Anti-GAPDH抗体[mAbcam 9484] - Loading Control (ab9484)
Key features and details
- Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control
- Suitable for: WB, IHC-P
- Reacts with: Mouse, Rat, Chicken, Cow, Human, Pig, Xenopus laevis, Chinese hamster
- Isotype: IgG2b
Related conjugates and formulations
概述
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产品名称
Anti-GAPDH抗体[mAbcam 9484] - Loading Control
参阅全部 GAPDH 一抗 -
描述
小鼠单克隆抗体[mAbcam 9484] to GAPDH - Loading Control -
宿主
Mouse -
特异性
In western blot, this product typically gives a lower signal in rat lysates compared to human and mouse lysates.
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经测试应用
适用于: WB, IHC-Pmore details -
种属反应性
与反应: Mouse, Rat, Chicken, Cow, Human, Pig, Xenopus laevis, Chinese hamster
预测可用于: Rabbit, Dog, Cynomolgus monkey -
免疫原
Full length native protein (purified). This information is proprietary to Abcam and/or its suppliers.
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阳性对照
- WB: A431, Jurkat, HEK293, HeLa, NIH 3T3, PC-12, EBTr and CHO cell lysates; Rat brain, chicken and pig liver and Xenopus laevis embryo tissue lysates. IHC-P: Human liver tissue.
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常规说明
For Western blotting, do not use milk for blocking. Our labs have extensively tested the blocking conditions for this antibody and recommend using 5% BSA for 1 hour. The comparison data is shown in the images section.
This antibody clone [mAbcam 9484] is manufactured by Abcam.
If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.
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 this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, 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: 0.02% Sodium azide
Constituents: PBS, 6.97% L-Arginine -
Concentration information loading...
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纯度
IgG fraction -
克隆
单克隆 -
克隆编号
mAbcam 9484 -
骨髓瘤
Sp2/0-Ag14 -
同种型
IgG2b -
轻链类型
kappa -
研究领域
相关产品
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Alternative Versions
- Anti-GAPDH antibody [mAbcam 9484] - BSA and Azide free (ab174646)
- Alexa Fluor® 680 Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab184095)
- Alexa Fluor® 790 Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab184578)
- Alexa Fluor® 647 Anti-GAPDH antibody [mAbcam 9484] (ab196697)
- HRP Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9482)
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Compatible Secondaries
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Isotype control
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Recombinant Protein
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Related Products
应用
应用 | Ab评论 | 说明 |
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WB | (53) |
Use a concentration of 0.1 - 1 µg/ml. Predicted molecular weight: 36 kDa.
Do not block with milk. Block with 5% BSA for 1 hour. Our labs have thoroughly investigated the blocking conditions for this antibody. We found that milk significantly decreases the signal and is therefore not a suitable blocking agent for this antibody (see images). |
IHC-P |
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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说明 |
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WB
Use a concentration of 0.1 - 1 µg/ml. Predicted molecular weight: 36 kDa. Do not block with milk. Block with 5% BSA for 1 hour. Our labs have thoroughly investigated the blocking conditions for this antibody. We found that milk significantly decreases the signal and is therefore not a suitable blocking agent for this antibody (see images). |
IHC-P
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. |
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: 374193 Chicken
- Entrez Gene: 281181 Cow
- Entrez Gene: 403755 Dog
- Entrez Gene: 2597 Human
- Entrez Gene: 100042025 Mouse
- Entrez Gene: 14433 Mouse
- Entrez Gene: 396823 Pig
- Entrez Gene: 100009074 Rabbit
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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All lanes : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1 µg/ml
Lane 1 : HeLa Whole Cell Lysate
Lane 2 : NIH 3T3 Whole Cell Lysate
Lane 3 : PC12 Whole Cell Lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 36 kDa
Exposure time: 1 minuteThis blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab9484 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406
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IHC image of GAPDH staining in human liver FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab9484, 5µg/ml, for 8 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
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All lanes : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/5000 dilution
Lane 1 : Hela whole cell (Human)
Lane 2 : 3T3 cell (Mouse)
Lane 3 : Rat brain
Lane 4 : Xenopus laevis embryo
Lane 5 : Chicken Liver
Lane 6 : EBTr cell (Cow)
Lane 7 : CHO cell (Chinese hamster)
Lane 8 : Pig liver
Secondary
All lanes : Rabbit Anti-Mouse IgG H&L (HRP) (ab6728) at 1/5000 dilution
Performed under reducing conditions.
Predicted band size: 36 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?
Exposure time: 10 secondsThe membrane was blocked in 5% BSA in TBST for 1 hour, then incubated for 1 hour in primary antibody diluted in TBST.
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Lanes 1-5 : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/1000 dilution (Blocked in 5% milk)
Lanes 6-10 : Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 1/1000 dilution (Blocked in 5% BSA)
Lanes 1 & 6 : HeLa (Human epithelial carcinoma cell line) Nuclear Lysate
Lanes 2 & 7 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lanes 3 & 8 : A-431 whole cell lysate (ab7909)
Lanes 4 & 9 :Jurkat whole cell lysate (ab7899)
Lanes 5 & 10 : HEK-293 whole cell lysate (ab7902)
Lysates/proteins at 20 µg per lane.
Secondary
Lanes 1-5 : Goat anti-Mouse (HRP conjugated) at 1/5000 dilution
Lanes 6-10 : Goat anti-Mouse (HRP conjugated) at 1/5000 dilution
Predicted band size: 36 kDa
Observed band size: 40 kDa why is the actual band size different from the predicted?The membrane 1-5 was blocked in 5% milk (1 hour). The membrane 6-10 was blocked in 5% BSA (1 hour). Milk is not a suitable blocking agent and significantly decreases the signal on the membrane.
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Anti-GAPDH antibody [mAbcam 9484] - Loading Control (ab9484) at 0.5 µg/ml + HeLa cell lysate
Secondary
Goat Anti-Mouse IgG H&L (HRP) (ab6789) at 1/5000 dilution
Developed using the ECL technique.
Performed under non-reducing conditions.
Predicted band size: 36 kDa
Exposure time: 30 seconds
数据表及文件
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SDS download
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Datasheet download
文献 (875)
ab9484 被引用在 875 文献中.
- Bodnar-Wachtel M et al. Inflammasome-independent NLRP3 function enforces ATM activity in response to genotoxic stress. Life Sci Alliance 6:N/A (2023). PubMed: 36746533
- Sompol P et al. Targeting Astrocyte Signaling Alleviates Cerebrovascular and Synaptic Function Deficits in a Diet-Based Mouse Model of Small Cerebral Vessel Disease. J Neurosci 43:1797-1813 (2023). PubMed: 36746627
- Malik N et al. Dysregulation of Mitochondrial Translation Caused by CBFB Deficiency Cooperates with Mutant PIK3CA and Is a Vulnerability in Breast Cancer. Cancer Res 83:1280-1298 (2023). PubMed: 36799863
- Zou W & Imperiale MJ Regulation of Virus Replication by BK Polyomavirus Small T Antigen. J Virol 97:e0007723 (2023). PubMed: 36916919
- Zhu DD et al. AQP4 Aggravates Cognitive Impairment in Sepsis-Associated Encephalopathy through Inhibiting Nav 1.6-Mediated Astrocyte Autophagy. Adv Sci (Weinh) 10:e2205862 (2023). PubMed: 36922751