Product Info

Source:
Mouse
Application:
WB 1:3000-1:10000, IHC 1:50-1:200, IF/ICC: 1:100-1:500, IP 1:100-1:500
*The optimal dilutions should be determined by the end user.
Reactivity:
All
Clonality:
Monoclonal [5F67]
Specificity:
GFP-Tag Mouse Monoclonal antibody detects endogenous levels of C-terminal, internal, and N-terminal GFP-tagged proteins.
RRID:
AB_2839413
Cite Format: Affinity Biosciences Cat# T0005, RRID:AB_2839413.
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.

Immunogens

Immunogen:

Full-length GFP protein.

Description:
N/A

References

1). Zhou X et al. Splicing factor SRSF1 promotes gliomagenesis via oncogenic splice-switching of MYO1B. J Clin Invest 2018 Nov 27 (PubMed: 30481162) [IF=19.456]

Application: WB    Species: human    Sample: U87MG and U251 cells

Figure 10.| SRSF1-guided MYO1B splicing determines cell fate through the PDK1/AKT and PAK/LIMK pathways. (D) Co-IP confirmation of the interaction between EGFP-fused MYO1B proteins (MYO1B-fl and MYO1B-t) and endogenous p85 PI3K in U87MG and U251 cells.

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=7.947]

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). Liang L et al. Identification and functional analysis of two new de novo KCNMA1 variants associated with Liang–Wang syndrome. Acta Physiol (Oxf) 2022 Feb 13;e13800. (PubMed: 35156297) [IF=7.523]

4). Zhou Y et al. Orf virus ORF120 protein positively regulates the NF-κB pathway by interacting with G3BP1. J Virol 2021 Jul 21;JVI0015321. (PubMed: 34287041) [IF=6.549]

Application: IF/ICC    Species: Human    Sample: OFTu cells

FIG 1 Generation of the recombinant viruses and their growth properties in vitro. (A) Fluorescence microscopy showing the cytopathic effects (CPEs) of the OFTu cells infected with OV-SY17Δ120 deletion mutant virus by plaque purification. The plaques with a strong fluorescent signal in the OFTu cells infected with OV-SY17Δ120 48 hpi during the second round of plaque purification. (b, e, and h) The same fields as shown in a, d, and g by bright-field microscopy. (c, f, and i) Overlay of the a and b, d and e, or g and h images. The cells were visualized under a microscope at 10× magnification. Bar, 400 μm. (B) A revertant mutant OV-SY17-RV120 was produced using red fluorescent protein (RFP) as a selection marker and obtained after two rounds of plaque assay purification on monolayers of OFTu cell cultures 48 hpi. (b, e, and h) The same fields as in a, d, and g by bright-field microscopy. (c, f, and i) Overlay of the a and b, d and e, or g and h images. The cells were visualized under a microscope at 10× magnification. Bar, 400 μm. (C) PCR was performed to confirm the absence of ORFV ORF120 and the presence of the EGFP reporter gene sequences in the OV-SY17Δ120 genome. The PCR analysis showed that the ORF120 gene was successfully deleted in the OV-SY17Δ120 mutant. M, molecular marker; lanes 1 to 3, three OV-SY17Δ120 recombinants from plaque purification; lane 4, wild-type OV-SY17. (D) PCR was performed to confirm the presence of both the ORF120 and RFP reporter genes in the OV-SY17-RV120 genome. The PCR analysis showed that the corresponding revertant virus OV-SY17-RV120 was successfully rescued. M, molecular marker; lanes 1 to 3, three OV-SY17-RV120 recombinants from plaque purification; lane 4, wild-type OV-SY17. (E) In vitro growth kinetics of the OV-SY17Δ120 mutant virus, OV-SY17 parental virus, and OV-SY17-RV120 revertant virus. Primary OFTu cells were infected (MOI, 10 or 0.1) with either the OV-SY17Δ120 mutant virus, parental virus OV-SY17, or revertant virus OV-SY17-RV120 and the virus titers were determined by TCID50 assays at the indicated postinfection time points. The growth curves of the OV-SY17Δ120 mutant virus were similar to those of the OV-SY17 parental virus and the OV-SY17-RV120 revertant virus, indicating that the deletion of ORF120 did not affect virus replication.

