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Epigenetics and Nuclear Signaling Transcription Domain Families Zinc Finger
重组RabMAb

重组Anti-Progesterone Receptor抗体[YR85] (ab32085)

  • Datasheet
  • SDS
Reviews (5)Q&A (4)References (35)

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Immunocytochemistry/ Immunofluorescence - Anti-Progesterone Receptor antibody [YR85] (ab32085)
  • Immunocytochemistry/ Immunofluorescence - Anti-Progesterone Receptor antibody [YR85] (ab32085)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Progesterone Receptor antibody [YR85] (ab32085)
  • Flow Cytometry (Intracellular) - Anti-Progesterone Receptor antibody [YR85] (ab32085)
  • Western blot - Anti-Progesterone Receptor antibody [YR85] (ab32085)
  • Western blot - Anti-Progesterone Receptor antibody [YR85] (ab32085)
  • Immunocytochemistry/ Immunofluorescence - Anti-Progesterone Receptor antibody [YR85] (ab32085)
  • OI-RD Scanning - Anti-Progesterone Receptor antibody [YR85] (ab32085)
  • Anti-Progesterone Receptor antibody [YR85] (ab32085)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [YR85] to Progesterone Receptor
  • Suitable for: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IF
  • Reacts with: Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 568 Alexa Fluor® 594 Alexa Fluor® 647 Alkaline Phosphatase APC Carrier Free HRP PE

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概述

  • 产品名称

    Anti-Progesterone Receptor抗体[YR85]
    参阅全部 Progesterone Receptor 一抗
  • 描述

    兔单克隆抗体[YR85] to Progesterone Receptor
  • 宿主

    Rabbit
  • 特异性

    This antibody recognises progesterone receptor. The antibody does not cross-react with other NR3 family members. Since the recognized epitope is near the N-terminal end of the protein, this product should only detect isoform B and not isoform A.

  • 经测试应用

    适用于: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IFmore details
  • 种属反应性

    与反应: Human
  • 免疫原

    Synthetic peptide within Human Progesterone Receptor (N terminal). The exact sequence is proprietary.
    Database link: P06401

  • 阳性对照

    • Human breast carcinoma, T47D cell lysate. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: DU145.
  • 常规说明

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    Mouse, Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

性能

  • 形式

    Liquid
  • 存放说明

    Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
  • 解离常数(KD)

    KD = 2.48 x 10 -10 M
    Learn more about KD
  • 存储溶液

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 49% PBS, 50% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...
  • 纯度

    Protein A purified
  • 克隆

    单克隆
  • 克隆编号

    YR85
  • 同种型

    IgG
  • 研究领域

    • Epigenetics and Nuclear Signaling
    • Transcription
    • Domain Families
    • Zinc Finger
    • Signal Transduction
    • Signaling Pathway
    • Nuclear Signaling
    • Nuclear Hormone Receptors
    • Progesterone
    • Neuroscience
    • Endocrine system
    • Gonadotrophic axis
    • Epigenetics and Nuclear Signaling
    • Nuclear Signaling Pathways
    • Nuclear Receptors
    • Progesterone
    • Cancer
    • Signal transduction
    • Nuclear signaling
    • Nuclear hormone receptors
    • Progesterone
    • Cancer
    • Tumor biomarkers
    • Receptors
    • Neuroscience
    • Development
    • Neuroscience
    • Processes

相关产品

  • Alternative Versions

    • Anti-Progesterone Receptor antibody [YR85] - BSA and Azide free (ab206926)
    • Alexa Fluor® 568 Anti-Progesterone Receptor antibody [YR85] (ab312430)
  • Compatible Secondaries

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

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • T-47D whole cell lysate (ab14899)
  • Recombinant Protein

    • Recombinant Human Progesterone Receptor protein (ab159105)

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab32085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 (Intra)
Use at an assay dependent concentration.
WB
1/1000 - 1/10000. Predicted molecular weight: 99 kDa.
IHC-P (4)
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
1/150.
ICC/IF
Use a concentration of 0.2 - 5 µg/ml.
说明
Flow Cyt (Intra)
Use at an assay dependent concentration.
WB
1/1000 - 1/10000. Predicted molecular weight: 99 kDa.
IHC-P
1/100 - 1/250. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
IP
1/150.
ICC/IF
Use a concentration of 0.2 - 5 µg/ml.

