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Cardiovascular Hypoxia HIF

Anti-HIF1 beta抗体[2B10] (ab2771)

  • Datasheet
  • SDS
Reviews (7)Q&A (5)References (18)

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Immunocytochemistry/ Immunofluorescence - Anti-HIF1 beta antibody [2B10] (ab2771)
  • Immunocytochemistry/ Immunofluorescence - Anti-HIF1 beta antibody [2B10] (ab2771)
  • Immunocytochemistry/ Immunofluorescence - Anti-HIF1 beta antibody [2B10] (ab2771)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF1 beta antibody [2B10] (ab2771)
  • Flow Cytometry - Anti-HIF1 beta antibody [2B10] (ab2771)

Key features and details

  • Mouse monoclonal [2B10] to HIF1 beta
  • Suitable for: Flow Cyt, WB, ICC/IF, IHC-P, IP
  • Reacts with: Mouse, Rat, Human, Xenopus laevis, Fish, Non human primates, Zebrafish, African green monkey
  • Isotype: IgG1

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概述

  • 产品名称

    Anti-HIF1 beta抗体[2B10]
    参阅全部 HIF1 beta 一抗
  • 描述

    小鼠单克隆抗体[2B10] to HIF1 beta
  • 宿主

    Mouse
  • 特异性

    Detects aryl hydrocarbon (Ah) receptor nuclear translocator (ARNT).
  • 经测试应用

    适用于: Flow Cyt, WB, ICC/IF, IHC-P, IPmore details
  • 种属反应性

    与反应: Mouse, Rat, Human, Xenopus laevis, Fish, Non human primates, Zebrafish, African green monkey
    预测可用于: Rabbit, Cow
  • 免疫原

    Synthetic peptide corresponding to Human HIF1 beta aa 771-789.
    Sequence:

    NSYNNEEFPDLTMFPPFSE

    Run BLAST with BLAST the sequence with ExPASy Run BLAST with BLAST the sequence with NCBI
  • 常规说明

    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

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
  • 存储溶液

    Preservative: 0.05% Sodium azide
    Constituents: 0.1% BSA, 99% PBS
  • Concentration information loading...
  • 纯度

    Affinity purified
  • 克隆

    单克隆
  • 克隆编号

    2B10
  • 同种型

    IgG1
  • 研究领域

    • 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

相关产品

  • 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)
  • Conjugation kits

    • PE / R-Phycoerythrin Conjugation Kit - Lightning-Link® (ab102918)
    • APC Conjugation Kit - Lightning-Link® (ab201807)
  • Isotype control

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

    • Recombinant Human HIF1 beta protein (ab157907)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab2771于以下的经测试应用

“应用说明”部分 下显示的仅为推荐的起始稀释度;实际最佳的稀释度/浓度应由使用者检定。

应用 Ab评论 说明
Flow Cyt
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB (6)
Use at an assay dependent concentration.
EMSA
Use at an assay dependent concentration.
Gel Shift Assay
Use at an assay dependent concentration.
ICC/IF
1/1000.
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP (1)
Use at an assay dependent concentration.
说明
Flow Cyt
Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

WB
Use at an assay dependent concentration.
EMSA
Use at an assay dependent concentration.
Gel Shift Assay
Use at an assay dependent concentration.
ICC/IF
1/1000.
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
Use at an assay dependent concentration.

靶标

  • 功能

    Required for activity of the Ah (dioxin) receptor. This protein is required for the ligand-binding subunit to translocate from the cytosol to the nucleus after ligand binding. The complex then initiates transcription of genes involved in the activation of PAH procarcinogens. The heterodimer with HIF1A or EPAS1/HIF2A functions as a transcriptional regulator of the adaptive response to hypoxia.
  • 序列相似性

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

    Nucleus.
  • Target information above from: UniProt accession P27540 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • 数据库链接

    • Entrez Gene: 281010 Cow
    • Entrez Gene: 405 Human
    • Entrez Gene: 11863 Mouse
    • Entrez Gene: 100008996 Rabbit
    • Entrez Gene: 25242 Rat
    • Entrez Gene: 64277 Zebrafish
    • Omim: 126110 Human
    • SwissProt: Q9BE97 Cow
    • SwissProt: P27540 Human
    • SwissProt: P53762 Mouse
    • SwissProt: O02748 Rabbit
    • SwissProt: P41739 Rat
    • Unigene: 632446 Human
    • Unigene: 250265 Mouse
    • Unigene: 10520 Rat
    see all
  • 别名

