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Cardiovascular Hypoxia HIF
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Anti-HIF-1 alpha antibody [ESEE122] (ab8366)

  • Datasheet
  • SDS
Reviews (2)Q&A (8)References (40)

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF-1 alpha antibody [ESEE122] (ab8366)
  • Immunocytochemistry/ Immunofluorescence - Anti-HIF-1 alpha antibody [ESEE122] (ab8366)

Key features and details

  • Mouse monoclonal [ESEE122] to HIF-1 alpha
  • Suitable for: ICC/IF, IHC-P
  • Reacts with: Human
  • Isotype: IgG1

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Overview

  • Product name

    Anti-HIF-1 alpha antibody [ESEE122]
    See all HIF-1 alpha primary antibodies
  • Description

    Mouse monoclonal [ESEE122] to HIF-1 alpha
  • Host species

    Mouse
  • Tested Applications & Species

    Application Species
    ICC/IF
    Human
    IHC-P
    Human
    See all applications and species data
  • Immunogen

    Recombinant fragment corresponding to Human HIF-1 alpha aa 300-550.

  • Positive control

    • IHC-P: Human colon tissue. ICC: HeLa cells.
  • General notes

    This antibody clone is manufactured by Abcam.

    Under normoxic conditions HIF-1 alpha has a short half-life. It is largely undetectable in cells or tissues grown under normoxic conditions. It is stabilized only at O2 concentrations below 5% and upon stabilization under hypoxic conditions HIF-1 translocates to the nucleus. Therefore we recommend western blots using nuclear extracts and running Hypoxia treated samples as positive control (ab180880). Hypoxia can be induced with treatment using certain agents e.g. CoCl2 or DFO, etc. so proper sample preparation is critical.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • Purity

    Protein G purified
  • Clonality

    Monoclonal
  • Clone number

    ESEE122
  • Myeloma

    NS1
  • Isotype

    IgG1
  • Light chain type

    unknown
  • Research areas

    • Cardiovascular
    • Hypoxia
    • HIF
    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • HLH / Leucine Zipper
    • HLH
    • Epigenetics and Nuclear Signaling
    • Cardiovascular/Immune
    • Hypoxia
    • HIF
    • Cancer
    • Invasion/microenvironment
    • Hypoxia
    • HIF
    • Cancer
    • Cancer Metabolism
    • Response to hypoxia
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Nucleotide metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Hypoxia
    • Metabolism
    • Types of disease
    • Cancer

Associated products

  • Alternative Versions

    • HRP Anti-HIF-1 alpha antibody [ESEE122] (ab215773)
  • Assay kits

    • HIF-1 alpha Transcription Factor Assay Kit (ab133104)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat Anti-Mouse IgG H&L (DyLight® 488) preadsorbed (ab96879)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
    • Mouse IgG1, Kappa Monoclonal [B11/6] - Isotype Control (ab91353)
  • Recombinant Protein

    • Recombinant Human HIF-1 alpha protein (ab154478)
    • Recombinant Human HIF-1 alpha protein (ab48734)
  • Related Products

    • HeLa-Vehicle treated (Negative Control) Whole Cell Lysate (ab116321)
    • Human HIF-1 alpha ELISA Kit (ab171577)
    • HIF-1 alpha (H1alpha67, EP1215Y, ESEE122, EPR16897) Antibody Panel (ab263460)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab8366 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC/IF
Human
IHC-P
Human
Application Abreviews Notes
ICC/IF
Use a concentration of 8 - 12 µg/ml.
IHC-P (2)
1/1000 - 1/8000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Notes
ICC/IF
Use a concentration of 8 - 12 µg/ml.
IHC-P
1/1000 - 1/8000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for WB.
  • Target

    • Function

      Functions as a master transcriptional regulator of the adaptive response to hypoxia. Under hypoxic conditions activates the transcription of over 40 genes, including, erythropoietin, glucose transporters, glycolytic enzymes, vascular endothelial growth factor, and other genes whose protein products increase oxygen delivery or facilitate metabolic adaptation to hypoxia. Plays an essential role in embryonic vascularization, tumor angiogenesis and pathophysiology of ischemic disease. Binds to core DNA sequence 5'-[AG]CGTG-3' within the hypoxia response element (HRE) of target gene promoters. Activation requires recruitment of transcriptional coactivators such as CREBPB and EP300. Activity is enhanced by interaction with both, NCOA1 or NCOA2. Interaction with redox regulatory protein APEX seems to activate CTAD and potentiates activation by NCOA1 and CREBBP.
    • Tissue specificity

