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產品編號	bsm-33033M
英文名稱	Mouse　Anti-GAPDH-Loading Control　antibody
中文名稱	3-磷酸甘油醛脫氫酶（內參）單克隆抗體
別　　　　名	38 kDa BFA-dependent ADP-ribosylation substrate; Aging-associated gene 9 protein; BARS-38; cb609; EC 1.2.1.12; G3PD; G3PDH; GAPD; Glyceraldehyde 3 phosphate dehydrogenase;Glyceraldehyde 3 phosphate dehydrogenase liver;Glyceraldehyde 3 phosphate dehydrogenase muscle; KNC-NDS6; MGC102544; MGC102546; MGC103190; MGC103191; MGC105239; MGC127711; MGC88685; OCAS, p38 component; OCT1 coactivator in S phase, 38-KD component; wu:fb33a10.
	Specific References　　(55)　　　　　\|　　　　　bsm-33033M has been referenced in 55 publications. [IF=13.801]　Duan Xiaojiang. et al. First-in-human study of the radioligand 68Ga-N188 targeting nectin-4 for PET/CT imaging of advanced urothelial carcinoma. CLIN CANCER RES. 2023 Apr;:　　WB　;　　Human.　　 PubMed:37093191 [IF=10.753]　Zhuoying Hu. et al. MitomiR-504 alleviates the copper-induced mitochondria-mediated apoptosis by suppressing Bak1 expression in porcine jejunal epithelial cells. SCI TOTAL ENVIRON. 2023 Feb;858:160157　　WB　;　　Pig.　　 PubMed:36379340 [IF=8.811]　Yamashita Koichiro. et al. Mitotic phosphorylation of Pex14p regulates peroxisomal import machinery. J Cell Biol. 2020 Oct;219(10):e202001003.　　WB　;　　Human.　　 PubMed:32854114 [IF=7.655]　Liu Jincheng. et al. WTAP-Mediated m6A RNA Methylation Regulates the Differentiation of Bone Marrow Mesenchymal Stem Cells via the miR-29b-3p/HDAC4 Axis. Stem Cells Translational Medicine. 2023 Apr;:　　WB　;　　Mouse.　　 PubMed:37010483 [IF=6.684]　Lijin Guo. et al. Whole Transcriptome Analysis Reveals a Potential Regulatory Mechanism of LncRNA-FNIP2/miR-24-3p/FNIP2 Axis in Chicken Adipogenesis. Front Cell Dev Biol. 2021; 9: 653798　　WB　;　　Chicken.　　 PubMed:34249911 [IF=6.656]　Junxuan Wu. et al. Cathepsin B/HSP70 complex induced by Ilexsaponin I suppresses NLRP3 inflammasome activation in myocardial ischemia/reperfusion injury. PHYTOMEDICINE. 2022 Jul;:154358　　WB　;　　Rat.　　 PubMed:35952578 [IF=6.656]　Ying Yang. et al. Formononetin improves cardiac function and depressive behaviours in myocardial infarction with depression by targeting GSK-3β to regulate macrophage/microglial polarization. PHYTOMEDICINE. 2022 Dec;:154602　　WB　;　　Mouse.　　 PubMed:36610138 [IF=6.208]　Liangliang Zhang. et al. Inhibition of EZH2 Causes Retrotransposon Derepression and Immune Activation in Porcine Lung Alveolar Macrophages. INT J MOL SCI. 2023 Jan;24(3):2394　　WB　;　　Porcine.　　 PubMed:36768720 [IF=6.208]　Jiawei Mo. et al. Construction and Analysis of Disuse Atrophy Model of the Gastrocnemius Muscle in Chicken. INT J MOL SCI. 2022 Jan;23(13):6892　　WB　;　　Chicken.　　 