al. PACAP and VIP Inhibit HIF-1alpha-Mediated VEGF Expression in a Model of Diabetic Macular Edema J. Cell. Physiol. Sep 23 2016 12:00AM [PMID:27661459] (WB, Human) WB Human
Miyake T, Nakamura S, Zhao M et al. Cold sensitivity of TRPA1 is unveiled by the prolyl hydroxylation blockade-induced sensitization to ROS Nat Commun Sep 16 2016 12:00AM [PMID:27628562] (WB, Human) WB Human
Budda SA, Girton A, Henderson JG, Zenewicz LA. Transcription Factor HIF-1alpha Controls Expression of the Cytokine IL-22 in CD4 T Cells. J. Immunol. Aug 17 2016 12:00AM [PMID:27534553] (WB) WB
Nagamine Y, Tojo K, Yazawa T et al. Inhibition of Prolyl Hydroxylase Attenuates Fas Ligand-induced Apoptosis and Lung Injury in Mice. Am J Respir Cell Mol Biol. Aug 5 2016 12:00AM [PMID:27494234]
Littlewood-Evans A, Sarret S, Apfel V et al. GPR91 senses extracellular succinate released from inflammatory macrophages and exacerbates rheumatoid arthritis. J. Exp. Med. Aug 22 2016 12:00AM [PMID:27481132] (WB, Mouse) WB Mouse
Zhang Q, Doucet M, Tomlinson RE et al. The hypoxia-inducible factor-1a activates ectopic production of fibroblast growth factor 23 in tumor-induced osteomalacia Bone Res 2016 Jul 29 [PMID:27468359]
Cui J, Duan B, Zhao X et al. MBD3 mediates epigenetic regulation on EPAS1 promoter in cancer Tumour Biol. 2016 Jul 27 [PMID:27465550] (WB, Sheep) WB Sheep
Stei MM, Loeffler KU, Kurts C et al. Impact of macrophages on tumor growth characteristics in a murine ocular tumor model Exp. Eye Res. 2016 Jul 15 [PMID:27426931] (IHC, Human) IHC Human
Show All 614 Publications.

Reviews for HIF-1 alpha Antibody (NB100-105) (30) 4.230

Average Rating: 4.2
(Based on 30 reviews)
We have 30 reviews tested in 4 species: Human, Mouse, Rat, Other.
We have 30 reviews tested in 4 application: WB, IHC, IF, ICC.

Reviews using NB100-105:
Filter by Applications
WB
(25)
IHC
(1)
IF
(2)
ICC
(2)
All Applications
Filter by Species
Human
(13)
Mouse
(11)
Rat
(3)
Other
(3)
All Species
Images Ratings Applications Species Date Details
Western Blot HIF-1 alpha NB100-105
Enlarge
reviewed by:
michael dodd
WB Mouse 08/17/2016
View

Summary

ApplicationWestern Blot
Sample TestedHL-1 cardiomyocytes
SpeciesMouse

Blocking

Blocking Details5% milk, in TBST for 1 hour at room temperature

Primary Anitbody

Dilution Ratio1 in 500, Overnight at 4C, in 5% milk with TBST

Secondary Antibody

Secondary DescriptionGoat anti-mouse, HRP
Secondary Concentration1 in 2000

Details

Detection NotesECL Prime, Licor scanner, 12 min exposure
 
reviewed by:
Anonymous
WB Human 04/18/2016
View

Summary

ApplicationWestern Blot
Sample TestedHEK293A whole cell lysate
SpeciesHuman
LotAW-1

Blocking

Blocking Details4% milk in TBST, 1hr at room temperature

Primary Anitbody

Dilution Ratio1:2000 dilution in TBST buffer, incubated at 4 degrees overnight (~16hrs)

Secondary Antibody

Secondary DescriptionGoat anti-mouse IgG
Secondary Manufacturer Cat#ThermoScientific #31430
Secondary Concentration1:7500

