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  1. Link

    aif-antibody-mitochondrial-marker-ab1998.pdf

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Cell Biology Apoptosis Nucleus Chromatin Condensation
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Anti-AIF 抗体 - Mitochondrial Marker (ab1998)

  • Datasheet
Reviews (3)Q&A (5)References (36)

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Western blot - Anti-AIF antibody - Mitochondrial Marker (ab1998)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AIF antibody - Mitochondrial Marker (ab1998)
  • Western blot - Anti-AIF antibody - Mitochondrial Marker (ab1998)
  • Western blot - Anti-AIF antibody - Mitochondrial Marker (ab1998)
  • Immunocytochemistry - Anti-AIF antibody - Mitochondrial Marker (ab1998)
  • Immunocytochemistry - Anti-AIF antibody - Mitochondrial Marker (ab1998)

Key features and details

  • Rabbit polyclonal to AIF - Mitochondrial Marker
  • Suitable for: WB, IHC-P, ICC
  • Reacts with: Mouse, Rat, Human
  • Isotype: IgG

こちらの製品もご検討ください

タンパク質
Product image
Recombinant Human AIF protein (ab177656)
二次抗体
Product image
Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
ノックアウト
Product image
Human AIFM1 (AIF) knockout HEK-293T cell line (ab266347)

関連製品

製品の概要

  • 製品名

    Anti-AIF antibody - Mitochondrial Marker
    AIF 一次抗体 製品一覧
  • 製品の詳細

    Rabbit polyclonal to AIF - Mitochondrial Marker
  • 由来種

    Rabbit
  • アプリケーション

    適用あり: WB, IHC-P, ICCmore details
  • 種交差性

    交差種: Mouse, Rat, Human
  • 免疫原

    Synthetic peptide corresponding to Human AIF aa 500-600.
    Database link: O95831-1

  • ポジティブ・コントロール

    • WB: Human Caco-2, Daudi, HeLa, HepG2, MCF7, NIH3T3, YB2/0 and K562 cell lysate. Rat and mouse heart lysate. IHC-P: Human retina. ICC: Human U2OS and HeLa cells.
  • 特記事項

    Apoptosis Inducing Factor

    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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

製品の特性

  • 製品の状態

    Liquid
  • 保存方法

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Store at -20°C. Avoid freeze / thaw cycle. Stable for 12 months at -20°C.
  • バッファー

    pH: 7.2
    Preservative: 0.02% Sodium azide
  • Concentration information loading...
  • 精製度

    Affinity purified
  • 特記事項(精製)

    AIF Antibody is affinity chromatography purified via peptide column.
  • ポリ/モノ

    ポリクローナル
  • アイソタイプ

    IgG
  • 研究分野

    • Cell Biology
    • Apoptosis
    • Nucleus
    • Chromatin Condensation
    • Epigenetics and Nuclear Signaling
    • Cell cycle
    • Apoptosis
    • Nuclear
    • Cancer
    • Invasion/microenvironment
    • Apoptosis
    • Chromatin condensation
    • Metabolism
    • Pathways and Processes
    • Redox metabolism
    • Oxidative stress
    • Metabolism
    • Pathways and Processes
    • Metabolism processes
    • Apoptosis
    • Cancer
    • Cell Death
    • Apoptosis
    • Nucleus
    • Chromatin condensation
    • Cancer
    • Cell Death
    • Apoptosis
    • Metabolism
    • Neuroscience
    • Processes

関連製品

  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Positive Controls

    • K-562 whole cell lysate (ab7911)
  • Recombinant Protein

    • Recombinant Human AIF protein (ab177656)

アプリケーション

The Abpromise guarantee

Abpromise保証は、 次のテスト済みアプリケーションにおけるab1998の使用に適用されます

アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。

アプリケーション Abreviews 特記事項
WB (2)
Use a concentration of 0.25 - 1 µg/ml. Detects a band of approximately 67 kDa (predicted molecular weight: 67 kDa). Can be blocked with AIF (internal) peptide (human).
IHC-P (1)
Use at an assay dependent concentration.
ICC
Use a concentration of 1 µg/ml.
特記事項
WB
Use a concentration of 0.25 - 1 µg/ml. Detects a band of approximately 67 kDa (predicted molecular weight: 67 kDa). Can be blocked with AIF (internal) peptide (human).
IHC-P
Use at an assay dependent concentration.
ICC
Use a concentration of 1 µg/ml.

