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Epigenetics and Nuclear Signaling Histones H3 Methylated

Anti-Histone H3 (tri methyl K79)抗体- ChIP Grade (ab2621)

  • Datasheet
  • SDS
Reviews (11)Q&A (4)References (132)

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ChIP - Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)
  • Western blot - Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)

Key features and details

  • Rabbit polyclonal to Histone H3 (tri methyl K79) - ChIP Grade
  • Suitable for: ChIP, WB
  • Reacts with: Cow, Human
  • Isotype: IgG

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概述

  • 产品名称

    Anti-Histone H3 (tri methyl K79)抗体- ChIP Grade
    参阅全部 Histone H3 一抗
  • 描述

    兔多克隆抗体to Histone H3 (tri methyl K79) - ChIP Grade
  • 宿主

    Rabbit
  • 特异性

    Reacts with Tri-methyl K79 of histone H3. Cross-reactivity to di-methyl K79.
  • 经测试应用

    适用于: ChIP, WBmore details
  • 种属反应性

    与反应: Cow, Human
    预测可用于: Rat, Saccharomyces cerevisiae, Plasmodium falciparum
  • 免疫原

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituent: PBS

    Batches of this product that have a concentration < 1mg/ml may have BSA added as a stabilising agent. If you would like information about the formulation of a specific lot, please contact our scientific support team who will be happy to help.
  • Concentration information loading...
  • 纯度

    Immunogen affinity purified
  • 克隆

    多克隆
  • 同种型

    IgG
  • 研究领域

    • Epigenetics and Nuclear Signaling
    • Histones
    • H3
    • Methylated
    • Epigenetics and Nuclear Signaling
    • ChIP assays
    • ChIP antibodies

相关产品

  • ChIP Related Products

    • Rabbit Anti-Mouse IgG H&L (ab46540)
    • ChIP Kit (ab500)
  • Compatible Secondaries

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

    • Human Histone H3 (di methyl K9) peptide (ab1772)
  • Isotype control

    • Rabbit IgG, polyclonal - Isotype Control (ChIP Grade) (ab171870)
  • Recombinant Protein

    • Recombinant human Histone H3 protein (Active) (ab198757)
  • Related Products

    • Histone H3 (tri-methyl K79) Quantification Kit (Colorimetric) (ab115087)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab2621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
ChIP (4)
Use 2-4 µg for 6 µg of chromatin.
WB (4)
Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).

Abcam recommends using BSA as the blocking agent.

说明
ChIP
Use 2-4 µg for 6 µg of chromatin.
WB
Use a concentration of 1 µg/ml. Detects a band of approximately 17 kDa (predicted molecular weight: 15 kDa).

Abcam recommends using BSA as the blocking agent.

靶标

  • 功能

    Core component of nucleosome. Nucleosomes wrap and compact DNA into chromatin, limiting DNA accessibility to the cellular machineries which require DNA as a template. Histones thereby play a central role in transcription regulation, DNA repair, DNA replication and chromosomal stability. DNA accessibility is regulated via a complex set of post-translational modifications of histones, also called histone code, and nucleosome remodeling.
  • 序列相似性

    Belongs to the histone H3 family.
  • 发展阶段

    Expressed during S phase, then expression strongly decreases as cell division slows down during the process of differentiation.
  • 翻译后修饰