Details

Detection NotesBio-Rad Versadoc, exposure time was one image every 15s for 5 minutes, negative control was empty plasmid
Western Blot HIF-1 alpha NB100-105
Enlarge
reviewed by:
MENG xing
WB Mouse 03/24/2016
View

Summary

ApplicationWestern Blot
SpeciesMouse
LotBD-10
Special ApplicationsBone marrow cells
FileView PDF
Western Blot HIF-1 alpha NB100-105
Enlarge
reviewed by:
Anonymous
WB Mouse 10/07/2015
View

Summary

ApplicationWestern Blot
Sample TestedCell lysate
SpeciesMouse

Blocking

Blocking DetailsMilk

Primary Anitbody

Dilution RatioOvernight 1:500

Secondary Antibody

Secondary Descriptionanti-rabbit

Details

Detection NotesECL (doesn't work on Licor)
Western Blot HIF-1 alpha NB100-105
Enlarge
reviewed by:
Zhongxiao Wang
WB Mouse 10/05/2015
View

Summary

ApplicationWestern Blot
Sample Testedmouse retina
SpeciesMouse
LotNB100-479

Blocking

Blocking Details5%milk/T-BST buffer, room temparature, 2hr

Primary Anitbody

Dilution Ratio1:1000 in 5%milk/T-BST buffer, 4 degree, overnight

Secondary Antibody

Secondary DescriptionGoat anti-rabbit IgG, HRP
Secondary Manufacturer Cat#Sigma AQ132P
Secondary Concentration1:3000

Details

Detection NotesDevelop machine, 30s exposure shows clear band

Comments

CommentsThere's a non-specific band above for mouse retina samples
 
reviewed by:
Anonymous
WB Human 08/23/2015
View

Summary

ApplicationWestern Blot
Sample TestedHuman cancer cell whole cell lysate
SpeciesHuman

Blocking

Blocking DetailsBlocking with 5% non-fat milk for 30 min

Primary Anitbody

Dilution Ratio1:500, overnight

Secondary Antibody

Secondary DescriptionSheep anti-mouse IgG, HRP
Secondary Manufacturer Cat#GE Healthcare, NA931V
Secondary Concentration1:2000

Details

Detection NotesRegular Protocol
Western Blot HIF-1 alpha NB100-105
Enlarge
reviewed by:
Anonymous
WB Human 08/10/2015
View

Summary

ApplicationWestern Blot
Sample TestedHuman prostate cancer cell line protein samples
SpeciesHuman

Blocking

Blocking Details5% milk in TBS buffer supplemented with 0.1% tween20 at room temperature for one hour

Primary Anitbody

Dilution Ratio1:1000

Secondary Antibody

Secondary DescriptionAnti-Rabbit secondary diluted at 1:3000 in 5% milk
Secondary Manufacturer Cat#SC-2005
Secondary Concentration0.4 mg/ml

Details

Detection NotesWestern blotting using ECL reagent (purchased from GE life sciences) and x-ray films.
Western Blot HIF-1 alpha NB100-105
Enlarge
reviewed by:
Anonymous
WB Human 04/28/2015
View

Summary

ApplicationWestern Blot
SpeciesHuman
Special Applicationshuman hepatocytes from cancer patient
FileView PDF
 
reviewed by:
Anonymous
WB Mouse 12/17/2014
View

Summary

ApplicationWestern Blot
Sample TestedSee PIMD: 23148210
SpeciesMouse

Blocking

Blocking DetailsSee PIMD: 23148210

Details

Detection NotesSee PIMD: 23148210

Comments

CommentsPublished in PMID: 23148210
Western Blot HIF-1 alpha NB100-105
Enlarge
reviewed by:
Xiuquan Luo
WB Human 07/18/2014
View

Summary

ApplicationWestern Blot
Sample TestedCaki-1 cell lysate
SpeciesHuman

Blocking

Blocking Details1xPBST with 5% non-fat milk, RT 1 hour

Primary Anitbody

Dilution Ratio1xpBST with 5% non-fat milk, overnight 4C, 1:1000 dilution

Secondary Antibody

Secondary DescriptionGoat anti-mouse IgG-HRP
Secondary Manufacturer Cat#sc-2005
Secondary Concentration1:2000 dilution

