Product Info

Source:
Mouse
Application:
WB: 1:3000-1:10000, IF/ICC: 1:200, IP 1:200
*The optimal dilutions should be determined by the end user.
Reactivity:
All
Clonality:
Monoclonal [T80]
Specificity:
HA-Tag Mouse Monoclonal antibody detects endogenous levels of internal, C-terminal, or N-terminal HA-tagged recombinant proteins.
RRID:
AB_2839415
Cite Format: Affinity Biosciences Cat# T0008, RRID:AB_2839415.
Purification:
Affinity-chromatography.
Storage:
Mouse IgG1 in phosphate buffered saline (without Mg2+ and Ca2+), pH 7.4, 150mM NaCl,. Store at -20 °C. Stable for 12 months from date of receipt.
Alias:

Fold/Unfold

ha tag

Immunogens

Immunogen:

A synthetic peptide YPYDVPDYA coupled to KLH.

Description:
N/A

References

1). Liu T et al. The m6A reader YTHDF1 promotes ovarian cancer progression via augmenting EIF3C translation. Nucleic Acids Res 2020 Jan 30 (PubMed: 31996915) [IF=11.501]

Application: WB    Species: Human    Sample: in A2780 or SKOV3 cells

Figure 6. YTHDF1 regulates EIF3C translation in an m6A-dependent manner. (A) Relative RNA level of EIF3C in A2780 and SKOV3 upon YTHDF1 knockdown. (B) Western blot detected the protein level of EIF3C in A2780 and SKOV3 cells upon YTHDF1 knockdown. (C) Gene-specific m6A qPCR validation of m6A levels in A2780 and SKOV3 cells. Primers to m6A negative region of EEF1A as the negative control and primers to m6A postive region of EEF1A as the positive control. (D) YTHDF1 RIP followed by RT-qPCR confirmed the interaction between YTHDF1 and EIF3C mRNA. (E) Schematic representation of wild-type (YTHDF1-wt) and mutant (YTHDF1-mut) YTHDF1 constructs. (F) RIP-derived RNA and protein in A2780 cells were measured by RT-qPCR and western blot, respectively. GAPDH was used as the negative control in western blot assays. (G) Western blot confirmed HAtagged EIF3C expression in A2780 or SKOV3 cells co-transfected with empty vector, wild-type or mutant Flag-tagged YTHDF1 and wild-type or mutant HA-tagged EIF3C. (H) Nascent protein synthesis was detected by HPG incorporation upon YTHDF1 knockdown or EIF3C knockdown in A2780 cells. Scale bar, 100  m. Data are shown as means ± S.D. *P < 0.05, **P < 0.01, ***P < 0.001, NS, not significant.

2). Zhang H et al. Oryza sativa POSITIVE REGULATOR OF IRON DEFICIENCY RESPONSE 2 (OsPRI2) and OsPRI3 are involved in the maintenance of Fe homeostasis. Plant Cell Environ 2020 Jan;43(1):261-274 (PubMed: 31674679) [IF=6.362]

Application: WB    Species: yeast    Sample: yeast cells

FIGURE 1| Interaction of OsHRZ1 with OsPRI2 and OsPRI3.(c) CoIP assay. Total proteins from different combinations with OsHRZ1‐GFP and HA‐OsPRI2/HA‐OsPRI3/HA‐GUS were immunoprecipitated with GFP‐Trap followed by immunoblotting with the indicated antibodies. HRZ1‐GFP/HA‐GUS, negative control. Protein molecular weight (in kDa) is indicated.

