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Cell Biology Cell Cycle Markers

Anti-BrdU抗体[IIB5] (ab8152)

  • Datasheet
  • SDS
Reviews (7)Q&A (5)References (67)

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Immunocytochemistry/ Immunofluorescence - Anti-BrdU antibody [IIB5] (ab8152)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BrdU antibody [IIB5] (ab8152)
  • Flow Cytometry - Anti-BrdU antibody [IIB5] (ab8152)

Key features and details

  • Mouse monoclonal [IIB5] to BrdU
  • Suitable for: IHC-FoFr, ICC/IF, Flow Cyt, IHC-Fr, IHC-P
  • Reacts with: Species independent
  • Isotype: IgG1

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Alexa Fluor® 647 Anti-BrdU antibody [BU1/75 (ICR1)] (ab220075)

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概述

  • 产品名称

    Anti-BrdU抗体[IIB5]
    参阅全部 BrdU 一抗
  • 描述

    小鼠单克隆抗体[IIB5] to BrdU
  • 宿主

    Mouse
  • 特异性

    BrdU is a thymidine analogue and when offered to proliferating cells it is incroporated into reduplicating cells. The antibody is specific for DNA in which BrdU has been incorporated. In immunoassays this antibody reacts strongly with free or carrier-protein coupled BrdU but not with other nucleosides. In immuncytochemistry the antibody only recognizes BrdU in denaturated (single stranded) DNA. The BrdU antibody is 100% crossreactive with Iodo-Deoxy-Uridine (IrdU). Therfore, IdU instead of BrdU can be used in studies.
  • 经测试应用

    适用于: IHC-FoFr, ICC/IF, Flow Cyt, IHC-Fr, IHC-Pmore details
  • 种属反应性

    与反应: Species independent
  • 免疫原

    Chemical/ Small Molecule corresponding to BrdU conjugated to bovine serum albumin.

  • 阳性对照

    • Bromodeoxyuridine labeled cells.
  • 常规说明

    The following product is available as purified antibody (purified by affinity chromatography) together with several conjugated versions:

    Anti-BrdU antibody [BU1/75 (ICR1)] (ab6326)

    Unstained positive control slides from mice treated with BrdU (formalin-fixed, paraffin-embedded intestine sections) are available as BrdU control slides ab129956.

     

    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 long term.
  • 存储溶液

    pH: 7.3
    Preservative: 0.09% Sodium azide
    Constituents: 1% BSA, PBS
  • Concentration information loading...
  • 纯度

    Tissue culture supernatant
  • 克隆

    单克隆
  • 克隆编号

    IIB5
  • 骨髓瘤

    Sp2/0-Ag14
  • 同种型

    IgG1
  • 研究领域

    • Cell Biology
    • Cell Cycle
    • Markers
    • Neuroscience
    • Cell Type Marker
    • Neuron marker
    • Soma marker
    • Tags & Cell Markers
    • Cell Type Markers
    • Replication
    • Epigenetics and Nuclear Signaling
    • DNA / RNA
    • DNA / Nucleotides

相关产品

  • Alternative Versions

    • Anti-BrdU antibody [IIB5] (ab8955)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
  • Related Buffer

    • BrdU (5-bromo-2'-deoxyuridine), Thymidine analog (ab142567)
  • Related Products

    • Anti-BrdU antibody [BU1/75 (ICR1)] - Proliferation Marker (ab6326)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab8152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
IHC-FoFr (1)
Use at an assay dependent concentration.
ICC/IF (4)
Use at an assay dependent concentration.
Flow Cyt
1/100 - 1/200.

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

IHC-Fr
1/5 - 1/10.

Dilute antibody in 0.15 M phosphate buffered saline with 1% BSA and 1% Na-azide.

IHC-P (2)
1/5 - 1/10. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Enzymatic antigen retrieval with proteases can also be used.

说明
IHC-FoFr
Use at an assay dependent concentration.
ICC/IF
Use at an assay dependent concentration.
Flow Cyt
1/100 - 1/200.

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

IHC-Fr
1/5 - 1/10.

Dilute antibody in 0.15 M phosphate buffered saline with 1% BSA and 1% Na-azide.

IHC-P
1/5 - 1/10. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

Enzymatic antigen retrieval with proteases can also be used.

