Product Info

Source:
Mouse
Application:
WB 1:3000-1:10000, IF/ICC: 1:200, IP 1:200, IHC 1:50-1:200
*The optimal dilutions should be determined by the end user.
*Tips:

WB: For western blot detection of denatured protein samples. IHC: For immunohistochemical detection of paraffin sections (IHC-p) or frozen sections (IHC-f) of tissue samples. IF/ICC: For immunofluorescence detection of cell samples. ELISA(peptide): For ELISA detection of antigenic peptide.

Reactivity:
All
Clonality:
Monoclonal [T177]
RRID:
AB_2839412
Cite Format: Affinity Biosciences Cat# T0003, RRID:AB_2839412.
Conjugate:
Unconjugated.
Purification:
Affinity-chromatography.
Storage:
Mouse IgG1 in phosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, 150mM NaCl, 0.02% sodium azide and 50% glycerol. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

DDDDK epitope tag; DDDK; ddk; DYKDDDDK; DYKDDDDK epitope tag; DYKDDDDK tag; ECS epitope tag; ECS tag; Enterokinase Cleavage Site epitope tag; Enterokinase Cleavage Site tag; FLAG; FLAG tag antibody

Immunogens

Immunogen:

A synthetic peptide DYKDDDDK coupled to KLH.

Description:
FLAG tag Mouse mAb is part of the series of Tag antibodies, the excellent quality in the research. FLAG tag antibody is a highly sensitive and affinity PAB applicable to FLAG-tagged fusion protein detection. FLAG tag antibody can detect FLAG tags in internal, C-terminal, or N-terminal recombinant proteins.

References

1). Liu H et al. Psychologic Stress Drives Progression of Malignant Tumors via DRD2/HIF1α Signaling. Cancer Res 2021 Oct 15;81(20):5353-5365. (PubMed: 34321238) [IF=13.312]

Application: WB    Species: mouse    Sample:

Fig. 4 |DRD2 overexpression inhibited the ubiquitin-dependent degradation of HIF1α. N, Effect of DRD2 on the content of ubiquitin binding to HIF1α. DRD2-overexpressing cells treated with MG132, and cellular extracts were prepared for co-IP assay with anti–HIF1α antibody, followed by Western blotting detection of ubiquitin. siRNA, small interfering RNA. ns, nonsignificant; *, P < 0.05; **, P < 0.01

2). Liu W et al. Micheliolide Ameliorates Diabetic Kidney Disease by Inhibiting Mtdh-mediated Renal Inflammation in Type 2 Diabetic db/db Mice. Pharmacol Res 2019 Oct 24:104506 (PubMed: 31669149) [IF=10.334]

3). Ye F et al. circFBXW7 Inhibits Malignant Progression by Sponging miR-197-3p and Encoding a 185-aa Protein in Triple-Negative Breast Cancer. Mol Ther Nucleic Acids 2019 Aug 14;18:88-98 (PubMed: 31536884) [IF=10.183]

Application: WB    Species: human    Sample: BT549 cells

Figure 6.| FBXW7-185aa, Encoded by circFBXW7, Inhibited the Proliferation and Metastasis of TNBC Cells through Downregulating c-Myc Expression.s. (H)FBXW7-185aa-FLAG-overexpressing BT549 cells were cotransfected with sh-FBXW7 or USP28 vectors. The FBXW7-185aaFLAG, FBXW7, USP28, and c-Myc expression levels were quantified by western blot analysis.

4). Matsuoka TA et al. Mafa Enables Pdx1 to Effectively Convert Pancreatic Islet Progenitors and Committed Islet α-Cells Into β-Cells In Vivo. Diabetes 2017 May;66(5):1293-1300 (PubMed: 28223284) [IF=9.337]

Application: IF/ICC    Species: mouse    Sample: islet cells

Supplemental Figure 1| Pdx1 flag is expressed in most P0.5 and 6W Ngn3-Cre;CAT-Pdx1 islet cells. Flag staining (red) was observed in most of Ngn3-Cre;CAT-Pdx1 insulin + (blue) and glucagon + (green) cells at P0.5 and 6W; the Crepenetrance rate was a maintained at approximately 95%. Scale bars: 50 mm.

5). Ma Y et al. SIRT5-mediated SDHA desuccinylation promotes clear cell renal cell carcinoma tumorigenesis. Free Radic Biol Med 2019 Apr;134:458-467 (PubMed: 30703481) [IF=8.101]

Application: WB    Species: human    Sample: HEK293A cells

Fig. 6.| SIRT5 promotes ccRCC cell proliferation by interacting with and desuccinylating SDHA. (A-B) ACHN cells were transfected with scramble shRNA or SIRT5 shRNA plasmid, SDHA was precipitated and its succinylation level was probed by anti-pan-succinylation antibody. SIRT5 silencing led to hypersuccinylation of SDHA in ccRCC cells and increased activity of SDHA. (C-E) SIRT5 silencing decreased cellular NADPH/NADP+and GSH/GSSG ratio and increased cellular ROS level. (F) Co-IP of SIRT5 and FLAG-tagged WT SDHA in HEK293A cells revealed SIRT5 coimmunoprecipitated with SDHA. (G-J) SIRT5 silencing repressed ACHN and 769-P cell proliferation and colony formation. Methods followed Fig. 5G-J.