Details

Detection NotesECL, exposured for 5 min
 
reviewed by:
Anonymous
WB Human 07/01/2014
View

Summary

ApplicationWestern Blot
Sample TestedHela Cell Lysate
SpeciesHuman
Western Blot HIF-1 alpha NB100-105
Enlarge
reviewed by:
Jessica Marinello
WB Human 05/09/2014
View

Summary

ApplicationWestern Blot
Sample TestedHela whole cell lysate
SpeciesHuman

Blocking

Blocking DetailsTBS milk 5% for 1 hour room temperature

Primary Anitbody

Dilution RatioAntibody diluition 1:1000 in TBS milk 5%, incubation time 2 hours room temperature

Secondary Antibody

Secondary DescriptionGoat anti-mouse IgG, HRP
Secondary Manufacturer Cat#santa cruz

Details

Detection NotesECL plus Western Blotting Detection Kit 5 minutes incubation room temperature, Phosphorimager detection system,
Western Blot HIF-1 alpha NB100-105
Enlarge
reviewed by:
Anonymous
WB Mouse 04/28/2014
View

Summary

ApplicationWestern Blot
Sample Testedmouse brain lysate
SpeciesMouse

Blocking

Blocking Details7% Milk 30 min RT

Primary Anitbody

Dilution Ratio1: 1000, 4 degree overnight in TBST

Secondary Antibody

Secondary DescriptionGoat anti-mouse HRP
Secondary Manufacturer Cat#NBP1-47596
Secondary Concentration1ug/mL
Western Blot HIF-1 alpha NB100-105
Enlarge
reviewed by:
Lindsy Dickerson
WB Mouse 04/28/2014
View

Summary

ApplicationWestern Blot
Sample TestedMouse
SpeciesMouse
Immunohistochemistry HIF-1 alpha NB100-105
Enlarge
reviewed by:
Ewa Kaniewska
IHC Other 04/14/2014
View

Summary

ApplicationImmunohistochemistry
SpeciesOther
FileView PDF
Immunofluorescence HIF-1 alpha NB100-105
Enlarge
reviewed by:
Ewa Kaniewska
IF Other 04/14/2014
View

Summary

ApplicationImmunofluorescence
SpeciesOther
FileView PDF
Western Blot HIF-1 alpha NB100-105
Enlarge
reviewed by:
Darlah Lopez
WB Mouse 03/28/2014
View

Summary

ApplicationWestern Blot
SpeciesMouse
Special ApplicationsPrimary 1:1000Secondary 1: 10000
FileView PDF
Western Blot HIF-1 alpha NB100-105
Enlarge
reviewed by:
Hua Li
WB Human 08/23/2012
View

Summary

ApplicationWestern Blot
Sample TestedWhole cell lysate
SpeciesHuman
LotAB7

Blocking

Blocking Details5% milk in TBS-T buffer, room temperature for one hours

Primary Anitbody

Dilution Ratio1:500 in 5% milk -TBS-T solution, 4C for overnight

Secondary Antibody

Secondary Descriptiongoat anti-mouse IgG, HRP
Secondary Manufacturer Cat#Santa Cruz, Cat# sc-2005
Secondary Concentration1:4000

Details

Detection NotesECL solution for 5min, then exposure to film 3-5min
Immunofluorescence HIF-1 alpha NB100-105
Enlarge
reviewed by:
Anonymous
IF Rat 08/22/2012
View

Summary

ApplicationImmunofluorescence
Sample TestedRAT KIDNEY
SpeciesRat
LotF2
CommentsPMID 21270095

Blocking

Blocking Detailsserum blocking for 30 minutes.

Primary Anitbody

Dilution Ratio1:100, overnight incubation,room temperature.

Details

Detection Notes1:1000 Alexa Flour 488, 3:1000 Hoechst 33342.
Fixation Details70% acetone 30% methanol.
Wash Description1xPBS, 5 minutes each,3 times.

