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SimpleStep

JNK 1/2 (pT183/Y185 + Total) ELISA试剂盒(ab176662)

Reviews (2) Submit a question References (12)
Typical cell lysate dilution series
  • Typical recombinant protein standard curve
  • Linearity of dilution
  • Comparison of JNK1/2 expression in different cell lines
  • JNK1/2 (pT183/Y185) phosphorylation in response to anisomycin treatment
  • Sandwich ELISA - JNK 1/2 (pT183/Y185 + Total) ELISA Kit (ab176662)
  • Sandwich ELISA - JNK 1/2 (pT183/Y185 + Total) ELISA Kit (ab176662)

Key features and details

  • One-wash 90 minute protocol
  • Sample type: Cell Lysate, Tissue Homogenate
  • Detection method: Colorimetric
  • Assay type: Semi-quantitative
  • Reacts with: Mouse, Human

概述

  • 产品名称

    JNK 1/2 (pT183/Y185 + Total) ELISA试剂盒

  • 检测方法

    Colorimetric

  • 精确度

    批次内
    样品 n Mean SD CV%
    (pT183/Y185) 6 2.7%
    (Total) 6 2.8%
    批次间
    样品 n Mean SD CV%
    (pT183/Y185) 3 3.8%
    (Total) 3 3.3%

  • 样品类型

    Cell Lysate, Tissue Homogenate

  • 检测类型

    Semi-quantitative

  • 检测时间

    1h 30m

  • 实验步骤

    One step assay

  • 种属反应性

    与反应: Mouse, Human
    预测可用于: Rat

  • 产品概述

    Abcam’s JNK1/2 (pT183/Y185) and JNK1/2 (Total) in vitro SimpleStep ELISA™ (Enzyme-Linked Immunosorbent Assay) kit is designed for the semi-quantitative measurement of JNK1/2 (pT183/Y185) and Total JNK1/2 protein in Human and mouse cells.


    The SimpleStep ELISA™ employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. TMB substrate is added and during incubation is catalyzed by HRP, generating blue coloration. This reaction is then stopped by addition of Stop Solution completing any color change from blue to yellow. Signal is generated proportionally to the amount of bound analyte and the intensity is measured at 450 nm. Optionally, instead of the endpoint reading, development of TMB can be recorded kinetically at 600 nm.


    As of October 2019, this kit was reformulated with new antibodies to maintain continued long term supply.

  • 说明

    Estimated sensitivity: Phospho-JNK1/2 (Thr183/Tyr185): 0.5 ng/mL (tested with recombinant protein), Total JNK1/2: 0.1 ng/mL (tested with recombinant protein)
    Range: Phospho-JNK1/2 (Thr183/Tyr185): 1-100 ng/mL, Total JNK1/2: 0.2-20 ng/mL

    Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
    It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.

  • 经测试应用

    适用于: ELISAmore details

  • 平台

    Microplate

性能

应用

The Abpromise guarantee

Abpromise™承诺保证使用ab176662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
ELISA
Use at an assay dependent concentration.
说明
ELISA
Use at an assay dependent concentration.

图片

  • Typical cell lysate dilution series
    Typical cell lysate dilution series

    Example of a typical JNK1/2 (pT183/Y185) and JNK1/2 (Total) cell lysate dilution series. Background-subtracted data values (mean +/- SD) are graphed.

  • Typical recombinant protein standard curve
    Typical recombinant protein standard curve

    Example of a typical JNK1/2 (pT183/Y185) recombinant protein standard curve. The proportion of total protein that is phosphorylated is unknown - data is indicative only. Background-subtracted data values (mean +/- SD) are graphed.

  • Linearity of dilution
    Linearity of dilution

    Linearity of dilution in representative sample matrices. Cellular lysates were prepared at 3 concentrations in common media containing 1X Cell Extraction Buffer PTR. Data from duplicate measurements of JNK1/2 (pT183/Y185) are normalized and plotted.

