HIF-1 alpha [p Ser311, Hydroxy Thr299] Antibody (Clone) [mFluor Violet 450 SE] - (Asp175), (CoCl2 Treated), K75D Mutant - Azide and BSA Free

Western Blot: HIF-1 alpha [p Ser311, Hydroxy Thr299] Antibody (Clone) [Alexa Fluor® 555] - (Asp175) (CoCl2 Treated) - Pre-adsorbed [NB100-105L] - test1234

Immunohistochemistry: HIF-1 alpha [p Ser311, Hydroxy Thr299] Antibody (H1) [Alexa Fluor® 488] - (Asp175), (CoCl2 Treated) - Pre-adsorbed [NB100-105L] - test

Flow Cytometry: HIF-1 alpha [p Ser311, Hydroxy Thr299] Antibody (H1) [Alexa Fluor® 488] - (Asp175), (CoCl2 Treated) - Pre-adsorbed [NB100-105L] - test

Western Blot: HIF-1 alpha [p Ser311, Hydroxy Thr299] Antibody (H1) [Alexa Fluor® 488] - (Asp175), (CoCl2 Treated) - Pre-adsorbed [NB100-105L] - test

Western Blot: HIF-1 alpha [p Ser311, Hydroxy Thr299] Antibody (H1) [Alexa Fluor® 488] - (Asp175) (CoCl2 Treated) - Pre-adsorbed [NB100-105L] - Western Blot

Western Blot: HIF-1 alpha [p Ser311, Hydroxy Thr299] Antibody (H1H1) [Alexa Fluor® 488] - (Asp175), (CoCl2 Treated) - Pre-adsorbed [NB100-105L] - Test

Western Blot: HIF-1 alpha [p Ser311, Hydroxy Thr299] Antibody (H1H1) [Alexa Fluor® 488] - (Asp175), (CoCl2 Treated) - Pre-adsorbed [NB100-105L] - Test

Testing123

Product Discontinued

HIF-1 alpha [p Ser311, Hydroxy Thr299] Antibody (Clone) [mFluor Violet 450 SE] - (Asp175), (CoCl2 Treated), K75D Mutant - Azide and BSA Free Summary

• Description: DRAQ7 (TM) is an ideal far-red DNA counterstain for IF/IHC, high content screening and cell-based assays. DRAQ7 (TM) has many applications and is highly compatible with existing protocols across a wide range of instrumentation platforms.
  
  Key Features of DRAQ7 (TM):
  
  Does NOT enter intact, live cells
  Excited by blue through red lasers
  DRAQ7 (TM) is NON-TOXIC in long-term culture
  Ideal replacement for propidium iodide (and 7-AAD) having far better spectral properties: no UV excitation and no emission overlap with PE and homologues
  Allows a new design of combination analysis for cell health and status
  Viability/integrity can be directly interchanged with DRAQ5 (TM) (DNA content)
  Rapid staining of dsDNA/nuclei of DEAD or permeabilised cells
  Combination with live cell dyes for dead/live discrimination
  Easy to use - no wash, no RNase needed
  Ideal for use with GFP & FITC labels - DRAQ7 (TM) fluoresces in the far-red region
  NO fluorescence enhancement upon DNA binding
  Low photobleaching effect
  Compatibility with optics of flow, laser scanning cytometers and confocal and lamp-based fluorescence microscopes
  Cost-effective analysis
• Background: BG2 - test
• Immunogen: Immunogen
• Epitope: EpitopeEpitope
• Modification: p Ser311, Hydroxy Thr299
• Target Species: Human
• Localization: Localization
• Marker: Marker
• Specificity: SpecificitySpecificitySpecificity
• Preparation Method: Preparation Method
• Details of Functionality: Details of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails of FunctionalityDetails
• Source: Bovine Brain
• Predicted Species: Baboon (98%), Algae (95%), Armenian Hamster (100%). Backed by our 100% Guarantee.
• Selection Agent: Selection Agent
• Growth Properties: Growth Properties
• Content: Inhibitor Content
• Standard Curve Range: Standard Curve Range
• Sensitivity: Sensitivity
• Assay Type: Assay Type
• Inter-Assay: Inter Assay
• Intra-Assay: Intra Assay
• Spike Recovery: Spiking Recovery
• Sample Volume: Sample Volume
• Kit Type: Antibody Labeling Kit
• Protein/Peptide Type: Enzyme Protein Fractions
• RCL Type: Stable Cell Line
• Isotype: IgG2b
• Clonality: Polyclonal
• Host: Pig
• Gene: HIF1A
• Purity: Unpurified
• Endotoxin Note: Endotoxin

