• 概述

• 产品描述 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. Modulates the organization and assembly of the cytoskeleton. Facilitates the CHP1-dependent microtubule and membrane associations through its ability to stimulate the binding of CHP1 to microtubules. 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. Component of the GAIT (gamma interferon-activated inhibitor of translation) complex which mediates interferon-gamma-induced transcript-selective translation inhibition in inflammation processes. Upon interferon-gamma treatment assembles into the GAIT complex which binds to stem loop-containing GAIT elements in the 3'-UTR of diverse inflammatory mRNAs and suppresses their translation.
• 产品名称 Anti-GAPDH Mouse Monoclonal Loading Control Antibody [12D6]
• 分子量 Predicted band size: 36 kDa
• 种属反应性 Human,Mouse,Rat,Zebrafish,Escherichia coli
• 验证应用 WB,IF-Cell,IHC-P
• 抗体类型 小鼠单抗
• 免疫原 Synthetic peptide within human GAPDH aa 180-220.
• 偶联 Non-conjugated

• 性能

• 形态 Liquid
• 浓度 2 mg/mL.
• 存放说明 Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
• 存储缓冲液 1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
• 亚型 IgG1
• 纯化方式 Protein G affinity purified.
• 亚细胞定位 Cytoplasm, Nucleus, Membrane.
• 数据链接 SwissProt: P04406 Human
  SwissProt: P16858 Mouse
  SwissProt: P04797 Rat
  
• 其它名称
  • 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-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
  more

• 应用

  WB: 1:2000-1:5,000
  IF-Cell: 1:200
  IHC-P: 1:600

• 

  Fig1: Western blot analysis of GAPDH on different cells lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (M1310-2, 1/1000) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.
  Positive control:
  Lane 1: Hela cell lysate
  Lane 2: A549 cell lysate
  Lane 3: HepG2 cell lysate
  Lane 4: PC-12 cell lysate
  Lane 5: F9 cell lysate

  

  Fig2: Western blot analysis of GAPDH on zebrafish tissue lysates. Proteins were transferred to a PVDF membrane and blocked with 5% BSA in PBS for 1 hour at room temperature. The primary antibody (M1310-2, 1/500) was used in 5% BSA at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:5,000 dilution was used for 1 hour at room temperature.

  

  Fig3: Western blot analysis of GAPDH on different lysates with Mouse anti-GAPDH antibody (M1310-2) at 1/500 dilution.
  
  Lane 1: Escherichia coli lysate
  Lane 2: Escherichia coli lysate
  
  Lysates/proteins at 10 µg/Lane.
  
  Predicted band size: 36 kDa
  Observed band size: 36 kDa
  
  Exposure time: 1 minute;
  
  4-20% SDS-PAGE gel.
  
  Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (M1310-2) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:150,000 dilution was used for 1 hour at room temperature.

  

  Fig4: ICC staining of GAPDH in D3 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (M1310-2, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor®555 Goat anti-Mouse IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).

  

  Fig5: ICC staining of GAPDH in A431 cells (red). Formalin fixed cells were permeabilized with 0.1% Triton X-100 in TBS for 10 minutes at room temperature and blocked with 1% Blocker BSA for 15 minutes at room temperature. Cells were probed with the primary antibody (M1310-2, 1/100) for 1 hour at room temperature, washed with PBS. Alexa Fluor®555 Goat anti-Mouse IgG was used as the secondary antibody at 1/100 dilution. The nuclear counter stain is DAPI (blue).

  

  Fig6: Immunohistochemical analysis of paraffin-embedded human thyroid carcinoma tissue with Mouse anti-GAPDH antibody (M1310-2) at 1/600 dilution.
  
  The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1310-2) at 1/600 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

  

  Fig7: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Mouse anti-GAPDH antibody (M1310-2) at 1/600 dilution.
  
  The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1310-2) at 1/600 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

• 背景文献

• 1. Cheng, Y., Hou, T., Ping, J., Chen, T., & Yin, B. (2018). LMO3 promotes hepatocellular carcinoma invasion, metastasis and anoikis inhibition by directly interacting with LATS1 and suppressing Hippo signaling. J Exp Clin Cancer Res, 37(1), 228.
  
  2. Liu, J., Wang, Y., Song, L., Zeng, L., Yi, W., Liu, T., . . . Cong, Y. S. (2017). A critical role of DDRGK1 in endoplasmic reticulum homoeostasis via regulation of IRE1alpha stability. Nat Commun, 8, 14186.
  
  3. Jiang, Y., Tian, M., Lin, W., Wang, X., & Wang, X. (2018). Protein Kinase Serine/Threonine Kinase 24 Positively Regulates Interleukin 17-Induced Inflammation by Promoting IKK Complex Activation. Front Immunol, 9, 921.
  