ターゲット情報

  • 機能

    Probable oxidoreductase that has a dual role in controlling cellular life and death; during apoptosis, it is translocated from the mitochondria to the nucleus to function as a proapoptotic factor in a caspase-independent pathway, while in normal mitochondria, it functions as an antiapoptotic factor via its oxidoreductase activity. The soluble form (AIFsol) found in the nucleus induces 'parthanatos' i.e., caspase-independent fragmentation of chromosomal DNA. Interacts with EIF3G,and thereby inhibits the EIF3 machinery and protein synthesis, and activates casapse-7 to amplify apoptosis. Plays a critical role in caspase-independent, pyknotic cell death in hydrogen peroxide-exposed cells. Binds to DNA in a sequence-independent manner.
  • 関連疾患

    Defects in AIFM1 are the cause of combined oxidative phosphorylation deficiency type 6 (COXPD6) [MIM:300816]. It is a mitochondrial disease resulting in a neurodegenerative disorder characterized by psychomotor delay, hypotonia, areflexia, muscle weakness and wasting.
  • 配列類似性

    Belongs to the FAD-dependent oxidoreductase family.
  • 翻訳後修飾

    Under normal conditions, a 54-residue N-terminal segment is first proteolytically removed during or just after translocation into the mitochondrial intermembrane space (IMS) by the mitochondrial processing peptidase (MPP) to form the inner-membrane-anchored mature form (AIFmit). During apoptosis, it is further proteolytically processed at amino-acid position 101 leading to the generation of the mature form, which is confined to the mitochondrial IMS in a soluble form (AIFsol). AIFsol is released to the cytoplasm in response to specific death signals, and translocated to the nucleus, where it induces nuclear apoptosis in a caspase-independent manner.
  • 細胞内局在

    Mitochondrion intermembrane space. Mitochondrion inner membrane. Cytoplasm. Nucleus. Cytoplasm > perinuclear region. Proteolytic cleavage during or just after translocation into the mitochondrial intermembrane space (IMS) results in the formation of an inner-membrane-anchored mature form (AIFmit). During apoptosis, further proteolytic processing leads to a mature form, which is confined to the mitochondrial IMS in a soluble form (AIFsol). AIFsol is released to the cytoplasm in response to specific death signals, and translocated to the nucleus, where it induces nuclear apoptosis. Colocalizes with EIF3G in the nucleus and perinuclear region.
  • Target information above from: UniProt accession O95831 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 参照データベース

    • Entrez Gene: 51060 Human
    • Entrez Gene: 9131 Human
    • Entrez Gene: 26926 Mouse
    • Entrez Gene: 83533 Rat
    • Omim: 300169 Human
    • SwissProt: O95831 Human
    • SwissProt: Q9Z0X1 Mouse
    • SwissProt: Q9JM53 Rat
    • Unigene: 424932 Human
    • Unigene: 476033 Human
    • Unigene: 240434 Mouse
    • Unigene: 203165 Rat
    see all
  • 別名

    • AIFM1 antibody
    • AIFM1_HUMAN antibody
    • Apoptosis inducing factor 1, mitochondrial antibody
    • Apoptosis inducing factor antibody
    • Apoptosis inducing factor, mitochondrion associated, 1 antibody
    • Apoptosis-inducing factor 1 antibody
    • CMTX4 antibody
    • COWCK antibody
    • COXPD6 antibody
    • Harlequin antibody
    • Hq antibody
    • mAIF antibody
    • MGC111425 antibody
    • MGC5706 antibody
    • mitochondrial antibody
    • Neuropathy, axonal motor-sensory, with deafness and mental retardation antibody
    • neuropathy, axonal, motor-sensory with deafness and mental retardation (Cowchock syndrome) antibody
    • PDCD 8 antibody
    • PDCD8 antibody
    • Programmed cell death 8 (apoptosis inducing factor) antibody
    • Programmed cell death 8 antibody
    • Programmed cell death 8 isoform 1 antibody
    • Programmed cell death 8 isoform 2 antibody
    • Programmed cell death 8 isoform 3 antibody
    • Programmed cell death protein 8 antibody
    • Programmed cell death protein 8 mitochondrial antibody
    • Programmed cell death protein 8 mitochondrial precursor antibody
    • Programmed cell death protein 8 mitochondrial precursor antibody
    • Striatal apoptosis inducing factor antibody
    see all