Comments

CommentsPMID 21270095
Western Blot HIF-1 alpha NB100-105
Enlarge
reviewed by:
Camilla Sampaoli
WB Human 08/22/2012
View

Summary

ApplicationWestern Blot
Sample Testedhuman adrenal cancer cells (whole cell lysate)
SpeciesHuman

Blocking

Blocking Details5% non fat dry milk in PBS 1x + 0,1% tween; 1 h at RT

Primary Anitbody

Dilution Ratio1:500; 5% non fat dry milk in PBS 1x + 0,1% tween; o.n. at +4°C

Secondary Antibody

Secondary Descriptiongoat anti-rabbit HRP conjugated
Secondary Manufacturer Cat#Sigma-Aldrich
Secondary Concentration1:4000; 1 h at RT

Details

Detection Notesenhanced chemiluminescence (ECL) assay; exposure: 30 min
Western Blot HIF-1 alpha NB100-105
Enlarge
reviewed by:
Haofeng Ji
WB Mouse 05/15/2012
View

Summary

ApplicationWestern Blot
Sample TestedCell lysate
SpeciesMouse

Blocking

Blocking Details10% milk

Primary Anitbody

Dilution Ratio1:500

Secondary Antibody

Secondary Descriptionanti-rat IgG
Secondary Concentration1:2000

Details

Detection NotesECL
Western Blot HIF-1 alpha NB100-105
Enlarge
reviewed by:
Claudia Di Serio
WB Human 05/11/2012
View

Summary

ApplicationWestern Blot
Sample TestedHuman prostate cancer cells
SpeciesHuman
LotAB1
CommentsA good result of Novus' HIF-1 alpha Antibody (NB100-105) was obtained when we used a cell lysate with a plate (10 cm) of LNCaP. The cell line and the exposure time of plate was 20 minutes. We realized an immunoreactive band of about 120 KDa.

Blocking

Blocking Details5% dry milk inTBS 1X/tween (0.1%) buffer

Primary Anitbody

Dilution Ratio1:500, 4 Celcius, O/N

Secondary Antibody

Secondary Descriptionanti-mouse HRP conjugated
Secondary Manufacturer Cat#Pierce Biotechnology
Secondary Concentration1:2000

Details

Detection NotesImmunoreactive bands were visualized by enhanced chemiluminescence (ECL) assay

Comments

CommentsA good result of Novus' HIF-1 alpha Antibody (NB100-105) was obtained when we used a cell lysate with a plate (10 cm) of LNCaP. The cell line and the exposure time of plate was 20 minutes. We realized an immunoreactive band of about 120 KDa.
Western Blot HIF-1 alpha NB100-105
Enlarge
reviewed by:
Anonymous
WB Rat 12/14/2011
View

Summary

ApplicationWestern Blot
Sample TestedRat liver
SpeciesRat

Blocking

Blocking DetailsBlocking Buffer: 5% non-fat milk in TBST

Primary Anitbody

Dilution RatioDilution Ratio: 1:500, Incubation Time: overnight, Incubation Temp: 4 degrees Celsius

Secondary Antibody

Secondary DescriptionSecondary Ab: anti-mouse, Secondary Ab Dilution Ratio: 1:10000

Details

Detection NotesDetection Method: ECL
Western Blot HIF-1 alpha NB100-105
Enlarge
reviewed by:
Anonymous
WB Mouse 12/09/2011
View

Summary

ApplicationWestern Blot
Sample TestedMouse auditory cultured cell, Sample Amount: 30ug
SpeciesMouse
CommentsHIF-1 alpha degradation was quick, and our result was still detected by your antibody.