靶标

  • 相关性

    The immunocytochemical detection of bromodeoxyuridine (BrdU) incorporated into DNA is a powerful tool to study the cytokinetics of normal and neoplastic cells. In vitro or in vivo labeling of tumor cells with the thymidine analogue BrdU and the subsequent detection of incorporated BrdU with specific anti-BrdU monoclonal antibodies is an accurate and comprehensive method to quantitate the degree of DNA-synthesis. BrdU is incorporated into the newly synthezised DNA of S-phase cells may provide an estimate for the fraction of cells in S-phase. Also dynamic proliferative information such as the S-phase transit rate and the potential doubling time can be obtained, by means of bivariate BrdU/DNA flow cytometric analysis.
  • 细胞定位

    Nuclear
  • 别名

    • Bromodeoxyuridine antibody
    • BUdr antibody

图片

  • Immunocytochemistry/ Immunofluorescence - Anti-BrdU antibody [IIB5] (ab8152)
    Immunocytochemistry/ Immunofluorescence - Anti-BrdU antibody [IIB5] (ab8152)Image courtesy of an Abreview submitted by Dr. Kirk McManus, Univ. of Manitoba/Cancer Care MICB, Canada
    ab8152 (1/100) staining BrdU in HeLa cells (green). Cells were fixed with paraformaldehyde, permeabilized with 0.5% Triton X100/ PBS and counterstained with DAPI in order to highlight the nucleus (red). For further esperimental details please see Abreview.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BrdU antibody [IIB5] (ab8152)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BrdU antibody [IIB5] (ab8152)This image is courtesy of an anonymous Abreview
    Immunohistochemical analysis of rat brain tissue, staining BrdU with ab8152.

    Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at 25°C; antigen retrieval was by heat mediation in citrate buffer (pH 6). Samples were incubated with primary antibody (1/20 in diluent) for 18 hours at 25°C. A Cy3®-conjugated donkey anti-mouse polyclonal IgG was used as the secondary antibody.

    See Abreview

  • Flow Cytometry - Anti-BrdU antibody [IIB5] (ab8152)
    Flow Cytometry - Anti-BrdU antibody [IIB5] (ab8152)

    Cells were pulse labeled with 10 µM BrdU for 30 min, rinsed twice in prewarmed PBS, and chased in prewarmed culture medium, supplemented with 5 mM deoxythymidine. Incorporated BrdU was detected after ethanol fixation of the cells, which were than rinsed once in PBS and resuspended in 2 ml of 0.4 mg/ml pepsin in 0.1 N HCl. After 30 min at room temperature cells were pelleted, resuspended in 2 N HCl, and incubated for another 30 min at 37°C. Cells were rinsed in 0.1 M borate buffer, pH 8.5, and PBS/BSA (1 mg/ml BSA in PBS). Appropriately diluted mouse anti-BrdU antibody (clone IIB5) was added to the cell pellet, resuspended in 100 micro liters PBS/BSA. After incubation for 1 h at room temperature, the cells were rinsed twice in PBS/BSA. For visualization, FITC-conjugated Fab2 fragments of rabbit anti-mouse Ig antibody were added in a 1/10 dilution. After incubation for 45 min at room temperature samples were rinsed twice in PBS/BSA and the cells were finally resuspended in 0.5 ml cold PBS supplemented with 100 microgram/ml RNAse and 20 µg/mL propidium iodide. The samples were allowed to stand for 15 min on ice in the dark before flow cytometric analysis. In the negative control the primary antibody was omitted.

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 (67)

发表研究结果有使用 ab8152?请让我们知道,以便我们可以引用本数据表中的参考文章。

ab8152 被引用在 67 文献中.