靶标

  • 功能

    The steroid hormones and their receptors are involved in the regulation of eukaryotic gene expression and affect cellular proliferation and differentiation in target tissues. Progesterone receptor isoform B (PRB) is involved activation of c-SRC/MAPK signaling on hormone stimulation.
    Isoform A: inactive in stimulating c-Src/MAPK signaling on hormone stimulation.
    Isoform 4: Increases mitochondrial membrane potential and cellular respiration upon stimulation by progesterone.
  • 序列相似性

    Belongs to the nuclear hormone receptor family. NR3 subfamily.
    Contains 1 nuclear receptor DNA-binding domain.
  • 结构域

    Composed of three domains: a modulating N-terminal domain, a DNA-binding domain and a C-terminal ligand-binding domain.
  • 翻译后修饰

    Phosphorylated on multiple serine sites. Several of these sites are hormone-dependent. Phosphorylation on Ser-294 occurs preferentially on isoform B, is highly hormone-dependent and modulates ubiquitination and sumoylation on Lys-388. Phosphorylation on Ser-102 and Ser-345 also requires induction by hormone. Basal phosphorylation on Ser-81, Ser-162, Ser-190 and Ser-400 is increased in response to progesterone and can be phosphorylated in vitro by the CDK2-A1 complex. Increased levels of phosphorylation on Ser-400 also in the presence of EGF, heregulin, IGF, PMA and FBS. Phosphorylation at this site by CDK2 is ligand-independent, and increases nuclear translocation and transcriptional activity. Phosphorylation at Ser-162 and Ser-294, but not at Ser-190, is impaired during the G(2)/M phase of the cell cycle. Phosphorylation on Ser-345 by ERK1/2 MAPK is required for interaction with SP1.
    Sumoylation is hormone-dependent and represses transcriptional activity. Sumoylation on all three sites is enhanced by PIAS3. Desumoylated by SENP1. Sumoylation on Lys-388, the main site of sumoylation, is repressed by ubiquitination on the same site, and modulated by phosphorylation at Ser-294.
    Ubiquitination is hormone-dependent and represses sumoylation on the same site. Promoted by MAPK-mediated phosphorylation on Ser-294.
    Palmitoylated by ZDHHC7 and ZDHHC21. Palmitoylation is required for plasma membrane targeting and for rapid intracellular signaling via ERK and AKT kinases and cAMP generation.
  • 细胞定位

    Nucleus. Cytoplasm. Nucleoplasmic shuttling is both homone- and cell cycle-dependent. On hormone stimulation, retained in the cytoplasm in the G(1) and G(2)/M phases; Mitochondrion outer membrane and Nucleus. Cytoplasm. Mainly nuclear.
  • Target information above from: UniProt accession P06401 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: 5241 Human
    • Omim: 607311 Human
    • SwissProt: P06401 Human
    • Unigene: 32405 Human
    • Unigene: 742403 Human
    • 别名

      • NR3C3 antibody
      • Nuclear receptor subfamily 3 group C member 3 antibody
      • PGR antibody
      • PR antibody
      • PRA antibody
      • PRB antibody
      • PRGR_HUMAN antibody
      • Progesterone receptor antibody
      • Progestin receptor form A antibody
      • Progestin receptor form B antibody
      see all

    图片

    • Immunocytochemistry/ Immunofluorescence - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Immunocytochemistry/ Immunofluorescence - Anti-Progesterone Receptor antibody [YR85] (ab32085)

      Immunofluorescence staining of Recombinant Anti-Progesterone Receptor antibody [YR85] (ab32085) in T47D cells (+ve control) and HepG2 cells (-ve control). The cells were fixed with 100% Methanol (5 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab32085 at 0.2 µg/mL dilution and ab7291, Anti-alpha Tubulin antibody [DM1A], at 1/1000 dilution, followed by a further incubation at room temperature for 1h with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150087) at 1/1000 (shown in red) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 1/1000 (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with the Perkin Elmer Operetta and a maximum intensity projection of confocal planes is shown.