Application: WB    Species: Plant    Sample: Root

FIGURE 1 Interaction of OsHRZ1 with OsPRI2 and OsPRI3. (a) Yeast two‐hybrid assays. Yeast co‐transformed with different BD and AD plasmid combinations were spotted in parallel in 10‐fold dilution series on synthetic dropout medium lacking Leu/Trp/His/Ade. The C‐terminal truncated OsHRZ1 and full‐length OsPRI2/3 were cloned into pGBKT7 and pGADT7, respectively. OsHRZ1‐C/OsPRI1, positive control. OsHRZ1‐ C/Empty, negative control. (b) Pull‐down assay. OsHRZ1 was fused with the GST tag and OsPRI2/3 were fused with the His tag. Recombinant proteins were expressed in E. coli. Proteins were pulled down by glutathione Sepharose 4B and detected using the anti‐His antibody. Protein molecular weight (in kDa) is indicated. (c) CoIP assay. Total proteins from different combinations with OsHRZ1‐GFP and HA‐OsPRI2/HA‐OsPRI3/ HA‐GUS were immunoprecipitated with GFP‐Trap followed by immunoblotting with the indicated antibodies. HRZ1‐GFP/HA‐GUS, negative control. Protein molecular weight (in kDa) is indicated. (d) Degradation of OsPRI2 or OsPRI3 was carried out by detecting the OsPRI2/3‐GFP protein level in co‐infiltration experiments with increasing amounts of OsHRZ1‐GFP. GFP proteins were used as an internal control. Anti‐GFP antibody was used in western blot. Protein molecular weight (in kDa) is indicated. Stars indicate the non‐specific bands. Numbers indicate the ratio of the concentrations of agrobacteria used in co‐infiltration. Empty vector, a binary vector pOCA30 with a 35S promoter; GFP, 35S:GFP in pOCA30; OsPRI2‐GFP, 35S:OsPRI2‐GFP in pOCA30; OsPRI3‐GFP, 35S:OsPRI3‐GFP in pOCA30; OsHRZ1‐GFP, 35S:OsHRZ1‐GFP in pOCA30. (e) Cell‐free degradation. Ten‐day‐old roots grown in Fe‐sufficient solution were harvested and used for protein extraction. Incubation with or without MG132 was performed over the indicated time course

3). Yin X et al. The Nuclear-Localized RxLR Effector PvAvh74 From Plasmopara viticola Induces Cell Death and Immunity Responses in Nicotiana benthamiana. Front Microbiol 2019 Jul 10;10:1531 (PubMed: 31354650) [IF=4.235]

4). Yin X et al. The Nuclear-Localized RxLR Effector PvAvh74 From Plasmopara viticola Induces Cell Death and Immunity Responses in Nicotiana benthamiana. Front Microbiol 2019 Jul 10;10:1531 (PubMed: 31354650) [IF=4.235]

5). Duan Z et al. Chicken bromodomain-containing protein 2 interacts with the Newcastle disease virus matrix protein and promotes viral replication. Vet Res 2020 Sep 22;51(1):120. (PubMed: 32962745) [IF=3.357]

Application: WB    Species: Chicken    Sample: DF-1 cells

Figure 1| Identifcation of the interaction between the NDV M protein and chBRD2 protein.B Identifcation of the interaction between the M protein and chBRD2 protein by co-IP assay. The indicated plasmids were co-transfected into DF-1 cells, and reciprocal co-IP assays were performed to identify the interaction between HA-M and Myc-chBRD2 in DF-1 cells at 36 hpt.

6). Li C et al. Porcine TRIM35 positively regulate TRAF3-mediated IFN-β production and inhibit Japanese encephalitis virus replication. Dev Comp Immunol 2022 Feb;127:104290. (PubMed: 34626690) [IF=3.192]

7). Sun L et al. Proteasome inhibition promotes mono-ubiquitination and nuclear translocation of mature (52 kDa) PINK1. Biochem Biophys Res Commun 2019 Sep 17;517(2):376-382. (PubMed: 31362890)

8). Chen F et al. Assessment of SENP3-interacting proteins in hepatocytes treated with diethylnitrosamine by BioID assay. Acta Biochim Biophys Sin (Shanghai) 2021 Aug 31;53(9):1237-1246. (PubMed: 34312671)

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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.

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