  • Dubois-Pot-Schneider H  et al. Transcriptional and Epigenetic Consequences of DMSO Treatment on HepaRG Cells. Cells 11:N/A (2022). PubMed: 35892596
  • Ghosh B  et al. Loss of E-cadherin is causal to pathologic changes in chronic lung disease. Commun Biol 5:1149 (2022). PubMed: 36309587
  • Xu S  et al. Leukemia inhibitory factor is a therapeutic target for renal interstitial fibrosis. EBioMedicine 86:104312 (2022). PubMed: 36335669
  • Vlach M  et al. Liposome-Mediated Gene Transfer in Differentiated HepaRG™ Cells: Expression of Liver Specific Functions and Application to the Cytochrome P450 2D6 Expression. Cells 11:N/A (2022). PubMed: 36497165
  • Hardy RA  et al. Role of age and neuroinflammation in the mechanism of cognitive deficits in sickle cell disease. Exp Biol Med (Maywood) 246:106-120 (2021). PubMed: 32962408
View all Publications for this product

客户评价及客户问答

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提交评价 提交问题

1-10 of 12 Abreviews or Q&A

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-BrdU antibody [IIB5]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Rabbit Tissue sections (Intestine)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: 2M HCl
Permeabilization
Yes - Triton X-100
Specification
Intestine
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
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.

DR. Egi Kardia

Verified customer

提交于 Sep 21 2018

Immunocytochemistry/ Immunofluorescence abreview for Anti-BrdU antibody [IIB5]

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rabbit Cell (Primary airway epithelial cells)
Permeabilization
No
Specification
Primary airway epithelial cells
Blocking step
BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: RT°C
Fixative
70% Ethanol
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.

DR. Egi Kardia

Verified customer

提交于 Sep 06 2018

Immunocytochemistry/ Immunofluorescence abreview for Anti-BrdU antibody [IIB5]

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Blocking step
BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 22°C
Sample
Human Cell (OCI ly1)
Specification
OCI ly1
Permeabilization
Yes - 0.25% Triton X-100 in PBS
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 Nov 07 2014

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-BrdU [IIB5] antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (brain)
Specification
brain
Fixative
Paraformaldehyde
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization
No
Blocking step
Serum as blocking agent for 30 minute(s) · Concentration: 10% · 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

提交于 Dec 07 2012

Immunocytochemistry/ Immunofluorescence abreview for Anti-BrdU [IIB5] antibody

Excellent
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
African Green Monkey Cell (Oligodendrocytes, astrocytes (mixed glial culture))
Specification
Oligodendrocytes, astrocytes (mixed glial culture)
Fixative
Paraformaldehyde
Permeabilization
Yes - 0.3% Triton-PBS for 30 minutes followed by DNAse treatment for 90 minutes.
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 28 2010

Question

Hello, I need to work with IdU and CldU for double staining. I found out that I can use for cldU the ab6326 that doen't cross react with IdU. Can I use ab8039 or ab8152 for IdU? I need an antibody that doesn't cross react with CldU.

Read More

Abcam community

Verified customer

Asked on Apr 03 2013

Answer

Thank you for your inquiry and for rating your experience with us.

I just wanted to follow up on your query with some additional information.

For ab8039, this antibody can be used for detection of BrdU and BrU. We have not tested it regarding other chemicals of this family.

For ab8152, this antibody is specific for DNA in which BrdU has been incorporated. In immunoassays this antibody reacts strongly with free or carrier-protein coupled BrdU but not with other nucleosides. In immuncytochemistry the antibody only recognizes BrdU in denaturated (single stranded) DNA. The BrdU antibody is 100% crossreactive with Iodo-Deoxy-Uridine (IrdU). Therfore, IdU instead of BrdU can be used in studies.

I hope this information helps. Please contact us with any other questions.

Read More

Abcam Scientific Support

回复于 Apr 03 2013

Question

the floating sections are not frozen.all steps are performed with the sections positioned in12 well plates.

it is not possible to work withthem on slides because they immediately come off . once they are immersed in citric buffer they will surely come off.

i ordered the HIER solution you offered , but i wonder how to work with it in the conditions specified above.

Read More

Abcam community

Verified customer

Asked on Jun 19 2012

Answer

Thank you for contacting us.

For floating sections, please check thefollowing protocol on IHC World website:
http://www.ihcworld.com/_protocols/epitope_retrieval/free_floating_sections.htm
This protocol is for paraffin-embedded as well as frozen sections.

As for ensuring the paraffin sections stick to your slides:
Different sections and tissues stick with differetn strength to the slides.
It might be advisable to use charged slides, as well as to bake your sections onto the slides by exposing them to heat (e.g. heating plate or oven) up to 60 degree Celsius.The purpose is to remove miscroscopic amounts of water between the sections and the slide.
Subsequently the paraffin would need to removed according to a standard IHC-P protocol. Then you would proceed with the antigen retrieval step.