    • Immunocytochemistry/ Immunofluorescence - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Immunocytochemistry/ Immunofluorescence - Anti-Progesterone Receptor antibody [YR85] (ab32085)

      Immunofluorescence staining of Recombinant Anti-Progesterone Receptor antibody [YR85] (ab32085) in T47D cells (+ve control) and HepG2 cells (-ve control). The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab32085 at 0.2 µg/mL dilution and ab7291, Anti-alpha Tubulin antibody [DM1A], at 1/1000 dilution, followed by a further incubation at room temperature for 1h with Goat Anti-Rabbit IgG H&L (Alexa Fluor® 647) preadsorbed (ab150087) at 1/1000 (shown in red) and Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 1/1000 (shown in green). Nuclear DNA was labelled with DAPI (shown in blue). Image was taken with the Perkin Elmer Operetta and a maximum intensity projection of confocal planes is shown.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Progesterone Receptor antibody [YR85] (ab32085)

      Immunohistochemical analysis of progesterone receptor expression in paraffin embedded human breast carcinoma, using 1/100 ab32085.

      Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

    • Flow Cytometry (Intracellular) - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Flow Cytometry (Intracellular) - Anti-Progesterone Receptor antibody [YR85] (ab32085)

      Intracellular Flow Cytometry analysis of T47D (human mammary gland ductal carcinoma) cells labeling Progesterone Receptor with purified ab32085 at 1/1100 dilution (1ug/ml) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488) (ab150077) (1/2000 dilution) was used as the secondary antibody. Rabbit monoclonal IgG (Black) (ab172730) was used as the isotype control, Cell without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control. 

       

       

    • Western blot - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Western blot - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Anti-Progesterone Receptor antibody [YR85] (ab32085) at 1/1000 dilution + Human breast cancer lysate at 15 µg

      Secondary
      Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution

      Predicted band size: 99 kDa
      Observed band size: 118 kDa why is the actual band size different from the predicted?


      Exposure time: 180 seconds
    • Western blot - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Western blot - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Anti-Progesterone Receptor antibody [YR85] (ab32085) at 1/2000 dilution + T-47D (Human ductal breast epithelial tumor epithelial cell) whole cell lysate at 15 µg

      Secondary
      Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 99 kDa
      Observed band size: 118 kDa why is the actual band size different from the predicted?


      Exposure time: 40 seconds


      Blocking buffer: 5% NFDM/TBST.

    • Immunocytochemistry/ Immunofluorescence - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Immunocytochemistry/ Immunofluorescence - Anti-Progesterone Receptor antibody [YR85] (ab32085)

      ICC/IF image of ab32085 stained DU145 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 ab32085 at 5µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- rabbit (ab96899) IgG (H+L) used at a 1/250 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.

    • OI-RD Scanning - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      OI-RD Scanning - Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Equilibrium disassociation constant (KD)
      Learn more about KD

      Click here to learn more about KD
    • Anti-Progesterone Receptor antibody [YR85] (ab32085)
      Anti-Progesterone Receptor antibody [YR85] (ab32085)

    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 (35)

    发表研究结果有使用 ab32085?请让我们知道,以便我们可以引用本数据表中的参考文章。

    ab32085 被引用在 35 文献中.

    • Zhang L  et al. Progesterone receptor distribution in the human hypothalamus and its association with suicide. Acta Neuropathol Commun 12:16 (2024). PubMed: 38263257
    • Wang L  et al. RNA-seq and ATAC-seq analysis of CD163+ macrophage-induced progestin-insensitive endometrial cancer cells. Cancer Med 12:5964-5978 (2023). PubMed: 36373483
    • Lou Y  et al. Expression and clinical significance of undifferentiated embryonic cell transcription factor 1 in breast cancer. Transl Cancer Res 12:150-162 (2023). PubMed: 36760370
    • Du Z  et al. Purine Intake and All-Cause Mortality in Ovarian Cancer: Results from a Prospective Cohort Study. Nutrients 15:N/A (2023). PubMed: 36839289
    • Luo M  et al. E. coli Nissle 1917 ameliorates mitochondrial injury of granulosa cells in polycystic ovary syndrome through promoting gut immune factor IL-22 via gut microbiota and microbial metabolism. Front Immunol 14:1137089 (2023). PubMed: 37275915
    View all Publications for this product

    客户评价及客户问答

    Show All 评价 Q&A
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    1-9 of 9 Abreviews or Q&A

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Progesterone Receptor antibody [YR85]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rhesus monkey Tissue sections (Uterus)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate ph6.0
    Permeabilization
    No
    Specification
    Uterus
    Blocking step
    Serum as blocking agent for 20 minute(s) · Concentration: 2% · Temperature: 23°C
    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 Sep 27 2018

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Progesterone Receptor antibody [YR85]

    Poor
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Pig Tissue sections (Mammary epithelial cells)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 10mM citrate buffer
    Permeabilization
    No
    Specification
    Mammary epithelial cells
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 23°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.