    Acetylation is generally linked to gene activation. Acetylation on Lys-10 (H3K9ac) impairs methylation at Arg-9 (H3R8me2s). Acetylation on Lys-19 (H3K18ac) and Lys-24 (H3K24ac) favors methylation at Arg-18 (H3R17me).
    Citrullination at Arg-9 (H3R8ci) and/or Arg-18 (H3R17ci) by PADI4 impairs methylation and represses transcription.
    Asymmetric dimethylation at Arg-18 (H3R17me2a) by CARM1 is linked to gene activation. Symmetric dimethylation at Arg-9 (H3R8me2s) by PRMT5 is linked to gene repression. Asymmetric dimethylation at Arg-3 (H3R2me2a) by PRMT6 is linked to gene repression and is mutually exclusive with H3 Lys-5 methylation (H3K4me2 and H3K4me3). H3R2me2a is present at the 3' of genes regardless of their transcription state and is enriched on inactive promoters, while it is absent on active promoters.
    Methylation at Lys-5 (H3K4me), Lys-37 (H3K36me) and Lys-80 (H3K79me) are linked to gene activation. Methylation at Lys-5 (H3K4me) facilitates subsequent acetylation of H3 and H4. Methylation at Lys-80 (H3K79me) is associated with DNA double-strand break (DSB) responses and is a specific target for TP53BP1. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are linked to gene repression. Methylation at Lys-10 (H3K9me) is a specific target for HP1 proteins (CBX1, CBX3 and CBX5) and prevents subsequent phosphorylation at Ser-11 (H3S10ph) and acetylation of H3 and H4. Methylation at Lys-5 (H3K4me) and Lys-80 (H3K79me) require preliminary monoubiquitination of H2B at 'Lys-120'. Methylation at Lys-10 (H3K9me) and Lys-28 (H3K27me) are enriched in inactive X chromosome chromatin.
    Phosphorylated at Thr-4 (H3T3ph) by GSG2/haspin during prophase and dephosphorylated during anaphase. Phosphorylation at Ser-11 (H3S10ph) by AURKB is crucial for chromosome condensation and cell-cycle progression during mitosis and meiosis. In addition phosphorylation at Ser-11 (H3S10ph) by RPS6KA4 and RPS6KA5 is important during interphase because it enables the transcription of genes following external stimulation, like mitogens, stress, growth factors or UV irradiation and result in the activation of genes, such as c-fos and c-jun. Phosphorylation at Ser-11 (H3S10ph), which is linked to gene activation, prevents methylation at Lys-10 (H3K9me) but facilitates acetylation of H3 and H4. Phosphorylation at Ser-11 (H3S10ph) by AURKB mediates the dissociation of HP1 proteins (CBX1, CBX3 and CBX5) from heterochromatin. Phosphorylation at Ser-11 (H3S10ph) is also an essential regulatory mechanism for neoplastic cell transformation. Phosphorylated at Ser-29 (H3S28ph) by MLTK isoform 1, RPS6KA5 or AURKB during mitosis or upon ultraviolet B irradiation. Phosphorylation at Thr-7 (H3T6ph) by PRKCBB is a specific tag for epigenetic transcriptional activation that prevents demethylation of Lys-5 (H3K4me) by LSD1/KDM1A. At centromeres, specifically phosphorylated at Thr-12 (H3T11ph) from prophase to early anaphase, by DAPK3 and PKN1. Phosphorylation at Thr-12 (H3T11ph) by PKN1 is a specific tag for epigenetic transcriptional activation that promotes demethylation of Lys-10 (H3K9me) by KDM4C/JMJD2C. Phosphorylation at Tyr-42 (H3Y41ph) by JAK2 promotes exclusion of CBX5 (HP1 alpha) from chromatin.
    Monoubiquitinated by RAG1 in lymphoid cells, monoubiquitination is required for V(D)J recombination (By similarity). Ubiquitinated by the CUL4-DDB-RBX1 complex in response to ultraviolet irradiation. This may weaken the interaction between histones and DNA and facilitate DNA accessibility to repair proteins.
  • 细胞定位

    Nucleus. Chromosome.
  • Target information above from: UniProt accession P68431 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: 8350 Human
    • Entrez Gene: 8351 Human
    • Entrez Gene: 8352 Human
    • Entrez Gene: 8353 Human
    • Entrez Gene: 8354 Human
    • Entrez Gene: 8355 Human
    • Entrez Gene: 8356 Human
    • Entrez Gene: 8357 Human
    • Entrez Gene: 8358 Human
    • Entrez Gene: 8968 Human
    • Entrez Gene: 100364501 Rat
    • Entrez Gene: 100365669 Rat
    • Entrez Gene: 291159 Rat
    • Entrez Gene: 679994 Rat
    • Entrez Gene: 680511 Rat
    • Entrez Gene: 682330 Rat
    • Omim: 602810 Human
    • SwissProt: P68432 Cow
    • SwissProt: P68431 Human
    • SwissProt: Q6LED0 Rat
    • Unigene: 132854 Human
    • Unigene: 247813 Human
    • Unigene: 247814 Human
    • Unigene: 248176 Human
    • Unigene: 443021 Human
    • Unigene: 484990 Human
    • Unigene: 532144 Human
    • Unigene: 533292 Human
    • Unigene: 546315 Human
    • Unigene: 586261 Human
    • Unigene: 591778 Human
    • Unigene: 138090 Rat
    see all
  • 别名

    • H3 histone family member E pseudogene antibody
    • H3 histone family, member A antibody
    • H3/A antibody
    • H31_HUMAN antibody
    • H3F3 antibody
    • H3FA antibody
    • Hist1h3a antibody
    • HIST1H3B antibody
    • HIST1H3C antibody
    • HIST1H3D antibody
    • HIST1H3E antibody
    • HIST1H3F antibody
    • HIST1H3G antibody
    • HIST1H3H antibody
    • HIST1H3I antibody
    • HIST1H3J antibody
    • HIST3H3 antibody
    • histone 1, H3a antibody
    • Histone cluster 1, H3a antibody
    • Histone H3 3 pseudogene antibody
    • Histone H3.1 antibody
    • Histone H3/a antibody
    • Histone H3/b antibody
    • Histone H3/c antibody
    • Histone H3/d antibody
    • Histone H3/f antibody
    • Histone H3/h antibody
    • Histone H3/i antibody
    • Histone H3/j antibody
    • Histone H3/k antibody
    • Histone H3/l antibody
    see all

图片

  • ChIP - Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)
    ChIP - Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)

    Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The  ChIP was performed with 25 µg of chromatin, 2 µg of  ab2621 (blue), and 20 µl of protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR. Primers and probes are located in the first kb of the transcribed region.