画像

  • Western blot - Anti-AIF antibody - Mitochondrial Marker (ab1998)
    Western blot - Anti-AIF antibody - Mitochondrial Marker (ab1998)
    All lanes : Anti-AIF antibody - Mitochondrial Marker (ab1998) at 1 µg/ml

    Lane 1 : K562 cell lysate
    Lane 2 : Rat heart tissue lysate
    Lane 3 : Mouse heart tissue lysate

    Predicted band size: 67 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AIF antibody - Mitochondrial Marker (ab1998)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-AIF antibody - Mitochondrial Marker (ab1998)
    Immunohistochemistry of AIF in human retina with anti-AIF (IN) at 10 µg/ml.
  • Western blot - Anti-AIF antibody - Mitochondrial Marker (ab1998)
    Western blot - Anti-AIF antibody - Mitochondrial Marker (ab1998)
    All lanes : Anti-AIF antibody - Mitochondrial Marker (ab1998) at 1 µg/ml

    Lane 1 : Caco-2 cell lysate
    Lane 2 : Daudi cell lysate
    Lane 3 : HeLa cell lysate
    Lane 4 : HepG2 cell lysate
    Lane 5 : K562 cell lysate
    Lane 6 : MCF7 cell lysate

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit IgG HRP conjugate at 1/10000 dilution

    Predicted band size: 67 kDa

  • Western blot - Anti-AIF antibody - Mitochondrial Marker (ab1998)
    Western blot - Anti-AIF antibody - Mitochondrial Marker (ab1998)
    All lanes : Anti-AIF antibody - Mitochondrial Marker (ab1998) at 1 µg/ml

    Lane 1 : NIH3T3 cell lysate
    Lane 2 : YB2/0 cell lysate

    Lysates/proteins at 15 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit IgG HRP conjugate at 1/10000 dilution

    Predicted band size: 67 kDa

  • Immunocytochemistry - Anti-AIF antibody - Mitochondrial Marker (ab1998)
    Immunocytochemistry - Anti-AIF antibody - Mitochondrial Marker (ab1998)Image from Varecha M et al, J Biomed Sci. 2009 Jul 6;16:59, fig 2.

    ab1998 staining AIF in human U2OS cells by Immunocytochemistry. Samples were fixed using 4% paraformaldehyde.Left image shows fixed cells labeled with ab1998 (red) and cyclophilin A (green).
    Right image shows U2OS cell 6 hours after induction of apoptosis by 200 nM staurosporine.Note translocated of AIF to the nucleus upon induction of apoptosis.

  • Immunocytochemistry - Anti-AIF antibody - Mitochondrial Marker (ab1998)
    Immunocytochemistry - Anti-AIF antibody - Mitochondrial Marker (ab1998)

    ICC image of ab1998 stained HeLa cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab1998, 1µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

プロトコール

  • Western blot protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

データシートおよび資料

  • Datasheet download

    Download

参考文献 (36)

ab1998 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab1998 は 36 報の論文で使用されています。

  • Liu C  et al. Metformin revert insulin-induced oxaliplatin resistance by activating mitochondrial apoptosis pathway in human colon cancer HCT116 cells. Cancer Med 9:3875-3884 (2020). PubMed: 32248666
  • Power MJ  et al. Systematic spatiotemporal mapping reveals divergent cell death pathways in three mouse models of hereditary retinal degeneration. J Comp Neurol 528:1113-1139 (2020). PubMed: 31710697
  • Marchi S  et al. Akt-mediated phosphorylation of MICU1 regulates mitochondrial Ca2+ levels and tumor growth. EMBO J 38:N/A (2019). PubMed: 30504268
  • Qin QJ  et al. Rhynchophylline ameliorates myocardial ischemia/reperfusion injury through the modulation of mitochondrial mechanisms to mediate myocardial apoptosis. Mol Med Rep 19:2581-2590 (2019). PubMed: 30720139
  • Zhang C  et al. Curcumin induces apoptosis and inhibits angiogenesis in murine malignant mesothelioma. Int J Oncol 53:2531-2541 (2018). PubMed: 30272283
View all Publications for this product