Also, please check this information on the IHC World website for further advice.
http://www.ihcworld.com/_faq/histology-faq/section/s1.htm

Read More

Abcam Scientific Support

回复于 Jun 19 2012

Question

how do i technically imerse the tissue sedctions in the citrate buffer without loosing them?

if sections are on a slide, and the slide is imersed in the boiling buffer, the sections will detach from the slides.

also, i am working with floating sections. do ypu have a technical tip re howthis step should be excecuted?

Read More

Abcam community

Verified customer

Asked on Jun 14 2012

Answer

The slides need to be immersed, for the retrieval to be effective. Yes, the sections could come off ifthey are kept too long in the retrieval solution. Should they come off, you would need to reduce the time they are exposed to the retrieval buffer.

Are the floating sections paraffin-embedded or frozen? I have found the following protocol on IHC World website which should help you further:
http://www.ihcworld.com/_protocols/epitope_retrieval/free_floating_sections.htm

Read More

Abcam Scientific Support

回复于 Jun 14 2012

Question

Inquiry: what type of antigen retreival is required in order to detect the brdu in PFA sixed tissue sections, along with doublecortin detection? the protocol that uses HCl does not allow dapi counterstain.and also destroys the other antigen ,which is cytosolic. if citric acid should be used- can you send me the excat protocol? including steps of trypsin, etc. which secondary Ab should be used and at what concentration?

Read More

Abcam community

Verified customer

Asked on Jun 05 2012

Answer

I have checked with the lab:
Antigen retrieval can be done with HIER using citrate buffer.

Please see below for the protocol:

High Temperature antigen Unmasking Technique using Sodium Citrate Buffer for Immunohistochemical Demonstration on Paraffin Sections

1. Cut and mount sections on slides coated with Vectabond or APES (3-aminopropyltriethoxysilane).
2. Deparaffinize sections and rehydrate to distilled water.
3. Bring 1600ml 0.01M sodium citrate buffer (pH 6.0) to the boil in a Prestige stainless steel pressure cooker, using a hot plate. Cover but do not lock lid.
4. Position slides into metal staining racks and lower into pressure cooker ensuring slides are well immersed in citrate buffer. Lock lid. The small valve will rise.
5. When the pressure indicator valve (the large one) has risen after about 4 minutes, incubate sections for 1 minute.
6. Remove pressure cooker from heat source and run under cold water with lid on. When the small valve sinks open lid and remove slides and place immediately into distilled water. DO NOT OPEN LID UNTIL THE SMALL VALVE SINKS.
7. Wash sections in TBS buffer (pH 7.6) for 1 x 5 minutes.
8. Place sections in 1.5% hydrogen peroxide/methanol for 10 minutes.
9. Wash sections in distilled water for 2 x 5 minutes, then wash sections in TBS buffer for 2 x 5 minutes.
10. Place sections in normal serum for 20 minutes.
11. Cover sections with primary antibody. ( The optimal dilution of the antibody, incubation time and incubation temperature should be determined by the individual laboratory).
12. Wash in TBS buffer for 2 x 5 minutes.
13. Incubate sections in secondary antibody for 30 minutes.
14. Wash in TBS buffer for 2 x 5 minutes.
15. Incubate slides in ABComplex for 30 minutes.
16. Wash in TBS buffer for 2 x 5 minutes.
17. Incubate slides in DAB.
18. Wash in water for 2 x 5 minutes.
19. Counterstain with haematoxylin (if required), dehydrate, coverslip and mount.


To avoid sections becoming detached, sections should be mounted on Vectabond or APES covered slides, then dried at 37oC overnight followed by drying at 56oC for 60 minutes.

Read More

Abcam Scientific Support

回复于 Jun 05 2012

Question

Do you know how long should be the ssDNA to be detected with the ab by Immunofluorescence? I want to use is to detect fragment between 30-300 nt fron adherent cells in culture.

Read More

Abcam community

Verified customer

Asked on Nov 18 2011

Answer

Thank you for contacting us with your question. We unfortunately do not have any data regarding the minimum length of ssDNA required for antibody staining on adherent cells.

Read More

Abcam Scientific Support

回复于 Nov 18 2011

1-10 of 12 Abreviews or Q&A

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