      Expressed in most tissues with highest levels in kidney and heart. Overexpressed in the majority of common human cancers and their metastases, due to the presence of intratumoral hypoxia and as a result of mutations in genes encoding oncoproteins and tumor suppressors.
    • Sequence similarities

      Contains 1 basic helix-loop-helix (bHLH) domain.
      Contains 1 PAC (PAS-associated C-terminal) domain.
      Contains 2 PAS (PER-ARNT-SIM) domains.
    • Domain

      Contains two independent C-terminal transactivation domains, NTAD and CTAD, which function synergistically. Their transcriptional activity is repressed by an intervening inhibitory domain (ID).
    • Post-translational
      modifications

      In normoxia, is hydroxylated on Pro-402 and Pro-564 in the oxygen-dependent degradation domain (ODD) by EGLN1/PHD1 and EGLN2/PHD2. EGLN3/PHD3 has also been shown to hydroxylate Pro-564. The hydroxylated prolines promote interaction with VHL, initiating rapid ubiquitination and subsequent proteasomal degradation. Deubiquitinated by USP20. Under hypoxia, proline hydroxylation is impaired and ubiquitination is attenuated, resulting in stabilization.
      In normoxia, is hydroxylated on Asn-803 by HIF1AN, thus abrogating interaction with CREBBP and EP300 and preventing transcriptional activation. This hydroxylation is inhibited by the Cu/Zn-chelator, Clioquinol.
      S-nitrosylation of Cys-800 may be responsible for increased recruitment of p300 coactivator necessary for transcriptional activity of HIF-1 complex.
      Requires phosphorylation for DNA-binding.
      Sumoylated; by SUMO1 under hypoxia. Sumoylation is enhanced through interaction with RWDD3. Desumoylation by SENP1 leads to increased HIF1A stability and transriptional activity.
      Ubiquitinated; in normoxia, following hydroxylation and interaction with VHL. Lys-532 appears to be the principal site of ubiquitination. Clioquinol, the Cu/Zn-chelator, inhibits ubiquitination through preventing hydroxylation at Asn-803.
      The iron and 2-oxoglutarate dependent 3-hydroxylation of asparagine is (S) stereospecific within HIF CTAD domains.
    • Cellular localization

      Cytoplasm. Nucleus. Cytoplasmic in normoxia, nuclear translocation in response to hypoxia. Colocalizes with SUMO1 in the nucleus, under hypoxia.
    • Target information above from: UniProt accession Q16665 The UniProt Consortium
      The Universal Protein Resource (UniProt) in 2010
      Nucleic Acids Res. 38:D142-D148 (2010) .

      Information by UniProt
    • Database links

      • Entrez Gene: 3091 Human
      • Omim: 603348 Human
      • SwissProt: Q16665 Human
      • Unigene: 597216 Human
      • Alternative names

        • ARNT interacting protein antibody
        • ARNT-interacting protein antibody
        • Basic helix loop helix PAS protein MOP1 antibody
        • Basic-helix-loop-helix-PAS protein MOP1 antibody
        • bHLHe78 antibody
        • Class E basic helix-loop-helix protein 78 antibody
        • HIF 1A antibody
        • HIF 1alpha antibody
        • HIF-1-alpha antibody
        • HIF-1alpha antibody
        • HIF-alpha antibody
        • HIF1 A antibody
        • HIF1 Alpha antibody
        • HIF1 antibody
        • HIF1-alpha antibody
        • HIF1A antibody
        • HIF1A_HUMAN antibody
        • hifla antibody
        • Hypoxia inducible factor 1 alpha antibody
        • Hypoxia inducible factor 1 alpha isoform I.3 antibody
        • Hypoxia inducible factor 1 alpha subunit antibody
        • Hypoxia inducible factor 1 alpha subunit basic helix loop helix transcription factor antibody
        • Hypoxia inducible factor 1, alpha subunit (basic helix loop helix transcription factor) antibody
        • Hypoxia inducible factor1alpha antibody
        • Hypoxia-inducible factor 1-alpha antibody
        • Member of PAS protein 1 antibody
        • Member of PAS superfamily 1 antibody
        • Member of the PAS Superfamily 1 antibody
        • MOP 1 antibody
        • MOP1 antibody
        • PAS domain-containing protein 8 antibody
        • PASD 8 antibody
        • PASD8 antibody
        see all

      Images

      • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF-1 alpha antibody [ESEE122] (ab8366)
        Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF-1 alpha antibody [ESEE122] (ab8366)

        IHC image of HIF-1-alpha staining in human normal colon formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab8366, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.