  • Western blot - Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)
    Western blot - Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621)
    All lanes : Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621) at 1 µg/ml

    Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate
    Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (unmodified ) peptide (ab4558) at 0.5 µg/ml
    Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (mono methyl K79) peptide (ab4555) at 0.5 µg/ml
    Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K79) peptide (ab4556) at 0.5 µg/ml
    Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (tri methyl K79) peptide (ab4557) at 0.5 µg/ml
    Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H4 (di methyl K79) peptide (ab4560) at 0.5 µg/ml
    Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate with Human Histone H3 (di methyl K9) peptide (ab1772) at 0.5 µg/ml

    Lysates/proteins at 0.5 µg per lane.

    Secondary
    All lanes : IRDye 680 Conjugated Goat Anti-Rabbit IgG (H+L) at 1/10000 dilution

    Performed under reducing conditions.

    Predicted band size: 15 kDa
    Observed band size: 17 kDa why is the actual band size different from the predicted?

实验方案

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

Click here to view the general protocols

数据表及文件

  • SDS download

  • Datasheet download

    Download

文献 (132)

发表研究结果有使用 ab2621?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab2621 被引用在 132 文献中.

  • Wang SW  et al. Hesperetin promotes DOT1L degradation and reduces histone H3K79 methylation to inhibit gastric cancer metastasis. Phytomedicine 84:153499 (2021). PubMed: 33667841
  • Cao K  et al. DOT1L-controlled cell-fate determination and transcription elongation are independent of H3K79 methylation. Proc Natl Acad Sci U S A 117:27365-27373 (2020). PubMed: 33077595
  • Ryu HY  et al. Histone sumoylation promotes Set3 histone-deacetylase complex-mediated transcriptional regulation. Nucleic Acids Res 48:12151-12168 (2020). PubMed: 33231641
  • Lerner AM  et al. An optogenetic switch for the Set2 methyltransferase provides evidence for transcription-dependent and -independent dynamics of H3K36 methylation. Genome Res 30:1605-1617 (2020). PubMed: 33020206
  • Huang H  et al. Covalent inhibition of NSD1 histone methyltransferase. Nat Chem Biol 16:1403-1410 (2020). PubMed: 32868895
View all Publications for this product

客户评价及客户问答

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1-10 of 15 Abreviews or Q&A

Western blot abreview for Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Fruit fly (Drosophila melanogaster) Tissue lysate - nuclear (whole fruit fly)
Gel Running Conditions
Reduced Denaturing
Loading amount
4.5 µg
Specification
whole fruit fly
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.

Mr. Alexander Hambly

Verified customer

提交于 Apr 14 2015

Immunocytochemistry/ Immunofluorescence abreview for Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Sample
Human Cell (HEK293)
Specification
HEK293
Permeabilization
Yes - TRITON
Fixative
Paraformaldehyde
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 20 2014

ChIP abreview for Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
ChIP
Detection step
Real-time PCR
Sample
Human Cell lysate - whole cell (SCC9 and OKF6)
Specification
SCC9 and OKF6
Negative control
IgG
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 10 minute(s) and 0 second(s)
Specification of the cross-linking agent: formaldehyde
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.

Mr. Dan Stummer

Verified customer

提交于 Apr 11 2014

ChIP abreview for Anti-Histone H3 (methylated) antibody - ChIP Grade

Good
Abreviews
Abreviews
abreview image
Application
ChIP
Sample
Human Cell lysate - nuclear (HEK293)
Specification
HEK293
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 20 minute(s) and 0 second(s)
Specification of the cross-linking agent: 1% Formaldehyde
Detection step
Real-time PCR
Negative control
No antibody
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 Jan 30 2012

Western blot abreview for Anti-Histone H3 (methylated) antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Cell lysate - whole cell (Mouse ES cells)
Loading amount
30 µg
Specification
Mouse ES cells
Gel Running Conditions
Reduced Denaturing (15% SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°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 Nov 07 2011