  • Comparison of JNK1/2 expression in different cell lines
    Comparison of JNK1/2 expression in different cell lines

    Cell line analysis for Total JNK1/2 from 200 µg/mL preparations of cell extracts. Data from triplicate measurements (mean +/- SD) are plotted and compared to 1X Cell Extraction Buffer PTR (zero).

  • JNK1/2 (pT183/Y185) phosphorylation in response to anisomycin treatment
    JNK1/2 (pT183/Y185) phosphorylation in response to anisomycin treatment

    Induction of JNK1/2 (pT183/Y185) phosphorylation in HeLa cells in response to anisomycin treatment. HeLa cells were cultured in 96-well tissue culture plates and treated (30 min) with a dose-range of anisomycin before cell lysis. Data from quadruplicate measurements of JNK1/2 (pT183/Y185) are plotted and compared against Total JNK1/2 protein levels. Comparative JNK1/2 (pT183/Y185) and JNK1/2 (Total) data also shown by Western Blot.

  • Sandwich ELISA - JNK 1/2 (pT183/Y185 + Total) ELISA Kit (ab176662)
    Sandwich ELISA - JNK 1/2 (pT183/Y185 + Total) ELISA Kit (ab176662)
    SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
  • Sandwich ELISA - JNK 1/2 (pT183/Y185 + Total) ELISA Kit (ab176662)
    Sandwich ELISA - JNK 1/2 (pT183/Y185 + Total) ELISA Kit (ab176662)
    To learn more about the advantages of SimpleStep ELISA® kits see here.

文献 (12)

发表研究结果有使用 ab176662？请让我们知道，以便我们可以引用本数据表中的参考文章。

ab176662 被引用在 12 文献中.

  • Yin Y  et al. The topoisomerase inhibitor CPT-11 prevents the growth and metastasis of lung cancer cells in nude mice by inhibiting EGFR/MAPK signaling pathway. Heliyon 9:e15805 (2023). PubMed: 37251857
  • Yang Q  et al. Aged black garlic extract inhibits the growth of estrogen receptor-positive breast cancer cells by downregulating MCL-1 expression through the ROS-JNK pathway. PLoS One 18:e0286454 (2023). PubMed: 37352173
  • Sun J  et al. Cayratia japonica Prevents Ulcerative Colitis by Promoting M2 Macrophage Polarization through Blocking the TLR4/MAPK/NF-κB Pathway. Mediators Inflamm 2022:1108569 (2022). PubMed: 36619207
  • Mahnashi MH  et al. Design, Synthesis, and Biological Evaluation of a Novel VEGFR-2 Inhibitor Based on a 1,2,5-Oxadiazole-2-Oxide Scaffold with MAPK Signaling Pathway Inhibition. Pharmaceuticals (Basel) 15:N/A (2022). PubMed: 35215358
  • Wang Z  et al. LncRNA LINC01134 Contributes to Radioresistance in Hepatocellular Carcinoma by Regulating DNA Damage Response via MAPK Signaling Pathway. Front Pharmacol 12:791889 (2021). PubMed: 35173610
View all Publications for this product