    • arnT antibody
    • ARNT protein antibody
    • ARNT_HUMAN antibody
    • Aryl hydrocarbon receptor nuclear translocator antibody
    • bHLHe2 antibody
    • Class E basic helix-loop-helix protein 2 antibody
    • Dioxin receptor antibody
    • Dioxin receptor nuclear translocator antibody
    • Drnt antibody
    • HIF 1 beta antibody
    • HIF 1beta antibody
    • HIF-1-beta antibody
    • HIF1-beta antibody
    • HIF1B antibody
    • HIF1beta antibody
    • Hypoxia Inducible Factor 1 antibody
    • Hypoxia inducible factor 1 beta antibody
    • Hypoxia-inducible factor 1-beta antibody
    • nuclear translocator antibody
    • Tango antibody
    see all

图片

  • Immunocytochemistry/ Immunofluorescence - Anti-HIF1 beta antibody [2B10] (ab2771)
    Immunocytochemistry/ Immunofluorescence - Anti-HIF1 beta antibody [2B10] (ab2771)

    Immunocytochemistry/Immunofluorescence analysis of HIF1 beta shows staining in A2058 cells. HIF1 beta (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2771 (1:100) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/ Immunofluorescence - Anti-HIF1 beta antibody [2B10] (ab2771)
    Immunocytochemistry/ Immunofluorescence - Anti-HIF1 beta antibody [2B10] (ab2771)

    Immunocytochemistry/Immunofluorescence analysis of HIF1 beta shows staining in HeLa cells. HIF1 beta (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2771 (1:20) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunocytochemistry/ Immunofluorescence - Anti-HIF1 beta antibody [2B10] (ab2771)
    Immunocytochemistry/ Immunofluorescence - Anti-HIF1 beta antibody [2B10] (ab2771)

    Immunocytochemistry/Immunofluorescence analysis of HIF1 beta shows staining in U251 cells. HIF1 beta (green), F-Actin staining with Phalloidin (red) and nuclei with DAPI (blue) is shown. Cells were grown on chamber slides and fixed with formaldehyde prior to staining. Cells were probed without (control) or with ab2771 (1:100) overnight at 4°C, washed with PBS and incubated with a DyLight-488 conjugated secondary antibody. Images were taken at 60X magnification.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF1 beta antibody [2B10] (ab2771)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-HIF1 beta antibody [2B10] (ab2771)
    IHC image of ab2771 staining in human tonsil formalin fixed paraffin embedded tissue section, performed on a Leica BondTM 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 ab2771, 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.

  • Flow Cytometry - Anti-HIF1 beta antibody [2B10] (ab2771)
    Flow Cytometry - Anti-HIF1 beta antibody [2B10] (ab2771)
    Overlay histogram showing PC12 cells stained with ab2771 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab2771, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in PC12 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

实验方案

  • Flow cytometry protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (18)

发表研究结果有使用 ab2771?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab2771 被引用在 18 文献中.

  • Wu CS  et al. ASC modulates HIF-1a stability and induces cell mobility in OSCC. Cell Death Dis 11:721 (2020). PubMed: 32883954
  • Zhao Y  et al. Aryl hydrocarbon receptor nuclear translocator promotes the proliferation and invasion of clear cell renal cell carcinoma cells potentially by affecting the glycolytic pathway. Oncol Lett 20:56 (2020). PubMed: 32793310
  • Li C  et al. microRNA-222-Mediated VHL Downregulation Facilitates Retinoblastoma Chemoresistance by Increasing HIF1a Expression. Invest Ophthalmol Vis Sci 61:9 (2020). PubMed: 32756923
  • Fu YC & Xin ZM Inhibited corneal neovascularization in rabbits following corneal alkali burn by double-target interference for VEGF and HIF-1a. Biosci Rep 39:N/A (2019). PubMed: 30355648
  • Chen WC  et al. Aryl hydrocarbon receptor modulates stroke-induced astrogliosis and neurogenesis in the adult mouse brain. J Neuroinflammation 16:187 (2019). PubMed: 31606043
View all Publications for this product