         

        For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

        *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

      • Immunocytochemistry/ Immunofluorescence - Anti-HIF-1 alpha antibody [ESEE122] (ab8366)
        Immunocytochemistry/ Immunofluorescence - Anti-HIF-1 alpha antibody [ESEE122] (ab8366)

        ab8366 stained Hela cells. The cells were 4% formaldehyde fixed for 10 minutes, permeabilized in 0.1% PBS-Triton X-100 for 5 min and then blocked in 1%BSA / 10% normal Goat serum / 0.3M glycine in 0.1% PBS-Tween for 1 hour at room temperature to block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab8366 at 10µg/ml) overnight at +4°C. The secondary antibody (pseudo-colored green) was ab150117 Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed  used at a 1/1000 dilution for 1hour at room temperature. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1hour at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

      Protocols

      • Recommended IHC-P protocol with ab8366
      • Mouse on Mouse staining protocol

      Click here to view the general protocols

      Datasheets and documents

      • SDS download

      • Datasheet download

        Download

      References (40)

      Publishing research using ab8366? Please let us know so that we can cite the reference in this datasheet.

      ab8366 has been referenced in 40 publications.

      • Hao S  et al. 2-Methoxyestradiol attenuates chronic-intermittent-hypoxia-induced pulmonary hypertension through regulating microRNA-223. J Cell Physiol 234:6324-6335 (2019). PubMed: 30246291
      • Ribeiro GP  et al. Comparison between two programs for image analysis, machine learning and subsequent classification. Tissue Cell 58:12-16 (2019). PubMed: 31133239
      • Lu L  et al. Inhibitor of growth 4 (ING4) inhibits hypoxia-induced EMT by decreasing HIF-1a and snail in HK2 cells. Acta Histochem 121:695-703 (2019). PubMed: 31239073
      • Han S  et al. The prognostic value of hypoxia-inducible factor-1a in advanced cancer survivors: a meta-analysis with trial sequential analysis. Ther Adv Med Oncol 11:1758835919875851 (2019). PubMed: 31579115
      • La Porta S  et al. Endothelial Tie1-mediated angiogenesis and vascular abnormalization promote tumor progression and metastasis. J Clin Invest 128:834-845 (2018). PubMed: 29355844
      View all Publications for this product

      Customer reviews and Q&As

      Show All Reviews Q&A
      Submit a review Submit a question

      1-10 of 10 Abreviews or Q&A

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-HIF-1-alpha antibody [ESEE122]

      Good
      Abreviews
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
      Blocking step
      Serum as blocking agent for 45 minute(s) · Concentration: 1.5% · Temperature: 25°C
      Antigen retrieval step
      Enzymatic
      Sample
      Rat Tissue sections (Gonad)
      Specification
      Gonad
      Permeabilization
      No
      Fixative
      Formaldehyde
      Read More

      Abcam user community

      Verified customer

      Submitted Mar 13 2014

      Question

      Product code: 8366

      Inquiry: Dear ladies and gentlemen, concerning the positive control to your Antibody against HIF1a (clone ESEE122), what is meant in detail by "hypoxia induced placenta"? Placenta cultivated at lox oxygen concentrations? For how long? Term placenta or first trimester placenta? Which oxygen concentration? Which part of the placenta? Thank you very much in advance for your help!

      Read More

      Abcam community

      Verified customer

      Asked on Aug 10 2012

      Answer

      Thank you for your inquiry.

      The placental tissue was a rather unique sample of which we know of no commercial source of and also did not get any more information form the scientist that used it.

      For an IHC positive control we recommend using glioblastoma multiformae.

      I am sorry I could not be of more help and hope this information is nevertheless helpful.

      Read More

      Abcam Scientific Support

      Answered on Aug 10 2012

      Question

      Is there more information about the IF cytospin protocol for the images shown on the datasheet?

      Read More

      Abcam community

      Verified customer

      Asked on Jun 26 2012

      Answer

      Thank you for your call yesterday and for your patience while I have been in touch with the lab regarding the cytospin staining protocol for the images shown on the datasheet.

      We do not have much more information about the protocol, as the cytospin stains were done by an outside lab in 2005 and we did not receive the full protocol. The primary antibody was conjugated to a fluorophore prior to the staining, so no secondary antibody was used. We do not have information about the how the cytospin was prepared.

      I am sorry that we don't have further information regarding this protocol, but if there is anything else that we might be able to do for you please let me know and I'll be happy to help.