PubMed:35805900 [IF=6.208]　Rushuang Xu. et al. Plectin Downregulation Inhibits Migration and Suppresses Epithelial Mesenchymal Transformation of Hepatocellular Carcinoma Cells via ERK1/2 Signaling. INT J MOL SCI. 2023 Jan;24(1):73　　WB　;　　Human.　　 PubMed:36613521 [IF=6.208]　Meng Li. et al. CircRNA Profiling of Skeletal Muscle in Two Pig Breeds Reveals CircIGF1R Regulates Myoblast Differentiation via miR-16. INT J MOL SCI. 2023 Jan;24(4):3779　　WB　;　　Pig.　　 PubMed:36835196 [IF=6.208]　Jinqi Zhang. et al. Study on the Mechanism of MC5R Participating in Energy Metabolism of Goose Liver. INT J MOL SCI. 2023 Jan;24(10):8648　　WB　;　　Geese.　　 PubMed:37239994 [IF=6.175]　Li Kan. et al. m6A demethylase FTO regulate CTNNB1 to promote adipogenesis of chicken preadipocyte. Journal of Animal Science and Biotechnology. 2022 Dec;13(1):1-15　　WB　;　　Chicken.　　 PubMed:36461116 [IF=6.116]　Ping Gong. et al. p47 phox deficiency improves cognitive impairment and attenuates tau hyperphosphorylation in mouse models of AD. Alzheimers Res Ther. 2020 Dec;12(1):1-18　　WB　;　　Mouse.　　 PubMed:33183342 [IF=6.055]　Juan Feng. et al. Melatonin prevents cyclophosphamide-induced primordial follicle loss by inhibiting ovarian granulosa cell apoptosis and maintaining AMH expression. FRONT ENDOCRINOL. 2022; 13: 895095　　WB　;　　Mouse.　　 PubMed:35992124 [IF=5.923]　Yansiwei Cheng. et al. Simulated Microgravity Inhibits Rodent Dermal Fibroblastic Differentiation of Mesenchymal Stem Cells by Suppressing ERK/β-Catenin Signaling Pathway. Int J Mol Sci. 2021 Jan;22(19):10702　　WB　;　　Rat.　　 PubMed:34639043 [IF=5.895]　Xixi Wang. et al. Cannabidiol Alleviates Perfluorooctanesulfonic Acid-Induced Cardiomyocyte Apoptosis by Maintaining Mitochondrial Dynamic Balance and Energy Metabolic Homeostasis. J AGR FOOD CHEM. 2023;XXXX(XXX):XXX-XXX　　WB　;　　Mouse,Rat.　　 PubMed:37010249 [IF=5.652]　You Li. et al. LncRNA PVT1 is a novel mediator promoting the angiogenesis response associated with collateral artery formation. INT J BIOCHEM CELL B. 2022 Aug;:106294　　WB　;　　Human.　　 PubMed:36041701 [IF=5.652]　Haijun Sun. et al. WD Repeat Domain 43 promotes malignant progression of non-small cell lung cancer by regulating CDK2. INT J BIOCHEM CELL B. 2022 Aug;:106293　　WB　;　　Human.　　 PubMed:10.1016/j.biocel.2022.106293 [IF=5.572]　Baoxin Qiao. et al. Curcumin attenuates AFB1-induced duck liver injury by inhibiting oxidative stress and lysosomal damage. FOOD CHEM TOXICOL. 2022 Dec;:113593　　WB　;　　Duck.　　 PubMed:36596445 [IF=5.201]　Chao X et al. miR-429-3p/LPIN1 Axis Promotes Chicken Abdominal Fat Deposition via PPARγ PathwayFront Cell Dev Biol.2020 Dec 21;8:595637.　　WB　;　　chicken.　　 PubMed:33425901 [IF=5.195]　Shuang Chen. et al. Melatonin activates the Mst1-Nrf2 signaling to alleviate cardiac hypertrophy in pulmonary arterial hypertension. EUR J PHARMACOL. 