Anti-GAPDH抗体[6C5] - Loading Control (ab8245)
![Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)](https://www.abcam.cn/ps/products/8/ab8245/Images/ab8245-209020-anti-gapdh-antibody-6c5-loading-control-western-blot.jpg "Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)")
Key features and details
- Mouse monoclonal [6C5] to GAPDH - Loading Control
- Suitable for: WB, ICC/IF
- Reacts with: Mouse, Rat, Human
- Isotype: IgG1
概述
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产品名称
Anti-GAPDH抗体[6C5] - Loading Control
参阅全部 GAPDH 一抗 -
描述
小鼠单克隆抗体[6C5] to GAPDH - Loading Control -
宿主
Mouse -
特异性
This GAPDH antibody can be used as a loading control antibody. GAPDH is a 146 kDa tetramer composed of four 30-40 kDa subunits. There is no cross-reaction with GAPDH from yeast. Preliminary data indicates that the GAPDH antibody- loading control ab8245 recognizes the monomer (36 kDa) and also the dimer forms of GAPDH, but not the tetrameric form of the protein.
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经测试应用
适用于: WB, ICC/IFmore details -
种属反应性
与反应: Mouse, Rat, Human
预测可用于: Horse, Chicken, Guinea pig, Hamster, Cat, Dog, Pig, Xenopus laevis, Fish, Monkey, Zebrafish, Baboon, Xenopus tropicalis
不与反应:
Goat, Cow, Saccharomyces cerevisiae -
免疫原
Rabbit muscle GAPDH.
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阳性对照
- ICC/IF: HeLa cells, NIH3T3 cells, SV40LT-SMC cells. WB: HeLa, A431, Jurkat, HEK-293, Raji whole cell lysate.
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常规说明
This product switched from ascites to tissue culture supernatant on 31 July 2017. Lot numbers higher than [GR291713] will be from tissue culture supernatant.
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
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, as well as customer reviews and Q&As.
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 or -80°C. Avoid freeze / thaw cycle. -
存储溶液
pH: 7.40
Preservative: 0.09% Sodium azide
Constituent: PBS -
Concentration information loading... -
纯度
Protein A purified -
纯化说明
Chromatography on protein A Sepharose -
克隆
单克隆 -
克隆编号
6C5 -
骨髓瘤
Sp2/0 -
同种型
IgG1 -
研究领域
相关产品
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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应用
Our Abpromise guarantee covers the use of ab8245 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB | 1/500 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 40.2 kDa). | |
| ICC/IF | Use a concentration of 1 - 5 µg/ml. |
靶标
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功能
Has both glyceraldehyde-3-phosphate dehydrogenase and nitrosylase activities, thereby playing a role in glycolysis and nuclear functions, respectively. Participates in nuclear events including transcription, RNA transport, DNA replication and apoptosis. Nuclear functions are probably due to the nitrosylase activity that mediates cysteine S-nitrosylation of nuclear target proteins such as SIRT1, HDAC2 and PRKDC (By similarity). Glyceraldehyde-3-phosphate dehydrogenase is a key enzyme in glycolysis that catalyzes the first step of the pathway by converting D-glyceraldehyde 3-phosphate (G3P) into 3-phospho-D-glyceroyl phosphate. -
通路
Carbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5. -
序列相似性
Belongs to the glyceraldehyde-3-phosphate dehydrogenase family. -
翻译后修饰
S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
ISGylated. -
细胞定位
Cytoplasm > cytosol. Nucleus. Cytoplasm > perinuclear region. Membrane. Translocates to the nucleus following S-nitrosylation and interaction with SIAH1, which contains a nuclear localization signal (By similarity). Postnuclear and Perinuclear regions. - Information by UniProt
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数据库链接
- Entrez Gene: 493876 Cat
- Entrez Gene: 374193 Chicken
- Entrez Gene: 403755 Dog
- Entrez Gene: 2597 Human
- Entrez Gene: 100042025 Mouse
- Entrez Gene: 14433 Mouse
- Entrez Gene: 396823 Pig
- Entrez Gene: 24383 Rat
see all -
别名
- 38 kDa BFA-dependent ADP-ribosylation substrate antibody
- aging associated gene 9 protein antibody
- Aging-associated gene 9 protein antibody
see all
图片
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Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)This image is courtesy of an anonymous AbreviewAll lanes : Anti-GAPDH antibody [6C5] - Loading Control (ab8245)
Lane 1 : Mouse hippocampus whole cell lysate
Lane 2 : Rat hippocampus whole cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP-conjugated Rabbit anti-mouse at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 40.2 kDa
Observed band size: 36 kDa why is the actual band size different from the predicted?