    Mrs. Katie Elliott

    Verified customer

    提交于 Jul 15 2016

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Progesterone Receptor antibody [Y85]

    Excellent
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (human breast cancer xenograft (MCF7))
    Specification
    human breast cancer xenograft (MCF7)
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citrate buffer, pH6.0
    Permeabilization
    No
    Blocking step
    BSA as blocking agent for 20 minute(s) · Concentration: 1% · 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 Apr 26 2010

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-Progesterone Receptor antibody [Y85]

    Good
    Abreviews
    Abreviews
    abreview image
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (breast cancer)
    Specification
    breast cancer
    Fixative
    Formaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: citrate - 20 minutes 100oC
    Permeabilization
    No
    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 09 2009

    ChIP abreview for Anti-Progesterone Receptor antibody [Y85]

    Average
    Abreviews
    Abreviews
    abreview image
    Application
    ChIP
    Sample
    Human Cell lysate - whole cell (Uterus)
    Specification
    Uterus
    Type
    Cross-linking (X-ChIP)
    Duration of cross-linking step: 10 minute(s) and 0 second(s)
    Specification of the cross-linking agent: Formaldehyde
    Detection step
    Semiquantitative PCR
    Positive control
    RNA Polymerase II
    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.

    Abcam user community

    Verified customer

    提交于 Oct 17 2008

    Question



    Inquiry: To the technical staff. Regarding: Problem with product/complaint To nwhom it may concern. For about 1/2 year we have put immens effort into tweaking our methods to get the above antibody to work in rat tissue (bought it because data sheet did say it works in that species), only to find out now, that Abcam simply changed the cross reactivity label to "Reacts with Human, Does not react with Mouse, Rat". I am heavily disappointed, to say the least, as are my coworkers and students, how worked hard on the project. As we only got weak readings (however specific as we believe) we wonder under what circumstances Abcam changed the crossreactivity. This would help us understand our findings better. Moreover, I hope this message helps improve your customer care. As a principle investigator I order antibodies from Abcam on a regular basis. However this is the first time I am deeply dissapointed not beeing informed about such important piece of information.

    Read More

    Abcam community

    Verified customer

    Asked on Jun 08 2012

    Answer

    Es tut mir leid, dass der Antikörper nicht so funktioniert hat wie auf dem Datenblatt angegeben. Ich habe unsere Finanzabteilung beauftragt, Gutschriften für Sie auszustellen. Sie können die Gutschriften auf folgende Weise verwenden:

    (1) Gegen die Rechnung dieses Antikörpers verwenden falls diese noch nicht bezahlt wurde
    (2) Aufbewahren und gegen eine zukünftige Rechnung verwenden
    (3) Wir stellen Ihnen gerne eine komplette Rückerstattung aus, wenn keine offenen Rechnungen mehr ausstehen

    Um eine Rückerstattung des Kaufpreises zu erhalten oder um zu bestätigen wie Sie diese Gutschrift benutzen möchten, möchte ich Sie bitten, sich mit unserer Finanzabteilung in Verbindung zu setzen.
    Sie können unsere Finanzabteilung per Email unter der Adresse creditcontrol@abcam.com oder telefonisch unter der Nummer 030 896 779 154 erreichen.

    Bitte geben Sie die Referenznummer Ihrer Gutschrift immer mit an.

    Die Referenznummer ist für Ihre Unterlagen bestimmt. Wir schicken Ihnen die eigentliche Gutschrift per Post zu. Sie enthält auch die Gutschriftnummer, die mit dem Buchstaben CGB beginnt.

    Ich hoffe dass Sie trotz dieser schlechten Erfahrung weiterhin Kunde bei uns bleiben. Bitte zögern Sie nicht sich wieder an uns zu wenden, falls Sie weitere Fragen haben.