2022 Sep;:175262　　WB　;　　Rat.　　 PubMed:36100129 [IF=4.932]　Zhe Song. et al. Isoandrographolide inhibits NLRP3 inflammasome activation and attenuates silicosis in mice. Int Immunopharmacol. 2022 Apr;105:108539　　WB　;　　Human.　　 PubMed:35063750 [IF=4.932]　Xueli Bai. et al. Regulatory role of methionine enkephalin in myeloid-derived suppressor cells and macrophages in human cutaneous squamous cell carcinoma. Int Immunopharmacol. 2021 Oct;99:107996　　WB　;　　Human.　　 PubMed:34311187 [IF=4.667]　Guo-Dong Wu. et al. Cordyceps Improves Obesity and its Related Inflammation via Modulation of Enterococcus cecorum Abundance and Bile Acid Metabolism. 2022 Mar 10　　WB　;　　Mouse.　　 PubMed:35282803 [IF=4.432]　Hongxiao Yang. et al. Hydrogen Attenuates Thyroid Hormone-Induced Cardiac Hypertrophy in Rats by regulating angiotensin II type 1 receptor and NADPH oxidase 2 mediated oxidative stress. Eur J Pharmacol. 2022 May;922:174917　　WB　;　　Rat.　　 PubMed:35341785 [IF=4.155]　Quanwei Li. et al. Toxicological mechanism of large amount of copper supplementation: Effects on endoplasmic reticulum stress and mitochondria-mediated apoptosis by Nrf2/HO-1 pathway-induced oxidative stress in the porcine myocardium. J Inorg Biochem. 2022 May;230:111750　　WB　;　　Pig.　　 PubMed:35151098 [IF=4.145]　Kunlin Wu. et al. Histone deacetylase inhibitor panobinostat in combination with rapamycin confers enhanced efficacy against triple-negative breast cancer. EXP CELL RES. 2022 Sep;:113362　　WB　;　　Human.　　 PubMed:36152730 [IF=4.115]　Yaodong Zhou. et al. Association of MicroRNA-21 with p53 at Mutant Sites R175H and R248Q, Clinicopathological Features, and Prognosis of NSCLC. Mol Ther-Oncolytics. 2020 Dec;19:208　　WB　;　　Human.　　 PubMed:33251333 [IF=4.109]　He Bai. et al. Inhibition of the BNIP3/NIX-dependent mitophagy aggravates copper-induced mitochondrial dysfunction in duck renal tubular epithelial cells. ENVIRON TOXICOL. 2022 Nov;:　　WB　;　　Duck.　　 PubMed:36378575
產品類型	內參抗體
研究領域	腫瘤　　細胞生物　　免疫學　　神經生物學　　新陳代謝
抗體來源	Mouse
克隆類型	Monoclonal
克　隆　號	4F8
交叉反應	Human,Mouse,Rat,Pig,Hamster (predicted: Chicken,Dog,Rabbit,Sheep,Monkey)
產品應用	WB=1:10000-1:100000, IHC-P=1:100-500, ICC=1:100 not yet tested in other applications. optimal dilutions/concentrations should be determined by the end user.
理論分子量	38kDa
細胞定位	細胞核　細胞漿　細胞膜
性　　　　狀	Liquid
濃　　　　度	1mg/ml
免　疫　原	Recombinded Human GAPDH
亞　　　　型	IgG
純化方法	affinity purified by Protein G
緩　沖　液	0.01M TBS(pH7.4) with 1% BSA, 0.03% Proclin300 and 50% Glycerol.
保存條件	Shipped at 4℃. Store at -20 °C for one year. Avoid repeated freeze/thaw cycles.
注意事項	This product as supplied is intended for research use only, not for use in human, therapeutic or diagnostic applications.