Blocking

Blocking DetailsBlocking Buffer: 1% BSA

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1:1000, Primary Ab Incubation Time: 1 hour

Secondary Antibody

Secondary DescriptionSecondary Ab: anti-mouse HRP

Details

Detection NotesDetection Method: Millipore ECL

Comments

CommentsHIF-1 alpha degradation was quick, and our result was still detected by your antibody.
 
reviewed by:
Anonymous
WB Human 11/21/2011
View

Summary

ApplicationWestern Blot
Sample TestedHuman
SpeciesHuman

Blocking

Blocking DetailsBlocking Buffer: 5% non-fat milk in TBST

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1:500, Primary Ab Incubation Time: overnight

Secondary Antibody

Secondary DescriptionSecondary Ab: anti-mouse

Details

Detection NotesDetection Method: Wesern Lighting plus ECL
 
reviewed by:
Lu-Hai Wang
ICC Human 11/15/2011
View

Summary

ApplicationImmunocytochemistry
Sample TestedHuman tissues
SpeciesHuman
CommentsIt's specific, but not very intense under current expected conditions.

Blocking

Blocking DetailsBlocking Buffer: BSA

Primary Anitbody

Dilution RatioPrimary Ab Dilution Ratio: 1:2000, Primary Ab Incubation Time: overnight

Secondary Antibody

Secondary DescriptionSecondary Ab: Goat anti-mouse IgG [Alexa Flour 594]

Details

Detection NotesDetection Visualization Method: Fluorescence microscopy

Comments

CommentsIt's specific, but not very intense under current expected conditions.
 
reviewed by:
Anonymous
WB Human 09/30/2011
View

Summary

ApplicationWestern Blot
Sample TestedMCF-7, Sample Amount: 100ug
SpeciesHuman
CommentsHIF-1 alpha has a half-life of approximately 1-5 min in normoxic conditions, so sometimes the failure to detect its protein is due to the postpone during the sample collection. I strongly suggest all the procedures completed entirely on ice and quickest lysis, processing and storage. Even though, this good antibody is still highly recommended. Cells from MCF-7 grown under normoxia or hypoxia were lysed for HIF-1 alpha blotting. For the best signal, we suggest higher amount of protein (80-120ug) and traditional tank (wet) electroblotting protocol. 10v overnight transferring is the longest but most efficient way to transfer big proteins as Hif-1a. 1:500 dilution of this Ab and 1:2000 secondary Ab work well.
Pub Med ID21193031

Blocking

Blocking DetailsBlocking Buffer: 5% Milk in TBST, Blocking Time: 2 hours, Blocking Temp: 20 degrees Celsius

Primary Anitbody

Dilution RatioDilution Ratio: 1:500, Incubation Dilution Buffer: 5% milk TBST, Incubation Time: 16 hours, Incubation Temp: 4 C

Secondary Antibody

Secondary DescriptionSecondary Ab: HRP labeled anti-mouse ab, Secondary Ab Dilution Ratio: 1:2000
Secondary Manufacturer Cat#7076 from cell signaling

Details

Detection NotesDetection Method: HRP, Exposure Time: 1 minute, Specific Bands: 90 kDa

Comments

CommentsHIF-1 alpha has a half-life of approximately 1-5 min in normoxic conditions, so sometimes the failure to detect its protein is due to the postpone during the sample collection. I strongly suggest all the procedures completed entirely on ice and quickest lysis, processing and storage. Even though, this good antibody is still highly recommended. Cells from MCF-7 grown under normoxia or hypoxia were lysed for HIF-1 alpha blotting. For the best signal, we suggest higher amount of protein (80-120ug) and traditional tank (wet) electroblotting protocol. 10v overnight transferring is the longest but most efficient way to transfer big proteins as Hif-1a. 1:500 dilution of this Ab and 1:2000 secondary Ab work well.
 
reviewed by:
Anonymous
WB Rat 09/01/2010
View

Summary

ApplicationWestern Blot
Sample Testedendothelial cell nuclear extract, Sample Amount: 40ug
SpeciesRat
CommentsGood results using this Ab for Western blots and ChIP analysis.