6). Wang Y et al. FGFR2 Mutation p.Cys342Arg Enhances Mitochondrial Metabolism-Mediated Osteogenesis via FGF/FGFR-AMPK-Erk1/2 Axis in Crouzon Syndrome. Cells 2022 Oct 5;11(19):3129. (PubMed: 36231091) [IF=6.600]

7). Zou J et al. WWP2 drives the progression of gastric cancer by facilitating the ubiquitination and degradation of LATS1 protein. Cell Commun Signal 2023 Feb 17;21(1):38. (PubMed: 36803368) [IF=5.712]

8). Li Y et al. Neuropilin-2 Signaling Modulates Mossy Fiber Sprouting by Regulating Axon Collateral Formation Through CRMP2 in a Rat Model of Epilepsy. Mol Neurobiol 2022 Aug 31. (PubMed: 36044155) [IF=5.682]

Application: WB    Species: Rat    Sample: hippocampal

Fig. 5Npn-2 controls axon collateral formation. A Npn-2 knockdown by shNpn-2 validated in primary neuron cultures by western blot using an antibody for rat Npn-2. B Quantitation of Npn-2 in A, n = 3, ***P = 0.0003. C hNpn-2 expression validated by western blot. Expression of hNpn-2 in neurons was detected by western blot using an anti-Flag antibody, showing that the expression of human full-length Npn-2 cannot be affected by the rat shNpn-2 used. D Quantitation of Npn-2 in C, n = 3, P > 0.05. E Neonatal rat primary hippocampal neurons were transfected with shCtrl, shNpn-2, or shNpn-2 plus hNpn-2 plasmids at DIV 0 and treated with either control medium or Sema3F (5 nM) at 48 h after transfection for 24 h. Axons and their collaterals were detected by immunofluorescent staining with GFP and Tau-1. Arrows indicate main axons and stars indicate axon collaterals. Scale bar, 40 μm. F, G Quantitation of axon collateral number and length in E. Sema3F treatment reduced the number and length of axon collaterals in neurons transfected with shCtrl but not in neurons with shNpn-2 transfection, which was rescued by hNpn-2 co-transfection. *P < 0.05, **P < 0.01. H Quantitation of main axon length in E. Main axon elongation in neurons transfected with shCtrl was inhibited upon Sema3F treatment, but not in neurons with shNpn-2 transfection, which was rescued by hNpn-2 expression. **P < 0.01, ***P < 0.001. One-way ANOVA, post hoc Tukey test. Error bars represent SEM

9). Sun L et al. Overexpression of lncRNA-Gm2044 in spermatogonia impairs spermatogenesis in partial seminiferous tubules. Int J Mol Med 2022 Feb;49(2):17. (PubMed: 34935058) [IF=5.314]

10). Wang C et al. Gankyrin activates the hedgehog signalling to drive metastasis in osteosarcoma. J Cell Mol Med 2021 Jun 5. (PubMed: 34089292) [IF=5.295]

Application: IHC    Species: Human    Sample: U2OS and MG63 cells

FIGURE 1 Gankyrin promotes migration and invasion in OS in vitro. A, Induction of gankyrin in U2OS and MG63 cells was confirmed by Western blot analysis. B, Knockdown of gankyrin in U2OS and MG63 cells was confirmed via Western blot analysis. C, Induction of gankyrin significantly increased OS cell migration ability, as shown by wound healing assays. *P < .05; **P < .01. D, Up‐regulated gankyrin remarkably promoted OS cell invasion, as shown by Transwell invasion assays. *P < .05. E, Knockdown of gankyrin reduced OS cell migration ability. **P < .01. F, Knockdown of gankyrin inhibited OS cell invasion. **P < .01; ***P < .001

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Affinity Biosciences tests all products strictly. Citations are provided as a resource for additional applications that have not been validated by Affinity Biosciences. Please choose the appropriate format for each application and consult Materials and Methods sections for additional details about the use of any product in these publications.

For Research Use Only.
Not for use in diagnostic or therapeutic procedures. Not for resale. Not for distribution without written consent. Affinity Biosciences will not be held responsible for patent infringement or other violations that may occur with the use of our products. Affinity Biosciences, Affinity Biosciences Logo and all other trademarks are the property of Affinity Biosciences LTD.