PubMed	PubMed
產品介紹	Loading Control Glyceraldehyde 3 phosphate dehydrogenase (GAPDH) is well known as one of the key enzymes involved in glycolysis. As well as functioning as a glycolytic enzyme in cytoplasm, recent evidence suggests that mammalian GAPDH is also involved in a great number of intracellular proceses such as membrane fusion, microtubule bundling, phosphotransferase activity, nuclear RNA export, DNA replication, and DNA repair. During the last decade a lot of data appeared concerning the role of GAPDH in different pathologies including prostate cancer progression, programmed neuronal cell death, age related neuronal diseases, such as Alzheimer's and Huntington's disease. GAPDH is expressed in all cells. It is constitutively expressed in almost all tissues at high levels. There are however some physiological factors such as hypoxia and diabetes that increase GAPDH expression in certain cell types. GAPDH molecule is composed of four 36kDa subunits. Function: 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. 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. Subunit: Homotetramer. Interacts with TPPP; the interaction is direct. Interacts (when S-nitrosylated) with SIAH1; leading to nuclear translocation. Interacts with RILPL1/GOSPEL, leading to prevent the interaction between GAPDH and SIAH1 and prevent nuclear translocation. Interacts with EIF1AD, USP25, PRKCI and WARS. Subcellular Location: Cytoplasm, cytosol. Nucleus. Cytoplasm, perinuclear region. Membrane. Note=Translocates to the nucleus following S-nitrosylation and interaction with SIAH1, which contains a nuclear localization signal. Postnuclear and Perinuclear regions. Post-translational modifications: S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus. ISGylated (Probable). Sulfhydration at Cys-152 increases catalytic activity. Similarity: Belongs to the glyceraldehyde-3-phosphate dehydrogenase family. SWISS: P04406 Gene ID: 2597 Database links: Entrez Gene: 374193　Chicken Entrez Gene: 2597　Human Entrez Gene: 100042025　Mouse Entrez Gene: 14433　Mouse Entrez Gene: 24383　Rat Entrez Gene: 685186　Rat Entrez Gene: 317743　Zebrafish Omim: 138400　Human SwissProt: P00356　Chicken SwissProt: P04406　Human SwissProt: P16858　Mouse SwissProt: P04797　Rat SwissProt: Q5XJ10　Zebrafish GAPDH蛋白幾乎在所有組織中都高水平表達，廣泛用作Western blot蛋白質標準化的內參,是很好的內參抗體。 GAPDH 作為管家基因在同種細胞或者組織中的蛋白質表達量一般是恒定的。在實驗中，可能存在總蛋白濃度測定不準確；或者蛋白質樣品在電泳前上樣時產生的樣品間的操作誤差；這些誤差需要通過測定每個樣品中實際轉到膜上的GAPDH的含量來進行校正，所以一般的western實驗都需要進行內參設置。具體校正的方法就是將每個樣品測得的目的蛋白含量與本樣品的GAPDH含量相除，得到每個樣品目的蛋白的相對含量。然后才進行樣品與樣品之間的比較。甘油醛-3-磷酸脫氫酶（Glyceraldehyde 3 phosphate dehydrogenase，GAPDH）是糖酵解（glycolysis）過程中的關鍵酶。除了在胞質中作為糖酵解的酶以外，有證據表明哺乳動物細胞中的GAPDH參與了多種胞內生化過程，包括膜融合（membrane fusion）、微管成束（microtubule bundling）、磷酸轉移酶（phosphotransferase）激活、核內RNA出核、DNA復制與DNA修復。一些生理因素，諸如低氧（hypoxia）和尿糖（diabetes），可以增加GAPDH在特定細胞中的表達。GAPDH存在于幾乎所有的組織中，以高水平持續表達。 GAPDH（甘油醛-3-磷酸脫氫酶）是參與糖酵解的一種關鍵酶，由4個30－40kDa的亞基組成.