Blocking

Blocking DetailsBlocking Buffer: PBS + 5% NFDM + 0.05% Tween 20, Blocking Time: 1 hour, Blocking Temp: room temperature

Primary Anitbody

Dilution RatioDilution Ratio: 1:250, Incubation Dilution Buffer: PBS + 5% NFDM + 0.05% Tween 20, Incubation Time/Temp: overnight at 4C

Secondary Antibody

Secondary DescriptionSecondary Ab: GaM=HRP, Secondary Ab Dilution Ratio: 1:7500

Details

Detection NotesMethod: Chemiluminescence, Exposure Time: 15 min, Positive Control: nuclear extract from same cells, Specific Bands: 60, 98 kDa

Comments

CommentsGood results using this Ab for Western blots and ChIP analysis.
 
reviewed by:
William Lin
WB Mouse 05/17/2010
View

Summary

ApplicationWestern Blot
Sample Testedmouse cortical neurons, Sample Amount: 40ug
SpeciesMouse
LotAA1
CommentsI tested this Hif1a antibody on mouse primary cortical neuronal culture treated with 100uM DFO for over 4 hours. I collected the total lysate using RIPA buffer with 0.1% SDS, sonicated the lysates, and heated up to 90ºC for 5 minutes in SDS-containing sample buffer before loading on an SDS-PAGE gel. I used 1:1000 dilution for the Hif1a antibody (overnight), and 1:20000 anti-mouse secondary antibody for 1 hour. With a few background bands, this antibody works well to detect the upregulation of Hif1a under hypoxic conditions.

Blocking

Blocking DetailsBlocking Buffer: 5% milk in TBST, Blocking Time: 2 hours, Blocking Temp: Room temperature

Primary Anitbody

Dilution RatioDilution Ratio: 1:1000, Incubation Dilution Buffer: blocking buffer, Incubation Time: overnight, Incubation Temp: 4 C

Secondary Antibody

Secondary DescriptionSecondary Ab: anti-mouse IgG-HRP, Secondary Ab Dilution Ratio: 1:20000
Secondary Manufacturer Cat#Promega W402B

Details

Detection NotesDetection Method: ECL, Exposure Time: 2 min, Neg.Control: untreated neurons, Specific Bands: 120 kDa, Non-Specific Bands: 67 kDa

Comments

CommentsI tested this Hif1a antibody on mouse primary cortical neuronal culture treated with 100uM DFO for over 4 hours. I collected the total lysate using RIPA buffer with 0.1% SDS, sonicated the lysates, and heated up to 90ºC for 5 minutes in SDS-containing sample buffer before loading on an SDS-PAGE gel. I used 1:1000 dilution for the Hif1a antibody (overnight), and 1:20000 anti-mouse secondary antibody for 1 hour. With a few background bands, this antibody works well to detect the upregulation of Hif1a under hypoxic conditions.
 
reviewed by:
Anonymous
ICC Other 02/10/2010
View

Summary

ApplicationImmunocytochemistry
Sample TestedHUVEC & Candida albicans
SpeciesOther
LotZ5
CommentsWorks fine with HUVEC but not when Candida albicans is present. The Candida hyphae bind the Ab w/ much greater affinity than the EC and act as a sink leaving the EC completely unstained.

Blocking

Blocking DetailsBlocking Buffer: PBST +1% BSA, Blocking Time: 30 minutes, Blocking Temp: 25 degrees Celsius

Primary Anitbody

Dilution RatioIncubation Dilution Buffer: PBST + 1% BSA, Dilution Ratio: 1/100, Incubation Time: 18 hours, Incubation Temp: 4 degrees Celsius

Secondary Antibody

Secondary DescriptionSecondary Ab: anti-mouse IgG, Secondary Ab Dilution: 1/1000
Secondary Manufacturer Cat#Invitrogen A11004

Comments

CommentsWorks fine with HUVEC but not when Candida albicans is present. The Candida hyphae bind the Ab w/ much greater affinity than the EC and act as a sink leaving the EC completely unstained.

