重组Anti-IRS2抗体[EPR904(2)] (ab134101)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR904(2)] to IRS2
- Suitable for: Flow Cyt (Intra), WB, IHC-P, ICC/IF
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
概述
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产品名称
Anti-IRS2抗体[EPR904(2)]
参阅全部 IRS2 一抗 -
描述
兔单克隆抗体[EPR904(2)] to IRS2 -
宿主
Rabbit -
经测试应用
适用于: Flow Cyt (Intra), WB, IHC-P, ICC/IFmore details -
种属反应性
与反应: Mouse, Rat, Human -
免疫原
Synthetic peptide within Human IRS2 aa 1300 to the C-terminus (C terminal). The exact sequence is proprietary.
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阳性对照
- WB: HEK293, HEK293 (treated with insulin), and NIH/3T3 (treated with insulin) and SH-SY5Y cell lysates; A375 (human malignant melanoma epithelial cell) and A549 whole cell lysates IHC-P: Human kidney, breast and muscle tissues and rat kidney tissue. ICC/IF: SH-SY5Y cells. HEK293 WT cells (HEK293-IRS2 KO used as a negative cell line). Flow Cyt (intra): HeLa cells.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
性能
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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. Stable for 12 months at -20°C. -
存储溶液
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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纯度
Protein A purified -
克隆
单克隆 -
克隆编号
EPR904(2) -
同种型
IgG -
研究领域
相关产品
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Recombinant Protein
应用
The Abpromise guarantee
Abpromise™承诺保证使用ab134101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 |
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Flow Cyt (Intra) |
1/100 - 1/1000.
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB |
1/1000 - 1/10000. Detects a band of approximately 170-185 kDa (predicted molecular weight: 137 kDa).
Although some papers support the expression in liver (PMID: 30202052), A549 (PMID: 30988063), NCI-H1299 (PMID: 30988063), LADMAC (PMID: 29115630), MDA-MB-231 (PMID: 29685905) and MEF (PMID: 30679431), ab134101 can’t detect the target band in these samples, even at the dilution of 1:200 |
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IHC-P |
1/50 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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ICC/IF |
1/300.
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说明 |
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Flow Cyt (Intra)
1/100 - 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/1000 - 1/10000. Detects a band of approximately 170-185 kDa (predicted molecular weight: 137 kDa). Although some papers support the expression in liver (PMID: 30202052), A549 (PMID: 30988063), NCI-H1299 (PMID: 30988063), LADMAC (PMID: 29115630), MDA-MB-231 (PMID: 29685905) and MEF (PMID: 30679431), ab134101 can’t detect the target band in these samples, even at the dilution of 1:200 |
IHC-P
1/50 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
ICC/IF
1/300. |
靶标
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功能
May mediate the control of various cellular processes by insulin. -
序列相似性
Contains 1 IRS-type PTB domain.
Contains 1 PH domain. -
翻译后修饰
Phosphorylated upon DNA damage, probably by ATM or ATR. -
细胞定位
Cytoplasm > cytosol. - Information by UniProt
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数据库链接
- Entrez Gene: 8660 Human
- Entrez Gene: 384783 Mouse
- Entrez Gene: 29376 Rat
- Omim: 600797 Human
- SwissProt: Q9Y4H2 Human
- SwissProt: P81122 Mouse
- Unigene: 442344 Human
- Unigene: 407207 Mouse
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别名
- Insulin receptor substrate 2 antibody
- IRS 2 antibody
- IRS-2 antibody
see all
图片
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All lanes : Anti-IRS2 antibody [EPR904(2)] (ab134101) at 1/1000 dilution
Lane 1 : Wild-type HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 2 : IRS2 knockout HEK-293 (Human epithelial cell line from embryonic kidney) whole cell lysate
Lane 3 : A-375 cell lysate
Lane 4 : A549 (Human lung carcinoma cell line) whole cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 137 kDa
Observed band size: 160 kDa why is the actual band size different from the predicted?Lanes 1 - 4: Merged signal (red and green). Green - ab134101 observed at 160 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.
ab134101 was shown to react with IRS2 in wild-type HEK-293 cells in western blot with loss of signal observed in IRS2 knockout sample. Wild-type and IRS2 knockout HEK-293 cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% milk in TBS-T (0.1% Tween®) before incubation with ab134101 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
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ab134101 staining IRS2 in wild-type HEK293 cells (top panel) and IRS2 knockout HEK293 cells (bottom panel). 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 with ab134101 at 1/500 and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at +4°C, followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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Immunocytochemistry/Immunofluorescence analysis of SH-SY5Y cells labelling IRS2 with purified ab134101 at 1/300. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.