產品圖片	Sample: Lane1: Skin (Mouse) Lysate at 40 ug Lane2: Testis (Mouse) Lysate at 40 ug Lane3: Adrenal gland (Mouse) Lysate at 40 ug Primary: Anti-GAPDH (bsm-33033M) at 1/1 000 dilution Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution Predicted band size: 38 kDa Observed band size: 38 kDa Sample: Lane 1: Rat Cerebrum tissue lysates Lane 2: Rat Heart tissue lysates Lane 3: Human A549 cell lysates Lane 4: Human Jurkat cell lysates Lane 5: Human Huvec cell lysates Lane 6: Human Hela cell lysates Lane 7: Human U2os cell lysates Lane 8: Human HepG2 cell lysates Primary: Anti- GAPDH (bsm-33033M) at 1/50000 dilution Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution Predicted band size: 38 kDa Observed band size: 38 kDa Sample: Lane 1~4：Cerebrum (Rat) Lysate at 40 ug Lane 5~8：Hela (Human) Lysate at 40 ug Primary: Anti-GAPDH (bsm-33033M) at 1/2000~1/20000 dilution Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution Predicted band size: 38 kDa Observed band size: 38 kDa Sample: Lane 1: Human HEK293 cell lysates Lane 2: Human Hela cell lysates Lane 3: Human SH-SY5Y cell lysates Lane 4: Mouse NIH/3T3 cell lysates Lane 5: Hamster CHO cell lysates Lane 6: Rat Brain tissue lysates Primary: Anti-GAPDH (bsm-33033M) at 1/200000 dilution Secondary: IRDye800CW Goat Anti-Mouse IgG at 1/20000 dilution Predicted band size: 38 kDa Observed band size: 38 kDa Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GAPDH-Loading Control) Monoclonal Antibody, Unconjugated (ascites of bsm-33033M-4E8) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining. Paraformaldehyde-fixed, paraffin embedded (rat spleen); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GAPDH-Loading Control) Monoclonal Antibody, Unconjugated (ascites of bsm-33033M-4E8) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining. Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GAPDH-Loading Control) Monoclonal Antibody, Unconjugated (ascites of bsm-33033M-4E8) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining. Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GAPDH-Loading Control) Monoclonal Antibody, Unconjugated (ascites of bsm-33033M-4E8) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining. Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GAPDH-Loading Control) Monoclonal Antibody, Unconjugated (ascites of bsm-33033M-4E8) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining. Paraformaldehyde-fixed, paraffin embedded (rat spleen); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GAPDH-Loading Control) Monoclonal Antibody, Unconjugated (ascites of bsm-33033M-4E8) at 1:2000 overnight at 4°C, followed by operating according to SP Kit(Mouse) (sp-0024) instructions and DAB staining. Paraformaldehyde-fixed, paraffin embedded (human gastric); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GAPDH-Loading Control) Monoclonal Antibody, Unconjugated (bsm-33033M ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (human colon carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GAPDH-Loading Control) Monoclonal Antibody, Unconjugated (bsm-33033M ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GAPDH-Loading Control) Monoclonal Antibody, Unconjugated (bsm-33033M ) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (human gastric carcinoma); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GAPDH-Loading Control) Monoclonal Antibody, Unconjugated (bsm-33033M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (human gastric); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GAPDH-Loading Control) Monoclonal Antibody, Unconjugated (bsm-33033M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (Human kidney); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GAPDH-Loading Control) Monoclonal Antibody, Unconjugated (bsm-33033M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (rat brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GAPDH-Loading Control) Monoclonal Antibody, Unconjugated (bsm-33033M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining. Paraformaldehyde-fixed, paraffin embedded (mouse brain); Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with (GAPDH-Loading Control) Monoclonal Antibody, Unconjugated (bsm-33033M) at 1:200 overnight at 4°C, followed by operating according to SP Kit(Mouse)(sp-0024) instructionsand DAB staining. Tissue/cell:Hela cell; 4% Paraformaldehyde-fixed; Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum, C-0005) at 37°C for 20 min; Antibody incubation with (GAPDH-Loading Control) monoclonal Antibody, Unconjugated (bsm-33033M) 1:100, 90 minutes at 37°C; followed by a CY3 conjugated Goat Anti-Mouse IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.