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Anti-GAPDH抗体[6C5] - Loading Control (ab8245)

  • Datasheet
  • SDS
Reviews (95)Q&A (10)References (2434)

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Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)
  • Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)
  • Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)
  • Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)
  • Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)
  • Immunocytochemistry/ Immunofluorescence - 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

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概述

  • 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.

  • 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.

  • 阳性对照

    • ICC/IF: HeLa cells, NIH3T3 cells, SV40LT-SMC cells. WB: HeLa, A431, Jurkat, HEK-293, Raji whole cell lysate.
  • 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

  • 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

    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Huntington's disease
    • Neuroscience
    • Neurology process
    • Neurodegenerative disease
    • Alzheimer's disease
    • Other
    • Isotype/Loading Controls
    • Loading Controls
    • GAPDH
    • Signal Transduction
    • Metabolism
    • Energy Metabolism
    • Cancer
    • Cancer Metabolism
    • Metabolic signaling pathway
    • Metabolism of carbohydrates
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Carbohydrate metabolism
    • Metabolism
    • Pathways and Processes
    • Metabolic signaling pathways
    • Energy transfer pathways
    • Energy Metabolism
    • Metabolism
    • Types of disease
    • Neurodegenerative disease
    • Metabolism
    • Types of disease
    • Cancer

相关产品

  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (HRP) (ab205719)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Mouse IgG1, kappa monoclonal [15-6E10A7] - Isotype Control (ab170190)
  • Positive Controls

    • HeLa nuclear extract lysate (ab14655)
    • HeLa whole cell lysate (ab150035)
    • HeLa whole cell lysate (ab29545)
    • Jurkat whole cell lysate (ab30128)
    • Jurkat whole cell lysate (ab7899)
    • HEK293 whole cell lysate (ab7902)
    • A431 whole cell lysate (ab7909)
  • Recombinant Protein

    • Recombinant Mouse GAPDH protein (ab202148)
    • Recombinant Human GAPDH protein (ab77109)
    • Recombinant human GAPDH protein (Active) (ab82633)
  • Related Products

    • Prestained Protein Ladder - Broad molecular weight (10-245 kDa) (ab116028)
    • Anti-SDHB antibody [21A11AE7] (ab14714)
    • Anti-SDHA antibody [2E3GC12FB2AE2] (ab14715)
    • Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291)
    • Anti-beta Actin antibody [mAbcam 8224] - Loading Control (ab8224)
    • Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484)

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

  • 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.
  • Target information above from: UniProt accession P04406 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: 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
    • Entrez Gene: 685186 Rat
    • Entrez Gene: 380259 Xenopus laevis
    • Entrez Gene: 317743 Zebrafish
    • Omim: 138400 Human
    • SwissProt: Q9N2D5 Cat
    • SwissProt: P00356 Chicken
    • SwissProt: Q28259 Dog
    • SwissProt: P04406 Human
    • SwissProt: P16858 Mouse
    • SwissProt: P00355 Pig
    • SwissProt: P04797 Rat
    • SwissProt: P51469 Xenopus laevis
    • SwissProt: Q5XJ10 Zebrafish
    • Unigene: 544577 Human
    • Unigene: 592355 Human
    • Unigene: 598320 Human
    • Unigene: 304088 Mouse
    • Unigene: 309092 Mouse
    • Unigene: 317779 Mouse
    • Unigene: 343110 Mouse
    • Unigene: 392463 Mouse
    • Unigene: 392480 Mouse
    • Unigene: 414470 Mouse
    • Unigene: 458138 Mouse
    • Unigene: 458416 Mouse
    • Unigene: 475698 Mouse
    • Unigene: 129558 Rat
    • Unigene: 91450 Rat
    • Unigene: 995 Xenopus laevis
    • Unigene: 35640 Zebrafish
    see all
  • 别名

