HIF-1 alpha Antibody Pack

Western Blot: HIF-1 alpha Antibody Pack [NB100-901IHC] - Detection of mouse HIF1-alpha on hypoxia treated MEFs using NB 100-449.

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody Pack [NB100-901IHC] - Detection of HIF-1 alpha (red dye 568) in a cultured raw mouse macrophage cell line, using NB100-131. 100X magnification. Photos courtesy of Susan Alexander and Hattie Gresham, PhD.

Immunohistochemistry: HIF-1 alpha Antibody Pack [NB100-901IHC] - Staining of placenta, villi using.

Flow Cytometry: HIF-1 alpha Antibody Pack [NB100-901IHC] - Analysis of HIF1-alpha in CoCl2 Treated vs. Untreated HeLa Cells. Hela cells were treated for 15 hrs with 200uM CoCl2, fixed in PFA, and permeabilized in 90% MeOH. One million cells were stained with 0.125ug of NB100-449 and secondary FITC-conjugated goat anti-rabbit (in a 150ul reaction). Black- treated, anti-KLH control IgG; Reduntreated, anti-HIF1-alpha; Blue- treated, anti-HIF1-alpha.

Western Blot: HIF-1 alpha Antibody Pack [NB100-901IHC] - Detection of HIF-1 alpha in cobalt chloride treated/untreated COS-7 nuclear extracts.

Western Blot: HIF-1 alpha Antibody Pack [NB100-901IHC] - On day 1, MEF cells (+/+,-/-), were grown on 15cm dish (2x10 to the 6th cells). On day 2, cells were exposed to hypoxia for 4hrs. Cells were washed with ice cold PBS twice and whole cell protein was extracted with RIPA buffer fortified with protease. Upon quantification, 100ug of protein was fractionated on 7% polyacralymide gel. -Gel was transferred overnight onto nitrocellulose membrane. The membrane was probed with HIF-1 alpha monoclonal antibody at a 1:500 dilution (NB 100-123). The secondary antibody was conjugated with HRP and was used at a 1:2500 dilution. Photo courtesy of Dr. Gregg Semenza, Johns Hopkins University.

Western Blot: HIF-1 alpha Antibody Pack [NB100-901IHC] - 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) NB100-449.

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody Pack [NB100-901IHC] - Detection of HIF-1 alpha (red dye) in a cell cytospin from a lavage of a murine skin pouch infected with Staph Aureus, using NB100-131. Blue dye is DAPI nuclear staining. Photos courtesy of Susan Alexander and Hattie Gresham, PhD.

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody Pack [NB100-901IHC] - Detection of HIF-1 alpha (red dye) in a cell cytospin from a lavage of a murine skin pouch infected with Staph Aureus, using NB100-131. 100X magnification. Blue dye is DAPI nuclear staining. Photos courtesy of Susan Alexander and Hattie Gresham, PhD.

Immunocytochemistry/Immunofluorescence: HIF-1 alpha Antibody Pack [NB100-901IHC] - Detection of HIF-1 alpha (red dye 568) in a cultured raw mouse macrophage cell line. Photos courtesy of Susan Alexander and Hattie Gresham, PhD.

Immunohistochemistry: HIF-1 alpha Antibody Pack [NB100-901IHC] - Staining of HIF-1 alpha in human kidney.

Immunohistochemistry: HIF-1 alpha Antibody Pack [NB100-901IHC] - Staining in hypoxia-induced human placenta.

Immunohistochemistry-Paraffin: HIF-1 alpha Antibody Pack [NB100-901IHC] - Negative control stain of human placenta (from sea level) using mouse IgG at 1:100. 4uM paraffin-embedded section.

Immunohistochemistry: HIF-1 alpha Antibody Pack [NB100-901IHC] - Staining of mouse brain, neurons using NB100-449.