      Read More

      Abcam Scientific Support

      Answered on Jun 26 2012

      Question

      ab is staining the negative tissues also
      Samples- Prostate cancer tissue sections and Normal prostate cancer tissue sections
      IHC-P application
      Microwave antigen retrieval method for 30 minutes.
      Dilution 1/10,000
      The antibody use to work previously

      Read More

      Abcam community

      Verified customer

      Asked on Jan 20 2012

      Answer

      Thank you very much for your phone call last week.

      HiF1 alpha is a very sensitive target it stabilizes under hypoxic conditions and gets degraded under normoxic conditions. Upon stabilization HIF-1 translocates to the nucleus so we recommend permeabilizing the sections.

      I have attached the protocol that we recommend for HIF1 alpha target staining. Although this protocol is not different than others so may be troubleshooting few steps will helps
      - Antigen retrieval always needs optimizations. 20 minutes is only a suggested antigen retrieval time for Microwave. Less than 20 minutes may leave the antigens under retrieved, leading to weak staining. More than 20 minutes may leave them over-retrieved, leading to nonspecific background staining and also increasing the chances of sections dissociating from the slides. A control experiment is recommended beforehand, where slides of the same tissue section are retrieved for 5, 10, 15, 20, 25 and 30 minutes before being immunohistochemically stained to evaluate optimum antigen retrieval time for the particular antibody being used.
      - We would recommend using citrate buffer as antigen retrieval buffer however for optimizations purpose different buffers can be used e.g. EDTA, Tris EDTA.
      - Tissues sections: the tissue should be fixed immediately after removal; any delays actually degrades the protein.
      - We suggest incubating the section in Normal immune serum or 5% BSA for at least 1-2 hours.
      - Following publications may also help with protocol
      http://ajpheart.physiology.org/content/289/5/H2066.long
      http://www.ncbi.nlm.nih.gov/pmc/articles/PMC468666/?tool=pubmed
      Same clone number has been mentioned in following publications
      Beasley et al. 2002. Cancer Res. 62:2493-7. PMID: 11980639
      Giatromanolaki et al. 2003. J. Pathol. 200:222-8. PMID: 12754744
      Hui et al. 2002. Clin Cancer Res. 8:2595-604. PMID: 12171890
      I hope these suggestions will help. Should you have any further question please do not hesitate to contact me.

      Read More

      Abcam Scientific Support

      Answered on Jan 23 2012

      Question

      I need an anti-HIF antibody that will work in IHC-Fr on mouse tissue. I also need positive controls for this.

      Read More

      Abcam community

      Verified customer

      Asked on Dec 22 2011

      Answer

      Thank you for contacting Abcam. I have had a chance to look through the data for at HIF1 alpha antibody panel (ab10625) that you had spoke of during our conversation. Of the antibodies included in this panel only ab8366 (Anti-HIF1 alpha antibody [ESEE122]) has been tested to work in IHC-Fr. Although the other antibodies may work in with this condition they have not been tested as such and therefore I cannot be confident that this product will meet your needs completely. As such I would recommend using ab8366 for your experiments. Positive controls for this product include glioblastoma multiformae and hypoxia-induced human placenta. I would also like to highly recommend ab1 (Anti-HIF1 alpha antibody [H1alpha67] - ChIP Grade). This product has been tested in IHC-IF on mouse and has been referenced in 33 publications. The Abreviews for this product may also be an excellent resource for your experiments. I've included links to both these antibodies as well as the direct link to the IHC-Fr Abreviews for Ab1 below. ab8366: https://www.abcam.com/hif1-alpha-antibody-esee122-ab8366.html ab1: https://www.abcam.com/HIF1-alpha-antibody-H1alpha67-ChIP-Grade-ab1.html ab1 reviews: https://www.abcam.com/HIF-1-alpha-antibody-H1alpha67-ab1.html&tab=abreviews&intabreviewid=9511 I hope this information helps you. I would also like you to know that these products are covered by our Abpromise®. Which ensures that you can trust our products to perform as stated on the datasheets, or we will offer a replacement, credit, or refund. The Abcam Abpromise® guarantee: • 100% Scientific and Customer Support for any product you buy from Abcam or one of our authorized distributors. • We guarantee our products work in the tested species and applications stated on the datasheet. • We will replace or refund products not performing as stated on the datasheet if reported within 6 months of purchase. • We investigate any quality concerns raised by customers to ensure our catalog contains products that perform to the highest standards. Please note these conditions to our Abpromise®: • Protocol information must be provided in order for the claim to be validated. • Antibodies tested in recombinant samples only are not guaranteed for use on endogenous samples. • “Predicted to react” information on the datasheets is provided for reference only and these species are not guaranteed. • Fast Track antibodies are covered for ELISA against the immunizing peptide only. I hope this information is helpful, but please do not hesitate to contact us with further questions.