Control 1: primary antibody (1/300) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000).
Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/1000).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS2 antibody [EPR904(2)] (ab134101)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat kidney tissue labelling IRS2 with purified ab134101 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS2 antibody [EPR904(2)] (ab134101)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney tissue labelling IRS2 with purified ab134101 at 1/500. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS2 antibody [EPR904(2)] (ab134101)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human muscle tissue labelling IRS2 with unpurified ab134101 at a dilution of 1/50.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IRS2 antibody [EPR904(2)] (ab134101)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast tissue labelling IRS2 with unpurified ab134101 at a dilution of 1/50.
Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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All lanes : Anti-IRS2 antibody [EPR904(2)] (ab134101) at 1/1000 dilution
Lane 1 : Rat liver lysates
Lane 2 : Human liver lysates
Lane 3 : A375 (human malignant melanoma epithelial cell) whole cell lysates
Lane 4 : A549 (human lung carcinoma epithelial cell) whole cell lysates
Lane 5 : NCI-H1299 (human lung carcinoma epithelial cell) whole cell lysates
Lane 6 : LADMAC (Mouse bone marrow monocyte macrophage) whole cell lysates
Lane 7 : MDA-MB-231 (human breast adenocarcinoma epithelial cell) whole cell lysates
Lane 8 : MEF (Mouse embryonic fibroblast) whole cell lysates
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution (Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated)
Developed using the ECL technique.
Predicted band size: 137 kDa
Observed band size: 170-185 kDa why is the actual band size different from the predicted?Blocking/Diluting buffer and concentration: 5% NFDM/TBST.
Exposure time:
Left image: 180 seconds
Right image: 40 seconds
Although some papers support the expression in liver (PMID: 30202052), A549 (PMID: 30988063), NCI-H1299 (PMID: 30988063), LADMAC (PMID: 29115630), MDA-MB-231 (PMID: 29685905) and MEF (PMID: 30679431), ab134101 can’t detect the target band in these samples, even at the dilution of 1:200.
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Anti-IRS2 antibody [EPR904(2)] (ab134101) at 1/5000 dilution (purified) + NIH/3T3 whole cell lysate - treated with insulin at 20 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 137 kDa
Observed band size: 170-185 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
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All lanes : Anti-IRS2 antibody [EPR904(2)] (ab134101) at 20 µg (purified)
Lane 1 : HEK293 whole cell lysate - untreated
Lane 2 : HEK293 whole cell lysate - treated with insulin
Lane 3 : SH-SY5Y whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution
Predicted band size: 137 kDa
Observed band size: 170-185 kDa why is the actual band size different from the predicted?Blocking and dilution buffer: 5% NFDM/TBST.
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Intracellular Flow Cytometry analysis of HeLa cells labelling IRS2 with purified ab134101 at 1/120 (red). Cells were fixed with 2% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.
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Overlay histogram showing HeLa cells stained with unpurified ab134101 (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 (unpurified ab134101, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa 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
数据表及文件
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SDS download
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Datasheet download
文献 (30)
ab134101 被引用在 30 文献中.
- Selven H et al. High Expression of IRS-1, RUNX3 and SMAD4 Are Positive Prognostic Factors in Stage I-III Colon Cancer. Cancers (Basel) 15:N/A (2023). PubMed: 36900240
- Chen Q et al. Steatotic hepatocyte-derived extracellular vesicles promote β-cell apoptosis and diabetes via microRNA-126a-3p. Liver Int 43:2560-2570 (2023). PubMed: 37337778
- Ren J et al. Amelioration of gamma irradiation-induced salivary gland damage in mice using melatonin. J Pineal Res 75:e12897 (2023). PubMed: 37391878
- Omar IS et al. MPA alters metabolic phenotype of endometrial cancer-associated fibroblasts from obese women via IRS2 signaling. PLoS One 17:e0270830 (2022). PubMed: 35816477
- Liu X et al. β-Carotene regulates glucose transport and insulin resistance in gestational diabetes mellitus by increasing the expression of SHBG. Clin Exp Pharmacol Physiol 49:1307-1318 (2022). PubMed: 35986631