客户评价及客户问答

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1-10 of 12 Abreviews or Q&A

Western blot abreview for Anti-HIF1 beta antibody [2B10]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HOS cells)
Loading amount
50 µg
Specification
HOS cells
Gel Running Conditions
Reduced Denaturing (10%)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°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

提交于 Mar 03 2009

Immunoprecipitation abreview for Anti-HIF1 beta antibody [2B10]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunoprecipitation
Sample
Human Cell lysate - whole cell (293T)
Total protein in input
500 µg
Specification
293T
Immuno-precipitation step
Protein G
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

提交于 Oct 30 2007

Western blot abreview for Anti-HIF1 beta antibody [2B10]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
African Green Monkey Cell lysate - whole cell (Cos-7 cell line)
Loading amount
30 µg
Specification
Cos-7 cell line
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°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

提交于 Sep 26 2007

Western blot abreview for Anti-HIF1 beta antibody [2B10]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (3T3 cell line)
Loading amount
30 µg
Specification
3T3 cell line
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 21°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

提交于 Sep 26 2007

Western blot abreview for Anti-HIF1 beta antibody [2B10]

Average
Abreviews
Abreviews
Application
Western blot
Sample
Mouse Tissue lysate - whole (forebrain)
Loading amount
15 µg
Specification
forebrain
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 15 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°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

提交于 Jun 22 2007

Western blot abreview for Anti-HIF1 beta antibody [2B10]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Rat Cell lysate - whole cell (PC12)
Loading amount
20 µg
Specification
PC12
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 21°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

提交于 Jun 21 2007

Western blot abreview for Anti-HIF1 beta antibody [2B10]

Good
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (U2OS)
Loading amount
20 µg
Specification
U2OS
Gel Running Conditions
Reduced Denaturing
Blocking step
Milk as blocking agent for 45 minute(s) · Concentration: 5% · Temperature: 21°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

提交于 Jun 15 2007

Question

Inquiry: I am just wondering if this antibody can react with mouse brain homogenates? Do you have any literature to show the result? Thanks a lot!

Read More

Abcam community

Verified customer

Asked on Jun 10 2013

Answer

While we do have publications using this antibody in mouse samples we are not presently aware of specific publications using mouse brain homogenates. HIF1 beta forms heterodimers with HIF1alpha and has ubiquitous nuclear expression. The HIF1 complex rapidly degrades. It is critical to prep only a few plates/dishes/flasks of cells at a time and to immediately get the cells into ice cold buffers and perform the whole protein prep on ice. HIF-1 is largely undetectable in cells or tissues grown under normoxic conditions. It is stabilized only at O2 concentrations below 5% or with treat using certain agents (CoCl2, DFO, etc.) so proper sample preparation is critical. Upon stabilization HIF-1 translocates to the nucleus. The best western blots (cleanest) are always done using nuclear extracts. It is possible to detect HIF-1 in whole cell extracts, but they tend to be much dirtier and the staining is much weaker. We recommend that a positive/negative control always be run side by side so that it is possible to discern which band is upregulated in the hypoxic sample. Depending on the sample, treatment, etc. you may see either a band or a doublet.

Read More

Abcam Scientific Support

回复于 Jun 10 2013

Question

BATCH NUMBER 271685 ORDER NUMBER -- NOT SPECIFIED -- DESCRIPTION OF THE PROBLEM Double bands, MW is around 90.For human and rat samples, the bands from homogenate are Darker than from extract opposite for mouse samples. The bands from mouse samples are much darker than the others. SAMPLE I tested the antibody with brain samples such as human homogenate, human extract, rat homogenate, rat extract, mouse homogenate and mouse extract. PRIMARY ANTIBODY HIF1 beta ab2771, 271685. 1:1000 diluted, in 5% non-fat milk incubate for 18 hours,overnight, room temperature wash with 0.1% tween-TBS for 3 times, at 10 minutes each time DETECTION METHOD ECL POSITIVE AND NEGATIVE CONTROLS USED not used ANTIBODY STORAGE CONDITIONS -20, but +4 degrees before use diluted (1:4) in 5%BSA. SAMPLE PREPARATION in SDS buffer containing 2%SDS, 10%glycerol, 1%beta-mercaptoethanol and 0.0625M Tris-Cl AMOUNT OF PROTEIN LOADED 10ug ELECTROPHORESIS/GEL CONDITIONS 10% reducing TRANSFER AND BLOCKING CONDITIONS buffer:1.4% glycine, 0.3% Trizma base, 20% methanol 1 hour block with 5% non-fat milk SECONDARY ANTIBODY goat anti-mouse 1:5000 diluted in 5% non-fat milk for 2 hours at room temperature wash with 0.1% tween-TBS for 3 times, at 10 minutes each time HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 2 HAVE YOU RUN A "NO PRIMARY" CONTROL? No DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? not altered ADDITIONAL NOTES I checked literature and found HIF1 beta has several isoforms. Does ab2771 recognize only one isoforms or all? I failed to attach the image of the film, which is jpg or doc format.