レビューと Q&A

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レビューを送る 質問を送る

1-8 of 8 Abreviews or Q&A

Western blot abreview for Anti-AIF antibody - Mitochondrial Marker

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (MEF)
Gel Running Conditions
Reduced Denaturing
Loading amount
20 µg
Treatment
100 U/ml IL-1
Specification
MEF
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 Jun 04 2021

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-AIF antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Breast cancer tissue and adjacent normal)
Specification
Breast cancer tissue and adjacent normal
Fixative
Formaldehyde
Antigen retrieval step
Other
Permeabilization
No
Blocking step
H2o2 of abcam secondary antibody Kit Expose Kit as blocking agent for 10 minute(s) · Concentration: 1% · Temperature: 37°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

MRS. Thushara Jayan Pillai

Verified customer

投稿 Oct 24 2011

Western blot abreview for Anti-AIF antibody - Mitochondrial Marker

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (SW1573 lung cancer cell line)
Gel Running Conditions
Reduced Denaturing (10% SDS-PAGE)
Loading amount
50 µg
Specification
SW1573 lung cancer cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 8% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

投稿 May 27 2011

Question

I am interested in looking at AIF as a potential marker of caspase-independent apoptosis. Please can you tell me if this is a reliable marker or if there is another product you would recommend? I don’t see any caspase cleavage but I do see mitochondrial membrane depolarisation and membrane blebbing in my experiments suggesting that some other form of death is occurring.

Also, to use it for this purpose would it be best to look for it in mitochondrial and nuclear fractions by immunoblotting as I presume that FACS & standard immunoblotting would only tell you if it was present in the cell or not or do levels increase after mitochondrial release? Would your cellular fractionation kit ab109719 be suitable for looking at AIF in these fractions?

I am working with rat cells, which of your antibodies is best for this cell type: ab119917, ab32516 or ab1998?

Sorry this is not an area that I am familiar with!

Thanks

Read More

Abcam community

Verified customer

Asked on Aug 15 2012

Answer

Thank you for contacting us.

I would suggested searching the potential of this as caspase-independent apoptosis marker in publications. There are many publications related to this protein. Please have a look in pubmed.

As the process triggering apoptosis involves the mitochondrial release of AIF through the mitochondrial membrane, which enters the cytosol, and finally ends up in the cell nucleus where it signals the cell to condense its chromosomes and fragment its DNA molecules in order to prepare for cell death. So the idea would be to compare the AIF level in mitochondrial, cytosolic and nuclear preparations. The kit ab109719 should be able to separate the mitochondrial, cytosolic and nuclear fractions for experiment.

Monoclonal antibodies provide clean WB results so monoclonal will be best suitable e.g. ab119917 and ab32516. This however does not mean polyclonal aren't good, they are good but monoclonal are better.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

Read More

Abcam Scientific Support

Answered on Aug 16 2012

Question

i am encountering a problem with finding the peptide sequences of THE BELOW ON ABCAM WEBSITE ;

Rabbit polyclonal to AIF
ab 1998

Rabbit polyclonal to Bcl2
ab 59348

Rabbit monoclonal [E83-77] to active
Caspase 3
ab 32042

CAN YOU PLS SEND ME THE PEPTIDE SEQUENCES OF THE ABOVE PLS I NEED THEM FOR THE FURTHER STUDIES.

Read More

Abcam community

Verified customer

Asked on May 04 2012

Answer



ab1998 : https://www.abcam.com/ab1998, where the immunogen is described asa peptide corresponding to amino acids 517 to 531 of human AIF. This sequence is identical tothose of mouse and rat AIF.

ab32042 : https://www.abcam.com/ab32042, where the immunogen is described as : a synthetic peptide corresponding to residues following Ser29 of human Caspase 3 (N terminus of p17 subunit).

ab59348 : https://www.abcam.com/ab59348, where the immunogen is described as a synthesized non phosphopeptide derived from human Bcl2 around the phosphorylation site of threonine 69 (ARTPSP).