客户评价及客户问答

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1-2 of 2 Abreviews or Q&A

Test of ab176662 on pancreatic tissue that have undergo cold ischemia

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
We are actually working on the preservation of rat pancreas from ischemia-reperfusion injury that occurs during the retrieval of the organ before a transplantation to a recipient. We are looking for strong ischemia markers and we know from literature that JNK activation contributes to myocardial IR injury after prolonged hypothermic storage (Vassalli G et al., 2012. Journal of Transplantation).
We extracted cytoplasmic proteins from frozen pancreas that have undergo different time of cold ischemia : 0, 2, 4, 6, 8, 10, 12 and 18h. Proteins were extracted using an homemade lysis buffer (20mM Tris, 1mM EDTA, 1mM EGTA, 150 mM NaCl, 0.5% Triton-X-100 and a protease inhibitor cocktail (Complete™ ULTRA Tablets, EDTA-free, glass vials Protease Inhibitor Cocktail, Roche) and were frozen at -80°C. A BCA test was performed to determine proteins concentration in our sample and 70µg of pancreatic proteins were dilute in 50µl of 1X cell extraction buffer PTR for the assay. OD were read with Glomax from Promega.
OD 450nm P-JNK1/2 JNK1/2 total P-JNK1/2 - blk JNK1/2 total - blk P-JNK/JNK
0h ischemia n1 0,219 0,477 0,151 0,426 0,355
4h ischemia n1 0,330 4,445 0,262 4,393 0,060
6h ischemia n1 0,361 4,388 0,294 4,337 0,068
8h ischemia n1 1,311 4,519 1,244 4,468 0,278
10h ischemia n1 0,476 0,555 0,409 0,504 0,811
12h ischemia n1 0,554 4,446 0,486 4,395 0,111
18h ischemia n1 0,431 0,305 0,363 0,254 1,431
0h ischemia n2 0,398 0,552 0,330 0,501 0,659
4h ischemia n2 0,382 4,434 0,315 4,383 0,072
6h ischemia n2 0,432 4,241 0,365 4,190 0,087
8h ischemia n2 0,339 0,529 0,272 0,478 0,568
10h ischemia n2 1,891 4,644 1,823 4,592 0,397
12h ischemia n2 1,034 0,342 0,966 0,291 3,319
18h ischemia n2 0,337 0,199 0,269 0,148 1,823
0h ischemia n3 0,370 4,117 0,302 4,065 0,074
4h ischemia n3 0,289 4,449 0,222 4,397 0,050
6h ischemia n3 0,309 4,545 0,241 4,494 0,054
8h ischemia n3 0,641 4,468 0,573 4,416 0,130
10h ischemia n3 0,412 1,657 0,344 1,606 0,214
12h ischemia n3 0,262 4,376 0,195 4,325 0,045
18h ischemia n3 0,257 4,425 0,189 4,374 0,043
Blank 0,068 0,051
mean P-JNK/JNK SEM
0H ischemia 0,363 0,169
4H ischemia 0,061 0,006
6H ischemia 0,069 0,010
8H ischemia 0,325 0,129
10H ischemia 0,474 0,176
12H ischemia 1,158 1,080
18H ischemia 1,099 0,540
We obtained concording results with literature : an increase of JNK activation after 8h of cold ischemia and beyond. The kit is working on rat pancreatic tissue.
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. Florent Klack

Verified customer

提交于 Nov 24 2017

The inhibition of JNK in rat bone marrow stem cells by Ruthenium Red.

 Excellent Excellent 5/5 (Ease of Use)
Abreviews
abreview image
Abtrial CODE: ABTRIAL-EV7WD
Methods
1. Wash the 6 well plate with PBS after ultrasound exposure.
2. Lyse the cells with 0.5mL of freshly prepared Lysis buffer per well, with shaking (~300 rpm) at room temp for 10 minutes.
3. Aliquot supernatant (this is the soluble cell extract) to clean, chilled tubes on ice and store samples at -80°C. Minimize freeze/thaw cycles.
4. Determine desired number of microplate strips. Remove unused strips from frame and return to storage pouch and seal.
5. Add 50 μL/well of lysate to the microplate.
6. Add 50 μL/well Lysis Mix (negative control) and Control Lysates (positive control) to separate wells for assay controls if desired.
7. Add 50 μL/well of Antibody Mix to the wells. Cover the microplate with adhesive seal and incubate for 1 hr at room temp on a microplate shaker (400 rpm).
8. Wash wells with 350 μL/well 1X Wash Buffer (repeat 3 times). After final wash, remove any remaining wash solution from wells.
9. Add μL/well TMB and incubate in the dark on a plate shaker set to 400rpm.
10. Add 100 µL/well Stop Solution, and mix briefly (1 min) on a microplate shaker.
11. Record the OD at 450 nm.

Results
Phosphorylation of JNK was not significantly affected by Ruthenium Red after ultrasound exposure in rat BMSCs.
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.

MS. XXXXXX XXXXXX

Verified customer

提交于 Jul 15 2015

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