Read More

Abcam community

Verified customer

Asked on Jul 30 2007

Answer

Thank you for your patience. I am sorry to hear that you are experiencing difficulties with this product ab2771 in western blot. Often it is possible to make suggestions that help resolve problems. We will happily offer technical support and in the event that a product is not functioning in the applications cited on the product data sheet (and the problem has been reported within 120 days of purchase), and if it appears that the antibody is at fault, a replacement/credit note/refund will be offered. I have looked through the protocols you used and have a few questions and suggestions that might help you resolve the problem. In most cases, the cause of multiple bands is because the primary and/or secondary antibody concentration is too high and causes non-specific binding. You can try decreasing the primary (1:1000 ~ 1:5000) and secondary concentration or run a no-primary control (without the primary antibody) to ensure that the multiple bands are not caused by your secondary antibody. Another possible reason is that the extra bands you are observing may be multimers. Please try boiling your protein in SDS-PAGE for 10 minutes to disrupt multimers. Meanwhile, bands located below the expected molecular weight you are observing may indicate that your target protein has been digested. Please make sure that you incorporate sufficient protease inhibitors in your sample buffer. The image you provided showed different strengths in expression. I would suggest doing optimization for each species sample separately, so that you can have better control of the level and intensity of the bands. This is because your target protein may exist in different amount in different species. I would also suggest incubating your primary antibody at 4 degrees instead of at room temperature. Incubation at room temperature is recommended to be done for 1-2 hours, while 4 degrees can be done overnight. Unfortunately, I do not have any data suggesting that this product can detect other HIF-1 beta isoforms at this time. However, I believe the following reference may help explain the double band that you are observing. Rescue of Hypoxia-inducible Factor-1 alpha-deficient Tumor Growth by Wild-Type Cells Is Independent of Vascular Endothelial Growth Factor. Can. Res. 62(10):2962-2970, 2002. http://cancerres.aacrjournals.org/cgi/reprint/62/10/2962 In this specific reference, the authors detect nonspecific cross-reactivity of the anti-ARNT polyclonal antibody in nuclear extracts of ES cells. Also, we see similar reactivity in the reference: Isoform specific expression of hypoxia-inducible factor-1alpha during the late stages of mouse spermiogenesis. Mol. Endocrinol., Vol 16: 234, Feb 2002. http://mend.endojournals.org/cgi/reprint/16/2/234 I hope the above suggestions and references will be helpful to you. If you still experience problems please do not hesitate to contact me with details of your order (purchase order number, shipping address/purchasing agent info, contact information, etc.) so that I can immediately arrange for a replacement or refund to you. Also, please advice on how you would to proceed with this enquiry. I look forward to hearing from you again in order to resolve the matter.

Read More

Abcam Scientific Support

回复于 Jul 30 2007

Question

I have an inquiry about your product, ab2771; HIF1 beta antibody. Which cell line did you use this antibody for Immunoprecipitation? Was it transfected cells? The researcher wants to use this antibody for IP. And he found clone 2B10 except abcam wouldn't be recommended for IP. Please advise us if you have any infromation about this purpose.

Read More

Abcam community

Verified customer

Asked on Jan 06 2006

Answer

Thank you for your enquiry. Followingf corresponce with the source of this antibody I have determined that the cell lines used for IP were mouse Hep1c1c7 and human HeLa cells. ab2771 works well by IP in both mouse and human lysates at 1-2 ug/IP. I hope this information helps, please do not hesitate to contact us if you need any more advice or information.

Read More

Abcam Scientific Support

回复于 Jan 06 2006

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