    Read More

    Abcam Scientific Support

    回复于 Jun 08 2012

    Question

    vielen Dank für die Bearbeitung!
    Ich werde Ihnen die Daten umgehend zukommen lassen!
    Meine Arbeitsgruppe interessiert sich natürlich zu erfahren warum der Antikörper nicht mehr für Messungen an der Ratte bestimmt ist, zumal sich hin und wieder positive Färbungen erzielen ließen.

    Read More

    Abcam community

    Verified customer

    Asked on May 31 2012

    Answer

    Wir testen unsere Chargen immer, bevor wir unser Datenblatt aktualisieren, und uns ist bei diesem Antikörper aufgefallen, dass die Spezies- Reaktivität in Maus und Ratte eben nicht konstant, bzw gar nicht mehr gegeben war, so dass wir Maus und Ratte als "reagiert nicht" deklariert haben. Seitdem testen wir die beiden Spezies übrigens auch gar nicht mehr.
    Warum dies selbst bei einem monoklonalem (!) Antikörper passieren kann, ist uns leider auch nicht bekannt.

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    Abcam Scientific Support

    回复于 May 31 2012

    Question

    die Bestellnummer des Antikörpers ist


    ab32085 Rabbit monoclonal [Y85] to Progesterone Receptor

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    Abcam community

    Verified customer

    Asked on May 30 2012

    Answer

    Vielen Dank für Ihre Email.

    Wir haben das Datenblatt im März 2011 geändert, allerdings habe ich keine Hinweise darauf gefunden, dass wir eine Kampagne gestartet haben. Ich habe dies jetzt nachgeholt.

    Wenn Sie mir bitte mitteilen, wann Sie denn Antikörper gekauft haben (ungefähres Datum reicht), kann ich ihnen schon jetzt entweder eine Gutschrift, eine Erstattung oder einen kostenlosen Erstsatz zukommen lassen.

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    Abcam Scientific Support

    回复于 May 30 2012

    Question

    BATCH NUMBER 298365 ORDER NUMBER 257478 DESCRIPTION OF THE PROBLEM No staining SAMPLE kidney PRIMARY ANTIBODY 1:100 in 10% normal goat serum,overnight Wash 3X5min with PBS DETECTION METHOD DAB develope 10min POSITIVE AND NEGATIVE CONTROLS USED ovary ANTIBODY STORAGE CONDITIONS -20C FIXATION OF SAMPLE histochoice ANTIGEN RETRIEVAL Microwave, 0.1 M citrate buffer BLOCKING CONDITIONS 6% H2O2 for 10min, wash 3tX5 min with PBS Avidin 15min, Wash 3X5min with PBS Biotin 15min, Wash 3X5min with PBS 10% normal goat serum 20min SECONDARY ANTIBODY rabbit 1:100 for 1 hour at RT Wash 3X5min with PBS ABC 1 hour at RT Wash 3X5min with PBS HOW MANY TIMES HAVE YOU TRIED THE APPLICATION? 4 HAVE YOU RUN A "NO PRIMARY" CONTROL? Yes DO YOU OBTAIN THE SAME RESULTS EVERY TIME? Yes WHAT STEPS HAVE YOU ALTERED? antigen retrieval or not

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    Abcam community

    Verified customer

    Asked on Sep 11 2007

    Answer

    Thank you for your enquiry. I am sorry to hear that you are experiencing difficulties with this product ab32085 in immunohistochemistry. 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 at the protocols you used and have a few questions and suggestions that might help you determine the cause of the no-staining results. I would therefore appreciate if you can please clarify the following items. - are your kidney samples taken from human? - can you confirm that the second antibody you used also managed to produce good results with other primary antibodies? Was the secondary antibody biotin conjugated? In some cases, the problem may be with the secondary antibody not working. - was the deparaffinization process complete? Did you used a new batch of xylene? - how long was the heating in citrate buffer done? We would normally recommend using 0.01M Citrate Buffer, pH6.0 and heat for 10 min. - After making the ABC complex, how long did you leave it to stand? In general, it will require a minimum of 30 minutes as this is the length of time that the complex takes to form. Did this ABC kit also work well in other recent experiments? I hope the above recommendations may already help you. If you have already tried the above suggestions and still experience problems, please do not hesitate to contact me with your shipping address/purchasing agent information. Also please advice me on how you would like to proceed with your enquiry, so that I can immediately arrange for a replacement or refund to you.

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    Abcam Scientific Support

    回复于 Sep 11 2007

    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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