    • 38 kDa BFA-dependent ADP-ribosylation substrate antibody
    • aging associated gene 9 protein antibody
    • Aging-associated gene 9 protein antibody
    • BARS-38 antibody
    • cb609 antibody
    • EC 1.2.1.12 antibody
    • Epididymis secretory sperm binding protein Li 162eP antibody
    • G3P_HUMAN antibody
    • G3PD antibody
    • G3PDH antibody
    • GAPD antibody
    • GAPDH antibody
    • Glyceraldehyde 3 phosphate dehydrogenase antibody
    • glyceraldehyde 3-PDH antibody
    • Glyceraldehyde-3-phosphate dehydrogenase antibody
    • HEL-S-162eP antibody
    • KNC-NDS6 antibody
    • MGC102544 antibody
    • MGC102546 antibody
    • MGC103190 antibody
    • MGC103191 antibody
    • MGC105239 antibody
    • MGC127711 antibody
    • MGC88685 antibody
    • OCAS, p38 component antibody
    • OCT1 coactivator in S phase, 38-KD component antibody
    • peptidyl cysteine S nitrosylase GAPDH antibody
    • Peptidyl-cysteine S-nitrosylase GAPDH antibody
    • wu:fb33a10 antibody
    see all

图片

  • Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)
    Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)This image is courtesy of an anonymous Abreview
    All 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

    See Abreview

  • Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)
    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).

  • Western blot - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)
    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)
    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)
    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).

  • Immunocytochemistry/ Immunofluorescence - Anti-GAPDH antibody [6C5] - Loading Control (ab8245)
    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

  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

数据表及文件

    • Datasheet
    • SDS
  • 文献 (2434)

    发表研究结果有使用 ab8245?请让我们知道,以便我们可以引用本数据表中的参考文章。

    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
    View all Publications for this product

    客户评价及客户问答

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    Western blot abreview for Anti-GAPDH antibody [6C5] - Loading Control

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    Abreviews
    Abreviews
    abreview image
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (hek293)
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Loading amount
    25 µg
    Specification
    hek293
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Read More

    Dr. Robert Kupp

    Verified customer

    提交于 Oct 19 2020

    Western blot abreview for Anti-GAPDH antibody [6C5] - Loading Control

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    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (U87 cells)
    Gel Running Conditions
    Non-reduced Non-Denaturing (Native) (4-20% gel)
    Loading amount
    15 µg
    Treatment
    bevacizumab 2mg/ml for 24hrs, 48hrs
    Specification
    U87 cells
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    提交于 May 11 2020

    Western blot abreview for Anti-GAPDH antibody [6C5] - Loading Control

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    Application
    Western blot
    Sample
    Rat Cell lysate - whole cell (Hippocampal culture)
    Gel Running Conditions
    Reduced Denaturing (4-15% gradient)
    Loading amount
    20 µg
    Specification
    Hippocampal culture
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    提交于 Jan 31 2020

    Western blot abreview for Anti-GAPDH antibody [6C5] - Loading Control

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    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (Cerebellum)
    Gel Running Conditions
    Reduced Denaturing (4-15% gradient)
    Loading amount
    20 µg
    Specification
    Cerebellum
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Read More

    Abcam user community

    Verified customer

    提交于 Jan 31 2020

    Western blot abreview for Anti-GAPDH antibody [6C5] - Loading Control

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    abreview image
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    Sample
    Human Cell lysate - whole cell (HEK293)
    Gel Running Conditions
    Reduced Denaturing (4-15% gradient)
    Loading amount
    10 µg
    Specification
    HEK293
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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    Sample
    Rat Tissue lysate - whole (Brain tissue)
    Gel Running Conditions
    Reduced Denaturing (8% SDS Running gel)
    Loading amount
    80 µg
    Treatment
    10mg ketamine
    Specification
    Brain tissue
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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    提交于 Nov 12 2019

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    Human Tissue lysate - whole (skin tissue)
    Gel Running Conditions
    Reduced Denaturing (10% gel)
    Loading amount
    30 µg
    Treatment
    UV B (100mJ/cm2)
    Specification
    skin tissue
    Blocking step
    BSA as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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    Sample
    Mouse Cell lysate - other (MEF (cytosol fraction))
    Gel Running Conditions
    Reduced Denaturing (12)
    Loading amount
    10 µg
    Specification
    MEF (cytosol fraction)
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
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    Sample
    Human Cell lysate - whole cell (H1299 cells)
    Gel Running Conditions
    Reduced Denaturing (12)
    Loading amount
    25 µg
    Specification
    H1299 cells
    Blocking step
    Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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    Sample
    Mouse Tissue lysate - whole (Brain, heart, liver)
    Gel Running Conditions
    Reduced Denaturing (12% gel)
    Loading amount
    25 µg
    Specification
    Brain, heart, liver
    Blocking step
    Licor Odyssey Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 25°C
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