Anti-GAPDH 抗体 [EPR6256] - Loading Control (ab128915)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR6256] to GAPDH - Loading Control
- Suitable for: WB, IP, ICC/IF, IHC-P, Flow Cyt (Intra)
- Reacts with: Human, African green monkey
Related conjugates and formulations
製品の概要
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製品名
Anti-GAPDH antibody [EPR6256] - Loading Control
GAPDH 一次抗体 製品一覧 -
製品の詳細
Rabbit monoclonal [EPR6256] to GAPDH - Loading Control -
由来種
Rabbit -
特異性
The African green monkey recommendation is based on the WB results. We do not guarantee IHC-P for African green monkey.
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アプリケーション
適用あり: WB, IP, ICC/IF, IHC-P, Flow Cyt (Intra)more details -
種交差性
交差種: Human, African green monkey -
免疫原
Synthetic peptide within Human GAPDH aa 250 to the C-terminus. The exact sequence is proprietary.
Database link: P04406 -
ポジティブ・コントロール
- WB: 293T, HeLa, HepG2, HUVEC, MCF7 and SH-SY5Y cell lysates. ICC/IF: HeLa and MCF7 cells. IHC-P: Human pancreas tissue, Human bladder carcinoma. Flow Cyt (intra): HeLa cells. IP: HeLa cell lysate.
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特記事項
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Mouse: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
製品の特性
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製品の状態
Liquid -
保存方法
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle. Stable for 12 months at -20°C. -
バッファー
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA -
Concentration information loading...
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精製度
Protein A purified -
ポリ/モノ
モノクローナル -
クローン名
EPR6256 -
アイソタイプ
IgG -
研究分野
関連製品
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Alternative Versions
- HRP Anti-GAPDH antibody [EPR6256] - Loading Control (ab185059)
- Anti-GAPDH antibody [EPR6256] - BSA and Azide free (ab186930)
- Biotin Anti-GAPDH antibody [EPR6256] - Loading Control (ab195904)
- Alexa Fluor® 647 Anti-GAPDH antibody [EPR6256] (ab215227)
- PerCP Anti-GAPDH antibody [EPR6256] (ab216377)
- APC Anti-GAPDH antibody [EPR6256] (ab221270)
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
アプリケーション
The Abpromise guarantee
Abpromise保証は、 次のテスト済みアプリケーションにおけるab128915の使用に適用されます
アプリケーションノートには、推奨の開始希釈率がありますが、適切な希釈率につきましてはご検討ください。
アプリケーション | Abreviews | 特記事項 |
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WB | (9) |
1/10000 - 1/50000. Detects a band of approximately 35 kDa (predicted molecular weight: 36 kDa).
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IP |
1/10 - 1/100.
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ICC/IF | (1) |
1/250 - 1/500.
2.0 µg/ml |
IHC-P |
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
See IHC antigen retrieval protocols. For unpurified use at 1/250. The African green monkey recommendation is based on the WB results. We do not guarantee IHC-P for African green monkey. |
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Flow Cyt (Intra) |
1/20.
For unpurified use at 1/100 - 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
特記事項 |
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WB
1/10000 - 1/50000. Detects a band of approximately 35 kDa (predicted molecular weight: 36 kDa). |
IP
1/10 - 1/100. |
ICC/IF
1/250 - 1/500. 2.0 µg/ml |
IHC-P
1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. See IHC antigen retrieval protocols. For unpurified use at 1/250. The African green monkey recommendation is based on the WB results. We do not guarantee IHC-P for African green monkey. |
Flow Cyt (Intra)
1/20. For unpurified use at 1/100 - 1/1000. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
ターゲット情報
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機能
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. -
パスウェイ
Carbohydrate degradation; glycolysis; pyruvate from D-glyceraldehyde 3-phosphate: step 1/5. -
配列類似性
Belongs to the glyceraldehyde-3-phosphate dehydrogenase family. -
翻訳後修飾
S-nitrosylation of Cys-152 leads to interaction with SIAH1, followed by translocation to the nucleus.