  4. Bin, G., Jiarong, Z., Shihao, W., Xiuli, S., Cheng, X., Liangbiao, C., & Ming, Z. (2012). Aire promotes the self-renewal of embryonic stem cells through Lin28. Stem Cells Dev, 21(15), 2878-2890.
  
  5. Liu, J., Wang, Y., Song, L., et al. Cong, Y. S. (2017). A critical role of DDRGK1 in endoplasmic reticulum homoeostasis via regulation of IRE1alpha stability. Nat Commun, 8, 14186. doi: 10.1038/ncomms14186
  
  6. Zhang, D., Zhao, Q., Sun, H., Yin, L., Wu, J., Xu, J., . . . Liang, C. (2016). Defective autophagy leads to the suppression of stem-like features of CD271(+) osteosarcoma cells. J Biomed Sci, 23(1), 82. doi: 10.1186/s12929-016-0297-5

• Publishing research using Anti-GAPDH Mouse Monoclonal Loading Control Antibody? Please let us know so that we can cite the reference in this datasheet.

• PMID:
  35036955 Optimized protocol to detect protein UFMylation in cells and in vitro via immunoblotting Author: Zhou, J., Liang, Q., Dong, M., Ma, X., Jin, Y., Guan, D., Liu, J., Wang, M., & Cong, Y. S. Published: 2022 Application: WB Species: Human Journal: STAR Protocols
• PMID:
  36016966 Inhibition of pyruvate dehydrogenase kinase improves carbohydrate utilization in Nile tilapia by regulating PDK2/4-PDHE1α axis and insulin sensitivity Author: Luo, Y., Zhou, W., Li, R., Limbu, S. M., Qiao, F., Chen, L., Zhang, M., & Du, Z. Y. Published: 2022 Application: WB Species: Nile tilapia Journal: Anim Nutr. IF: 5.285
• PMID:
  36033499 Salvia chinensis Benth Inhibits Triple-Negative Breast Cancer Progression by Inducing the DNA Damage Pathway Author: Wang, K. N., Hu, Y., Han, L. L., Zhao, S. S., Song, C., Sun, S. W., Lv, H. Y., Jiang, N. N., Xv, L. Z., Zhao, Z. W., & Li, M. Published: 2022 Application: WB Species: Mouse Journal: Front Oncol. IF: 5.738
• PMID:
  36120581 Ufl1 deficiency causes skin pigmentation by up-regulation of Endothelin-1 Author: Wang, K., Xu, H. N., Wang, Y. W., Mao, J., Liu, D., Zhu, X. J., Cong, Y. S., & Wang, M. Published: 2022 Application: WB Species: Mouse Journal: Front Cell Dev Biol. IF: 6.081
• PMID:
  34148841 Ufl1 deficiency causes kidney atrophy associated with disruption of endoplasmic reticulum homeostasis Author: Zhou, Y., Ye, X., Zhang, C., Wang, J., Guan, Z., Yan, J., Xu, L., Wang, K., Guan, D., Liang, Q., Mao, J., Zhou, J., Zhang, Q., Wu, X., Wang, M., Cong, Y. S., & Liu, J. Published: 2021 Application: WB Species: Human Journal: J Genet Genomics IF: 4.275
• PMID:
  32468020 Upregulation of miRNA-301a-3p promotes tumor progression in gastric cancer by suppressing NKRF and activating NF-κB signaling Author: Xujun He Published: 2020 Application: WB Species: human Journal: INTERNATIONAL JOURNAL OF ONCOLOGY IF: 3.899
• PMID:
  32236608 CD44 inhibition attenuates EGFR signaling and enhances cisplatin sensitivity in human EGFR wild-type non-small-cell lung cancer cells Author: Dandan Yuan;Mingxing Li Published: 2020 Application: WB,IF Species: human Journal: Int J Mol Med. IF: 2.928
• PMID:
  32903879 Integrative Analysis of the IQ Motif-Containing GTPase-Activating Protein Family Indicates That the IQGAP3-PIK3C2B Axis Promotes Invasion in Colon Cancer Author: Yuping Zhu; Xujun He Published: 2020 Application: WB Species: human Journal: OncoTargets and Therapy IF: 3.337
• PMID:
  30989650 Identification of the potential key genes for adipogenesis from human mesenchymal stem cells by RNA‐Seq Author: Xia Yi,Xiaoyuan Xu,and Weidong Li Published: 2019 Application: WB Species: hMSCs Journal: J Cell Physiol IF: 4.522
• PMID:
  28128204 A critical role of DDRGK1 in endoplasmic reticulum homoeostasis via regulation of IRE1a stability Author: Zhenyu Ju & Yu-Sheng Cong Published: 2017 Application: WB Species: HEK293T,MCF7 and HepG2 cells Journal: Nat Commun IF: 11.3

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Specific Protocols

To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.