Read More

Abcam Scientific Support

Answered on May 04 2012

Question

I´ve been following this problem, even though in this particular Ab I´m not the final user. First of all the concentration the data sheet suggested is the one used from the beginning, also incubation times has been tested ( 1 hour at RT or ON at 4 ºC) so the problem is not that we have a weak band , it is that there is no band at all (even at 4 ºC ON) so we believe your Ab is not working. On the other hand we bought at the same time the COX IV loading control which works fine at the concentration suggested on the data sheet and 1 hour at RT. Increasing Ab concentrations is would be possible if we see a weak signal...remember we have not signal even at ON incubations and it is different to data sheet suggestions and what we expected on this Ab at the time we choose it. More over Carlos thesis work has been done mainly using western blot experiments on same model so I´m clear that we have not problems on the technique.

Read More

Abcam community

Verified customer

Asked on Dec 21 2005

Answer

Thank you for your email. In your prior message with the attachment containing the Western blot image, incubation for 1 hr at RT was mentioned but not overnight at 4C. I agree with you and I would expect that you would see some sort of signal with your samples and overnight incubation. As we guarantee our products to work as stated on the online datasheet, I would be happy to offer you a free of charge replacement vial or a credit note or refund. Please let me know how you would like to proceed, and happy holidays!

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Abcam Scientific Support

Answered on Dec 22 2005

Question

BATCH NUMBER 108063 ORDER NUMBER 112828 DESCRIPTION OF THE PROBLEM No signal (No band at all) SAMPLE Purified Rat liver Mitochondria PRIMARY ANTIBODY AB 1998 DETECTION METHOD Super signal west pico (pierce) POSITIVE AND NEGATIVE CONTROLS USED We have tried just rat liver mitochondria lysates using triton x 100, SDS or NP 40. This is my model, and the release of cytochrome c works fine, so I am not interested in other species. ANTIBODY STORAGE CONDITIONS The antibody was Aliquoted upon arrival and store at -30 ?C SAMPLE PREPARATION lysis buffer containing PMSF 1 mM, EDTA 10 mM, Nacl 300 mM, BME 5 mM, NP40 1 %,Tris 50 mM (pH: 7.6) plus protease inhibitors cocktail. AMOUNT OF PROTEIN LOADED 5-30 ?g per lane ELECTROPHORESIS/GEL CONDITIONS Std 12 %SDS-PAGE, run at 170 volts during 1 hour. TRANSFER AND BLOCKING CONDITIONS Wet transfer, 25 volts over night. Blocking solution: 5 % Non fat milk in TBS. 1 Hour RT. SECONDARY ANTIBODY Goat anti-Rabbit- HRP conjugated (AP 132P, Chemicon) HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 HAVE YOU RUN A "NO PRIMARY" CONTROL? No WHAT STEPS HAVE YOU ALTERED? NONE ADDITIONAL NOTES I have experience with several anti-bodies. For most of them the western blot condition are the same with not troubles of any kind. I am investigating the release of apoptotic factors from isolated mitochondria induced by Bax. The detection in this experiments of cytochrome c works fine. But now I wanted to try AIF release in this model. But the AB 1998 anti-body did not work.

Read More

Abcam community

Verified customer

Asked on Dec 20 2005

Answer

Thank you for your enquiry and I'm sorry to hear that you are experiencing difficulty with ab1998. At this time I would like to make the following suggestions in order to help improve your results. I would suggest extending the incubation period with the primary antibody to overnight at 4C and also increasing the concentration of ab1998 to 1:500. I also suggest running a positive control - K562 cell lysate is recommended for ab1998. Please contact us again if you have any additional questions, and happy holidays.

Read More

Abcam Scientific Support

Answered on Dec 21 2005

Question

Hello! I would like use the rabbit polyclonal AIF antibody (ab1998 or ab2086) on paraffin sections 5-8 micron. Do you have some experience on this possibility?; can you give me some suggestions?

Read More

Abcam community

Verified customer

Asked on Sep 06 2002

Answer

This antibody has not been tested for IHC, therefore we cannot guarantee results.

Read More

Abcam Scientific Support

Answered on Sep 09 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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製品・研究分野別
  • 癌
  • 循環器
  • 細胞生物学
  • 発生
  • エピジェネティクス&核内情報
  • 免疫学
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