Application: WB    Species: Human    Sample: OFTu cells

FIG 1 Generation of the recombinant viruses and their growth properties in vitro. (A) Fluorescence microscopy showing the cytopathic effects (CPEs) of the OFTu cells infected with OV-SY17Δ120 deletion mutant virus by plaque purification. The plaques with a strong fluorescent signal in the OFTu cells infected with OV-SY17Δ120 48 hpi during the second round of plaque purification. (b, e, and h) The same fields as shown in a, d, and g by bright-field microscopy. (c, f, and i) Overlay of the a and b, d and e, or g and h images. The cells were visualized under a microscope at 10× magnification. Bar, 400 μm. (B) A revertant mutant OV-SY17-RV120 was produced using red fluorescent protein (RFP) as a selection marker and obtained after two rounds of plaque assay purification on monolayers of OFTu cell cultures 48 hpi. (b, e, and h) The same fields as in a, d, and g by bright-field microscopy. (c, f, and i) Overlay of the a and b, d and e, or g and h images. The cells were visualized under a microscope at 10× magnification. Bar, 400 μm. (C) PCR was performed to confirm the absence of ORFV ORF120 and the presence of the EGFP reporter gene sequences in the OV-SY17Δ120 genome. The PCR analysis showed that the ORF120 gene was successfully deleted in the OV-SY17Δ120 mutant. M, molecular marker; lanes 1 to 3, three OV-SY17Δ120 recombinants from plaque purification; lane 4, wild-type OV-SY17. (D) PCR was performed to confirm the presence of both the ORF120 and RFP reporter genes in the OV-SY17-RV120 genome. The PCR analysis showed that the corresponding revertant virus OV-SY17-RV120 was successfully rescued. M, molecular marker; lanes 1 to 3, three OV-SY17-RV120 recombinants from plaque purification; lane 4, wild-type OV-SY17. (E) In vitro growth kinetics of the OV-SY17Δ120 mutant virus, OV-SY17 parental virus, and OV-SY17-RV120 revertant virus. Primary OFTu cells were infected (MOI, 10 or 0.1) with either the OV-SY17Δ120 mutant virus, parental virus OV-SY17, or revertant virus OV-SY17-RV120 and the virus titers were determined by TCID50 assays at the indicated postinfection time points. The growth curves of the OV-SY17Δ120 mutant virus were similar to those of the OV-SY17 parental virus and the OV-SY17-RV120 revertant virus, indicating that the deletion of ORF120 did not affect virus replication.

5). 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]

6). Xu Z et al. Mutated SASH1 promotes Mitf expression in a heterozygous mutated SASH1 knock‑in mouse model. Int J Mol Med 2020 Jun 19. (PubMed: 32582980) [IF=5.314]

7). 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=3.829]

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

Figure 5 |siRNA-mediated knockdown of chBRD2 promotes NDV replication in DF-1 cells.F The expression levels of the M and GFP proteins in chBRD2 siRNA#2- or control siRNA-treated cells were examined by western blot at 6, 12 and 24 hpi, respectively. The relative expression levels of the M and GFP proteins compared to control GAPDH expression levels were determined by densitometry using ImageJ software version 1.8.0. Error bars represent standard deviations(mean±SD) (*P<0.05; **P<0.01 compared to the value of the control siRNA group).

8). 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) [IF=3.322]

9). Zhuang T et al. Eukaryotic expression and immunogenicity of Ancylostoma ceylanicum calreticulin. Parasitol Int 2022 Jul 21;91:102633. (PubMed: 35872299)

10). Ben-Chang Li et al. Plasmalemma localisation of DOUBLE HYBRID PROLINE-RICH PROTEIN 1 and its function in systemic acquired resistance of Arabidopsis thaliana. Functional Plant Biology 2014;41(7):768.

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