Immunoprecipitation: HIF-1 alpha Antibody Pack [NB100-901IHC] - Immunoprecipitation of HIF-1alpha using NB100-123. Heavy and light chains are also detected.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Flow, ICC/IF, IHC, IHC-Fr, IHC-P, IP

HIF-1 alpha Antibody Pack Summary

• Immunogen: NB 100-131: Human HIF-1 alpha amino acids 329-530. NB100-123: Fusion protein containing amino acids 432-528 of human HIF-1alpha. NB100-449: peptide between residues 775 and the C-terminus (residue 826) of human HIF-1 alpha NB100-479: A fusion protein
• Specificity: These antibodies are specific for HIF-1alpha.
• Gene: HIF1A

Applications/Dilutions

• Application Notes: Immunohistochemical staining of paraffin-embedded and frozen sections (only NB 100-131 has been used on frozen sections). Antigen retrieval is strongly recommended. With the exception of NB100-449 and NB100-479 (which are also recommended for Western Blot), these antibodies work for Western blot analysis but are not specifically recommended for this application. A starting dilution of 1:1,000 is recommended with titration up or down depending upon the signal and background.

Packaging, Storage & Formulations

• Storage: Store at 4C. Do not freeze.

Notes

1 sample size vial each of: NB100-479 (0.025mL), NB100-449 (0.025mL), NB100-123 (0.05mL) and NB100-131 (0.05mL).

Alternate Names for HIF-1 alpha Antibody Pack

• 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
• HIF1A
• HIF-1a
• 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

Our HIF-1 alpha sample pack provides a convenient way to determine the optimal HIF-1 alpha antibody for your specific species and tissues. Hypoxia contributes significantly to the pathophysiology of major categories of human disease, including myocardial and cerebral ischemia, cancer, pulmonary hypertension, congenital heart disease and chronic obstructive pulmonary disease. HIF-1 is a nuclear protein that activates gene transcription in response to reduced cellular 02 concentration. HIF-1 activates the transcription of EPO, VEGF, iNOS, heme oxygenase 1 and several other critical intracellular responses to hypoxia. HIF-1 is a heterodimer composed of HIF-1 alpha and HIF-1 beta subunits. Both subunits are induced by hypoxia and rapidly decay upon return to normoxia and are basic-helix-loop-helix-PAS proteins. Recent research indicates the ability to regulate hypoxia-inducible factors may be related to tumor-related angiogenesis in certain cancers.

Limitations

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

Publications for HIF-1 alpha Antibody Pack (NB100-901IHC)(5)

Publications using NB100-901IHC

• Hill, P et al. Inhibition of hypoxia inducible factor hydroxylases protects against renal ischemia-reperfusion injury. J Am Soc Nephrol;19(1):39-46. 2008 Jan. [PMID: 18178798]
• Higami, Y et al. Energy restriction lowers the expression of genes linked to inflammation, the cytoskeleton, the extracellular matrix, and angiogenesis in mouse adipose tissue. J Nutr;136(2):343-52. 2006 Feb. [PMID: 16424110]
• Tanaka, T et al. Hypoxia-inducible factor modulates tubular cell survival in cisplatin nephrotoxicity. Am J Physiol Renal Physiol289(5):F1123-33. 2005 Nov. [PMID: 15956779]
• Lee, S, Kim, SJ, Kim, DJ et al. The Clinical Significanse of the Hypoxia Inducible Factor-1 C4d in Implantation Renal Biopsies Post-Transplant Biopsies. 22nd International Congress of the Transplantation Society. 10-Aug-08.
• Quintero, M et al. Nitric oxide is a factor in the stabilization of hypoxia-inducible factor-1alpha in cancer: role of free radical formation. Cancer Res;66(2):770-4. 2006 Jan 15. [PMID: 16424008]

FAQs for HIF-1 alpha Antibody Pack (NB100-901IHC). (Showing 1 - 5 of 5 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. We have received an enquiry regarding HIF1 alpha antibody panel, product code NB 100-901IHC. I would appreciate if you are able to provide the WB testing protocol for this antibody.
   • There are four antibodies in this pack. We have specific western blot protocols for 3 of the 4 antibodies, as follows: Western Blot protocol specific for NB100-123, Western Blot protocol specific for NB100-479, Western Blot protocol specific for NB100-449. You may find our general western blot protocol useful.

Bioinformatics

• Gene Symbol: HIF1A
• Entrez:
  • 3091(Human)
  • 15251(Mouse)
  • 29560(Rat)