Product General Protocols

Video Protocols

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

FAQs for HIF-1 alpha Antibody (NB100-105). (Showing 1 - 10 of 18 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, as desired by you, I am suggesting you four HIF-1 alpha antibodies based upon customer reviews, as well as the number of peer reviewed publications in which these products has 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 it exist path for detection HIF 1 in the 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. Do you have any suggested IHC Protocol for this antibody? We had been trying to validate protocols from other reserach studies, but failed to get good results. Background staining seems to be the problem.
    • Yes, we have a specific IHC protocol for NB100-105. It is located on the datasheet under the "Protocols & FAQs" tab. We also have over 300 publications that utilize this product. If you need help finding a specific paper or if you would like us to help troubleshoot your experiment, please contact Technical Support at technical@novusbio.com.
  6. Has anyone used this antibody for supershft in EMSA?
    • Yes, this antibody is useful for EMSA and has been documented in couple of recent research publications. You may refer to the following paper for details: Kong D, et al. Echinomycin, a small-molecule inhibitor of hypoxia-inducible factor-1 DNA-binding activity. Cancer Res. 2005 Oct 1;65(19):9047-55 [PMID: 16204079].
  7. What is the difference in the following two HIF-1 alpha antibodies? Catalog # NB100-105 and NB100-134
    • NB100-105 is a mouse monoclonal antibody, whereas NB100-134 is a rabbit polyclonal. They both use the same immunogen.
  8. I am working on a WB using the antibody against mouse HIF1a and HIF2a (catalog numbers NB100-105 and NB100-122). I tried several anti-mouse and anti-rabbit HRP secondary antibodies (from another company) and found they gave different patterns. Please provide the secondary antibody info which was used in the WB showed on the datasheet.
    • We use catalog numbers NB7570 and NB7183 as our secondary antibodies for testing these antibodies in WB.
  9. What is the difference between NB100-105 and NB100-123?
    • NB100-105 and NB100-123 are similar as far as immunogens and clones are concerned, however, they are different with respect to their purification process, concentrations and recommended dilutions. NB100-123 is Protein A purified IgG2b with a concentration of 1.5 mg/ml, whereas NB100-105 is Protein G purified IgG2b with 4.0 mg/ml concentration. Another difference is the recommended dilutions (because of the differences in their concentration).
  10. Can I get a sample of the HIF1A antibody (NB100-105)?
    • We do offer this antibody is a Sample Size, which contains 0.025ml of antibody. The sample size catalog number is NB100-105SS.
  11. Show All 18 FAQs.

Positive Control Lysate(s)

Secondary Antibodies

 

Isotype Controls

Other Available Formats

Alexa Fluor 405 Labeled NB100-105AF405
Alexa Fluor 488 Labeled NB100-105AF488
Alexa Fluor 647 Labeled NB100-105AF647
Alexa Fluor 700 Labeled NB100-105AF700
Biotin Labeled NB100-105B
DyLight 405 Labeled NB100-105V
DyLight 488 Labeled NB100-105G
DyLight 550 Labeled NB100-105R
DyLight 650 Labeled NB100-105C
FITC Labeled NB100-105F
HRP Labeled NB100-105H

Additional HIF-1 alpha Antibody (H1alpha67) Products

Related Products by Gene

Bioinformatics Tool for HIF-1 alpha Antibody (NB100-105)

Discover related pathways, diseases and genes to HIF-1 alpha Antibody (NB100-105). Need help? Read the Bioinformatics Tool Guide for instructions on using this tool.
Visit Tool

Diseases for HIF-1 alpha Antibody (NB100-105)

Discover more about diseases related to HIF-1 alpha Antibody (NB100-105).
 

Pathways for HIF-1 alpha Antibody (NB100-105)

View related products by pathway.

PTMs for HIF-1 alpha Antibody (NB100-105)

Learn more about PTMs related to HIF-1 alpha Antibody (NB100-105).
 

Research Areas for HIF-1 alpha Antibody (NB100-105)

Find related products by research area.

Blogs on HIF-1 alpha. Showing 1-10 of 25 blog posts - Show all blog posts.