      Read More

      Abcam Scientific Support

      Answered on Dec 22 2011

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-HIF1 alpha antibody [ESEE122]

      Good
      Abreviews
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
      Sample
      Human Tissue sections (lung)
      Specification
      lung
      Fixative
      Formaldehyde
      Antigen retrieval step
      Heat mediated - Buffer/Enzyme Used: pH 6 citrate buffer
      Permeabilization
      No
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
      Read More

      Abcam user community

      Verified customer

      Submitted Jun 04 2008

      Question

      Hola buenos dias: Resulta que estoy trabajando con un anticuerpo suyo, concretamente el HIF-1 alpha (ref: ab8366, de NOVUS), dicho anticuerpo lo estamos probando en rata, pero no obtenemos resultados óptimos. Me gustaria si me pueden recomendar alguno de sus anticuerpos de HIF-1 alpha que funcione bien en rata, sobre todo para WB. Tambien, si es posible, me manden una alícuota para probarlo. Muchas gracias

      Read More

      Abcam community

      Verified customer

      Asked on Dec 05 2006

      Answer

      Thank you for your enquiry. I apologize that I am unable to answer you in Spanish. Ab8366 has not yet been tested in Western blot, so it may be this antibody is not suitable for that application. You may wish to submit an Abreview of your results from the antibody datasheet. Plese take a look at ab1 and ab2185, both of which were tested in WB and cross-react with rat. I hope this information helps. Please do not hesitate to contact us if you need anything further.

      Read More

      Abcam Scientific Support

      Answered on Dec 05 2006

      Question

      I have used antibody ab8366 for western blot and I have a strong band at 60 kDa. I would like to ask if the antibody also recognizes a douple band corresponding to a dimer or is it just 120 kDa the expected band.

      Read More

      Abcam community

      Verified customer

      Asked on Jun 19 2003

      Answer

      120 kDa is the expected band but the 60 kDa band is often seen in both hypoxic and normoxic cells. The same band is often seen when using ab1 as well. We do not know what the 60 kDa band represents at this time. However, using nuclear extracts decreases this background band and increases the HIF-1 alpha signal. ***Abcam does not recommend ab8366 in Western blotting ***

      Read More

      Abcam Scientific Support

      Answered on Jun 20 2003

      Question

      In August, we received an anti-body against HIF 1alpha (ab8366). We tested this ab in several differnt western blottings using rat and human endothelial cells (control and 24 hrs hypoxia). None of the blots gave a clear result about the presence of HIF 1alpha. Potocol: boil total protein load and run semi-dry blotting on hybond P or C block with milk powder 5% 1e ab (ab8366) 3 hours 2e anti mouse IgG 45 min. substrate ECL++ kit This results in a laddering pattern of several non-specific bands, without the expected 120 kD band. There is no difference between hypoxia and control samples. Maybe you can give us a few tips for optimising this protocol. Do you know a positive control, wich we can use for this western blot? Futher more, I received an e-mail about a new anti-body against HIF1 alpha, especially for western blotting. Could it be possible that we can test this ab.

      Read More

      Abcam community

      Verified customer

      Asked on Dec 17 2002

      Answer

      Generally speaking, we recommend NB 100-131 (ab8366) for immunohistochemistry work. It has been used for western analyis on human and rat samples (MCF7 and PC12 nuclear extracts). The other antibody of which you speak is ab2185, a polyclonal made to the same fusion protein as ab1 and ab463. It has been tested on the same samples as those mentioned above. ***Update 2006: we do not recommend ab8366 for Western blotting***

      Read More

      Abcam Scientific Support

      Answered on Dec 31 2002

      Question

      I have just received your HIF-1 alpha antibody (ref. ab8366)and the protocol suggests to use it at 1:8000. I am just enquiring if this incubation is overnight and whether or not it requires any pre-treatments.

      Read More

      Abcam community

      Verified customer

      Asked on Feb 12 2002

      Answer

      The 1:8,000 dilution was done for 90 minutes at RT by some researchers at the University of Oxford with antigen unmasking. It has also been done at 1:1,000 for 40 hrs at 5C (see review for this product) with pre-treatments (Dako). The dilutions and incubation time seem to vary according to the protocol.

      Read More

      Abcam Scientific Support

      Answered on Feb 18 2002

      Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
      For licensing inquiries, please contact partnerships@abcam.com

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