Applications/Dilutions

• Dilutions:
  • Binding Activity
  • Cell Culture 1:1000 - 1:20000
  • CyTOF-ready 1x10^6
  • Dual RNAscope ISH-IHC Compatible
  • ELISA 1:100 - 1:2000
  • Immunocytochemistry/ Immunofluorescence 1:100
  • Immunohistochemistry-Frozen 1:20 - 1:50
  • Immunohistochemistry-Paraffin 100 ug/ml for 30 minutes at RT
  • Western Blot 1:500
• Application Notes: Application Notes
• Theoretical MW: 20/70 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.
• Growth Media:
  • NB500-201AF405 NB500-201AF405
• Agonist:
  • Survivin Antibody - BSA Free NB500-201
• Antagonist:
  • ATM Antibody - BSA Free NB100-104
  • KLF8 Antibody NB100-1050
• Readout System:
  • HIF-1 alpha Antibody (H1alpha67) [7C] NB100-105V2
• Control:

  HIF-1 alpha Antibody (H1alpha67)
  HIF-1 alpha Antibody (H1alpha67)
  COS-7 Nuclear Hypoxic Induced Cell Lysate

Reactivity Notes

Species Reactivity

Packaging, Storage & Formulations

• Storage: Store at 4C in the dark.
• Buffer: BufferBuffer - making a test edit
• Preservative: 0.05% Sodium Azide
• Concentration: This product is unpurified. The exact concentration of antibody is not quantifiable.
• Purity: Unpurified
• Reconstitution Instructions: Reconstitution instructions.Reconstitution instructions.Reconstitution instructions.

Details for Array

• Type: Knockout 293T Cell
• Life Stage: Post-Natal
• Tissue: Blood Lymph
• Protein State: Poorly Differentiated
• Tissue Condition: Astrocytoma Grade I
• Subcellular Fraction: DTT Treated / Untreated
• Position: Ventral

Notes

Public Notes This product is provided under an intellectual property license from Life Technologies Corporation. The purchase of this product conveys to the buyer the non-transferable right to use the purchased product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components, or any materials made using the product or its components, in any activity to generate revenue, which may include, but is not limited to use of the product or its components: (i) in manufacturing; (ii) to provide a service, information, or data in return for payment; (iii) for therapeutic, diagnostic or prophylactic purposes; or (iv) for resale, regardless of whether they are resold for use in research. For information on purchasing a license to this product for purposes other than as described above, contact Life Technologies Corporation, 5791 Van Allen Way, Carlsbad, CA 92008 USA or outlicensing@lifetech.com.

Alternate Names for Array

• 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
• H1alpha67
• 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

Test123

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.

Publications for HIF-1 alpha Antibody (NB100-105L)(2)

Publications using NB100-105L

• Florescu C, Tartea G, Streba C Et Al. The Evaluation of Beta-2-Adrenoreceptors' Expression in Normal Peritumoral Tissue in Patients with Colorectal Adenocarcinoma. Curr Health Sci J 2018-12-25 [PMID: 30581586]
• Yang TH, Yao HT, Chiang MT. Red algae (Gelidium amansii) hot-water extract ameliorates lipid metabolism in hamsters fed a high-fat diet. J Food Drug Anal 2017-10-09 [PMID: 28987370]

Review for HIF-1 alpha Antibody (NB100-105L) (1)

Average Rating: 5

(Based on 1 review)

We have 1 review tested in 1 species: Mouse.

reviewed by: Verified Customer WB Mouse 03/04/2014

Summary

• Application: Western Blot
• Sample Tested: samples tested
• Species: Mouse
• Lot: 123456

FAQs for HIF-1 alpha Antibody (NB100-105L). (Showing 1 - 10 of 11 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. Is cross-reactivity with HIF-2 alpha tested/predicted?
    • Although we don’t have cross-reactivity data with regards to HIF-2 alpha, we predict minimal cross-reactivity based on low sequence similarity observed from BLAST analysis between HIF-1 alpha and HIF-2 alpha.
Show All 11 FAQs.

Bioinformatics

• Gene Symbol: HIF1A
• Entrez:
  • 11560(Avian)
  • 11100008(Bovine)
• OMIM:
  • 5678(Human)
• Uniprot:
  • P63244()
  • Q02223()
  • Q16665()
  • Q16665()
  • Q9XTW2(C. elegans)
  • P31944(Human)
  • O97703(Monkey)