Western blot abreview for Anti-Histone H3 (methylated) antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Hela Cells)
Loading amount
50 µg
Specification
Hela Cells
Gel Running Conditions
Reduced Denaturing (15% SDS-PAGE)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: RT°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 Nov 07 2011

ChIP abreview for Anti-Histone H3 (methylated) antibody - ChIP Grade

Poor
Abreviews
Abreviews
abreview image
Application
ChIP
Sample
Mouse Cell lysate - whole cell (teratocarcinoma F9 cells)
Specification
teratocarcinoma F9 cells
Type
Cross-linking (X-ChIP)
Duration of cross-linking step: 0 second(s)
Specification of the cross-linking agent: DSG and formaldehyde
Detection step
Real-time PCR
Positive control
H3K4 trimethyl
Negative control
Rabbit IgG
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.

Mr. Dan Stummer

Verified customer

提交于 Mar 15 2011

Western blot abreview for Anti-Histone H3 (methylated) antibody - ChIP Grade

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - nuclear (Liver)
Loading amount
20 µg
Specification
Liver
Treatment
X protein was knockdowned
Gel Running Conditions
Reduced Denaturing (15)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°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 Feb 24 2011

Question

I am interested in purchasing this antibody, but I would like to see the QC data for the current lot.

Read More

Abcam community

Verified customer

Asked on Jan 10 2013

Answer

Here is the WB and ChIP data for lot GR65540-1 (attached) as well as the reviewers notes below. The WB image and ChIP data are consistent with that shown on the datasheet. The only difference is the low signal with ALDOA in ChIP but the stock control run at the same time showed the same low signal so the reviewer wasn’t concerned.
7. WB pass. Lot pass. Pass WB - good blocking with immunising peptide. Can see xr with diK79 peptide but datasheet mentions so allowed. Waiting on ChIP results. SL 22.12.09 Passed in ChIP. The GAPDH signal is variable in these HeLa's and the ALDOA signal is extremely low (even in the STOCK) so this loci is to be ignored in analysis. Passing the ChIP as the test AP is stronger than the STOCK and there is a 2 fold difference between the active and inactive enrichment. DRC 10/01/2010.
WB image caption:



All lanes : Anti-Histone H3 (tri methyl K79) antibody - ChIP Grade (ab2621) at 1 µg/ml

Lane 1 : Calf Thymus Histone Preparation Nuclear Lysate (ab121)
Lane 2 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with http://lab.adminsite.com/Human-Histone-H3-unmodified--peptide-ab4558.html at 0.5 µg/ml
Lane 3 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with http://lab.adminsite.com/Human-Histone-H3-di-methyl-K9-peptide-ab1772.html at 0.5 µg/ml
Lane 4 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with http://lab.adminsite.com/Human-Histone-H3-mono-methyl-K79-peptide-ab4555.html at 0.5 µg/ml
Lane 5 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with http://lab.adminsite.com/Human-Histone-H3-di-methyl-K79-peptide-ab4556.html at 0.5 µg/ml
Lane 6 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with http://lab.adminsite.com/Human-Histone-H3-tri-methyl-K79-peptide-ab4557.html at 0.5 µg/ml
Lane 7 : Calf Thymus Histone Preparation Nuclear Lysate (ab121) with http://lab.adminsite.com/Human-Histone-H4-di-methyl-K79-peptide-ab4560.html at 0.5 µg/ml

Lysates/proteins at 0.5 µg per lane.

Secondary
Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) (ab65484) at 1/3000 dilution

Performed under reducing conditions.

Predicted band size : 15 kDa


Exposure time : 90 seconds

Read More

Abcam Scientific Support

回复于 Jan 10 2013

Question

I want to perform ChIP on Chip experiments. For this purpose I´m planing to use
antibodies from your company which will be verified first in simple ChIP experiments. Our experiences are that the
efficiency of different lots of the same antibody largely varies from lot to lot in ChIP experiments, meaning that verifying the
ChIP grade of an antibody now to perform ChIP on Chip in half a year is meaningless if the lot of the antibody changes in near future.
Thus my question is, is it possible to order antibodies of a certain lot, verify their functionality in my
experimental set up and re-order the antibody with the same lot number in e.g.3-6 months?

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Abcam community

Verified customer

Asked on Jan 23 2012

Answer

We are not able to "save" specific vials of antibody for customer that may wish to purchase them in the future. If you'd like I can recommend specific lots that have the most stock, which may give you the best chance to buy an antibody at a later date from that same stock. The downside with this scenario is that it will only increase the likelihood the specific lot will be available but not guarantee it.

Also, perhaps shortening the time frame for testing the antibodies might also give us a better chance to provide you with the lots you need.

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Abcam Scientific Support

回复于 Jan 23 2012

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