The relationship between Ki67 and HIF-1 in cancer
Ki67, also known as MKI67, is best known as the leading marker of cellular proliferation. Ki67 is regulated by a balance between synthesis and degradation, and often carries a very short half-life.  First discovered to be located to dividing cells,...  Read full blog post.

Controls for HIF-1 Alpha's WB, ICC-IF, IHC, IP & FLOW Analysis
Tips on positive and negative controls for HIF-1 alpha antibodies is one of the most Frequently Asked Questions on Hypoxia and HIFs. Here are top 5 suggestions from Novus Biologicals: The degradation of HIF1 alpha is the most common issue whic...  Read full blog post.

Understanding the relationship between HIF-1 alpha, Hypoxia and Epithelial-Mesenchymal Transition
Epithelial-mesenchymal transition (EMT) is a natural process by which epithelial cells lose their polarity and intercellular adhesion, and gain the migratory invasive properties of mesenchymal stem cells that can differentiate into a variety of cel...  Read full blog post.

HIF-2 alpha: HIF1A's Homologue with Similar and Divergent Functions
HIF-2 alpha is a member of the heterodimeric hypoxia-inducible factors/HIFs family (HIF-1, HIF-2, and HIF-3) which contains a common beta subunit but differ in their alpha subunits. Also called as EPAS1 or Mop2, HIF-2 alpha regulates cellular adapt...  Read full blog post.

HIF-3 alpha: a versatile target with hypoxia dependent and independent functions
By: Subhash GangarHIF-3 alpha (hypoxia-inducible factor 3-alpha/ HIF3A) represents an isoform of HIF-alpha subunits which heterodimerize with stable beta subunit (HIF-beta) for the regulation of HIF target genes through binding to hypoxia respon...  Read full blog post.

HIF-1 alpha - sensing and responding to changing oxygen levels
Hypoxia-inducible factor 1 (HIF-1) allows cells to respond to changing levels of oxygen in the environment. HIF-1 is a heterodimeric transcription factor consisting of alpha and beta subunits. Under normal conditions HIF-1 alpha is continuously sy...  Read full blog post.

Comprehensive Autophagy Research Tools - New Catalog Available Now!
Autophagy, a protein degradation process through autophagosome-lysosomal pathway, is important for cellular homeostasis and plays a role in many diseases. To help researchers learn more about this process and the products available for its study, N...  Read full blog post.

HIF-1 beta: A dimerization partner of HIF-1 alpha required for an adaptive response to hypoxia
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. Hypoxia-indu...  Read full blog post.

HIF-1 Alpha: Infographic
Encoded by the HIF1A gene, HIF-1 alpha has a critical role in cellular response to hypoxia. In hypoxic conditions, HIF-1 alpha activates the transcription of several genes to facilitate metabolic reaction for lack of oxygen. In normoxic conditions, HI...  Read full blog post.

Controlling the HIF-1 Switch
Hypoxia-inducible factor-1 is a major transcription factor composed of two subunits: HIF-1alpha and HIF-1 beta. Under normoxic conditions, HIF-1 alpha is targeted to proteosomal degradation via ubiquitination. On the other hand during hypoxic conditio...  Read full blog post.

Showing 1-10 of 25 blog posts - Show all blog posts.

Contact Information

Product PDFs

Bioinformatics

Gene Symbol HIF1A
Entrez
OMIM

Customer Resources

Novus Review - Submit your review and earn rewards points which can be used for merchandise & discounts.
Risk Free Testing - Test on a species/application not listed above to receive a full credit towards a future purchase.

Novus' Quality Guarantee - Novus guarantees that every product we sell will work in the application and species listed on our website and datasheets.

Submit your question on NB100-105 below.
During business hours, we will respond to your email within 24 hours. For any questions submitted on the weekend, a response will be received on Monday.
For immediate assistance during business hours M- F (excluding major holidays), please contact us.
Ask a Scientist - We have a lab full of white coats just waiting for your scientific questions and concerns.

Customers Who Bought This Also Bought