Exposure time: 10 seconds
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![Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)](https://www.abcam.cn/ps/products/8/ab8245/Images/ab8245-249031-anti-gapdh-antibody-6c5-loading-control-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)ab8245 staining GAPDH in SV40LT-SMC (Rat SV40-transfected aorta smooth cell line) cells.
The cells were fixed with 4% formaldehyde (10 minutes), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab8245 at 5μg/ml and ab202272 at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) (shown in green). Nuclear DNA was labeled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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![Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)](https://www.abcam.cn/ps/products/8/ab8245/Images/ab8245_1.gif)
Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)Anti-GAPDH antibody [6C5] - Loading Control (ab8245) at 10 µg/ml + Raji (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg
Predicted band size: 40.2 kDa -
![Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)](https://www.abcam.cn/ps/products/8/ab8245/Images/ab8245-14003-anti-gapdh-antibody-6c5-loading-control-western-blot.jpg)
Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)All lanes : Anti-GAPDH antibody [6C5] - Loading Control (ab8245) at 2.5 µg/ml
Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) Nuclear
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : A431 (Human epidermoid carcinoma cell line) cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) cell lysate
Lane 5 : HEK-293 (Human epithelial cell line from embryonic kidney) cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Alexa Fluor anti-mouse at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 40.2 kDa
Observed band size: 37 kDa why is the actual band size different from the predicted?
Fluorescence detection of secondary antibody. -
![Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)](https://www.abcam.cn/ps/products/8/ab8245/Images/ab8245-249030-anti-gapdh-antibody-6c5-loading-control-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)ab8245 staining GAPDH in NIH/3T3 (Mouse embryo fibroblast cell line) cells.
The cells were fixed with 4% formaldehyde (10 minutes) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab8245 at 1 μg/ml and ab202272 at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1 hour with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) (shown in green). Nuclear DNA was labeled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).
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![Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)](https://www.abcam.cn/ps/products/8/ab8245/Images/ab8245-228734-anti-gapdh-antibody-6c5-loading-control-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)ab8245 staining GAPDH in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.
The cells were fixed with 100% methanol (5 minutes) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1 hour. The cells were then incubated with ab8245 at 5 μg/ml and ab6046 at 1 µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1 hour with Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 2 μg/ml (shown in green) and Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed (ab150088) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labeled in blue with DAPI.
Negative controls: 1– Rabbit primary antibody and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.
实验方案
文献 (2434)
ab8245 被引用在 2434 文献中.
- He B et al. MicroRNA-326 decreases tau phosphorylation and neuron apoptosis through inhibition of the JNK signaling pathway by targeting VAV1 in Alzheimer's disease. J Cell Physiol 235:480-493 (2020). PubMed: 31385301
- Sun H et al. miR-4429 sensitized cervical cancer cells to irradiation by targeting RAD51. J Cell Physiol 235:185-193 (2020). PubMed: 31190335
- Li L et al. XIST/miR-376c-5p/OPN axis modulates the influence of proinflammatory M1 macrophages on osteoarthritis chondrocyte apoptosis. J Cell Physiol 235:281-293 (2020). PubMed: 31215024
- Xu J & Zhang J LncRNA TP73-AS1 is a novel regulator in cervical cancer via miR-329-3p/ARF1 axis. J Cell Biochem 121:344-352 (2020). PubMed: 31232491
- Doccini S et al. Proteomic and functional analyses in disease models reveal CLN5 protein involvement in mitochondrial dysfunction. Cell Death Discov 6:18 (2020). WB ; Human . PubMed: 32257390
