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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Tested Applications & Species
See all applications and species dataApplication Species ICC/IF MouseRatHumanWB Human
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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.
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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.
One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.
Learn more here.
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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Related Products
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- Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)
应用
The Abpromise guarantee
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.
Tested applications are guaranteed to work and covered by our Abpromise guarantee.
Predicted to work for this combination of applications and species but not guaranteed.
Does not work for this combination of applications and species.
| 应用 | Species |
|---|---|
| ICC/IF |
Mouse
Rat
Human
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| WB |
Human
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| All applications |
Horse
Chicken
Guinea pig
Hamster
Cat
Dog
Pig
Xenopus laevis
Fish
Monkey
Zebrafish
Baboon
Xenopus tropicalis
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| 应用 | Ab评论 | 说明 |
|---|---|---|
| WB | (91) |
1/500 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 40.2 kDa).
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| ICC/IF | (1) |
Use a concentration of 1 - 5 µg/ml.
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| 说明 |
|---|
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WB
1/500 - 1/10000. Detects a band of approximately 36 kDa (predicted molecular weight: 40.2 kDa). |
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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
数据表及文件
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SDS download
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Datasheet download
文献 (2434)
ab8245 被引用在 2434 文献中.
- 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
- Zhao W et al. Splicing factor derived circular RNA circUHRF1 accelerates oral squamous cell carcinoma tumorigenesis via feedback loop. Cell Death Differ 27:919-933 (2020). PubMed: 31570856
- Lorenz DA et al. Direct RNA sequencing enables m6A detection in endogenous transcript isoforms at base-specific resolution. RNA 26:19-28 (2020). PubMed: 31624092
- Zhang Q et al. Apoptotic SKOV3 cells stimulate M0 macrophages to differentiate into M2 macrophages and promote the proliferation and migration of ovarian cancer cells by activating the ERK signaling pathway. Int J Mol Med 45:10-22 (2020). PubMed: 31746376
- He ZH et al. The nuclear transcription factor FoxG1 affects the sensitivity of mimetic aging hair cells to inflammation by regulating autophagy pathways. Redox Biol 28:101364 (2020). PubMed: 31731101