ISGylated. -
細胞内局在
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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参照データベース
- Entrez Gene: 2597 Human
- Omim: 138400 Human
- SwissProt: P04406 Human
- Unigene: 544577 Human
- Unigene: 592355 Human
- Unigene: 598320 Human
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別名
- 38 kDa BFA-dependent ADP-ribosylation substrate antibody
- aging associated gene 9 protein antibody
- Aging-associated gene 9 protein antibody
see all
画像
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Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915) at 1/20000 dilution (Purified) + COS-1 (African green monkey kidney fibroblast-like) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa -
Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915) at 1/20000 dilution (Purified) + HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 15 µg
Secondary
Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 36 kDa
Observed band size: 36 kDa -
All lanes : Anti-GAPDH antibody [EPR6256] - Loading Control (ab128915) at 1/10000 dilution (unpurified)
Lane 1 : HEK-293T (human epithelial cell line from embryonic kidney transformed with large T antigen) cell lysate
Lane 2 : HeLa (human epithelial cell line from cervix adenocarcinoma) cell lysate
Lane 3 : HepG2 (human liver hepatocellular carcinoma cell line) cell lysate
Lane 4 : HUVEC (human umbilical vein endothelial cell line) cell lysate
Lane 5 : MCF7 (human breast adenocarcinoma cell line) cell lysate
Lane 6 : SH-SY5Y (human neuroblastoma cell line from bone marrow) cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP labelled Goat anti-Rabbit IgG at 1/2000 dilution
Predicted band size: 36 kDa
Observed band size: 35 kDa why is the actual band size different from the predicted?Secondary antibody - anti-rabbit HRP (ab6721)
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human bladder carcinoma tissue sections labeling GAPDH with purified ab128915 at 1/2000 dilution (0.06 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
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Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling GAPDH with purified ab128915 at 1/250 dilution (0.4 µg/ml). Cells were fixed in 100% Methanol. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 (2.5 µg/ml) dilution. Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1/1000 (2 µg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.
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Intracellular Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling GAPDH with purified ab128915 at 1/20 dilution (10µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluorr® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue).
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ab128915 (purified) at 1/20 dilution (0.5ug) immunoprecipitating GAPDH in HeLa whole cell lysates.
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates 10ug
Lane 2 (+): ab128915 & HeLa whole cell lysates
Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab128915 in HeLa whole cell lysates
For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM/TBST. -
IHC image of ab128915 (unpurified) staining GAPDH in human pancreas* formalin fixed paraffin embedded tissue sections, performed on a Leica Bond. 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 ab128915, 1:250 dilution, for 15 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. No primary antibody was used in the secondary only control (shown on the inset).
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre -
ab128915 (unpurified) staining GAPDH in HeLa (human epithelial cell line from cervix adenocarcinoma) cells. The cells were fixed with 100% methanol (5min) and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab128915 at 2μg/ml and ab7291 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an goat anti-rabbit AlexaFluor®488 (ab150081) at 2 μg/ml (shown in green) and goat anti-mouse AlexaFluor®594 (ab150120) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled 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.
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Overlay histogram showing HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained with ab128915 (unpurified) (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab128915, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was goat anti-rabbit Alexa Fluor® 488 (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
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ab128915 (unpurified) staining GAPDH in human HeLa (human epithelial cell line from cervix adenocarcinoma) cells by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde and permeabilized with 0.5% Triton X-100 in PBS. Samples were incubated with primary antibody (1/500 in PBS) for 1 hour at 22°C. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG polyclonal (1/200) was used as the secondary antibody. Counter stained with DAPI.
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ab128915 (unpurified), at 1/250, staining GAPDH in MCF7 (human breast adenocarcinoma cell line) cells by immunofluorescence.
プロトコール
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (246)
ab128915 は 246 報の論文で使用されています。
- Wang L et al. Circ_0000520 interacts with miR-512-5p to upregulate KIAA0100 to promote malignant behaviors in lung cancer. Histol Histopathol 38:73-89 (2023). PubMed: 35866672
- Bons J et al. Data-independent acquisition and quantification of extracellular matrix from human lung in chronic inflammation-associated carcinomas. Proteomics 23:e2200021 (2023). PubMed: 36228107
- Martínez-Sabadell A et al. The target antigen determines the mechanism of acquired resistance to T cell-based therapies. Cell Rep 41:111430 (2022). PubMed: 36261015
- Steffens Reinhardt L et al. Alterations in the p53 isoform ratio govern breast cancer cell fate in response to DNA damage. Cell Death Dis 13:907 (2022). PubMed: 36307393
- Li J et al. MiR-137 targets and regulates E2F7 to suppress progression of glioma cells. Folia Neuropathol 60:346-354 (2022). PubMed: 36382488