Product: HA-Tag Rabbit pAb
Catalog: T0050
Description: Rabbit polyclonal antibody to HA-Tag pAb
Application: WB IP CoIP
Reactivity: All
Uniprot:
RRID: AB_2837583

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Product Info

Source:
Rabbit
Application:
WB 1:3000-1:10000, IP 1:200, CoIP 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:
Polyclonal
Specificity:
The HA tag antibody can recognize C-terminal, internal, and N-terminal HA-tag fusion proteins.
RRID:
AB_2837583
Cite Format: Affinity Biosciences Cat# T0050, RRID:AB_2837583.
Conjugate:
Unconjugated.
Purification:
The antiserum was purified by peptide affinity chromatography using SulfoLink™ Coupling Resin (Thermo Fisher Scientific).
Storage:
1mg/ml in PBS, pH 7.4, containing 0.02% sodium azide and glycerol. Store at -20 °C. Stable for 12 months from date of receipt.

Immunogens

Immunogen:

Rabbit polyclonal antibody is prepared by immunizing synthetic peptide coupled to KLH.

Description:
Human influenza hemagglutinin (HA) is a surface glycoprotein required for the infectivity of the human virus. The HA tag is derived from the HA-molecule corresponding to amino acids 98-106 has been extensively used as a general epitope tag in expression vectors. Many recombinant proteins have been engineered to express the HA tag, which does not appear to interfere with the bioactivity or the biodistribution of the recombinant protein.HA tag antibody is a highly sensitive and affinity monoclonal antibody applicable to HA-tagged fusion protein detection.

References

1). bHLH121 Functions as A Direct Link that Facilitates the Activation of FIT by bHLH IVc Transcription Factors for Maintaining Fe Homeostasis in Arabidopsis. Molecular Plant (PubMed: 31962167) [IF=27.5]

Application: WB    Species: Plant    Sample: tobacco cells

Figure 4. bHLH IVc TFs Promote the Nuclear Accumulation of bHLH121. (A) Subcellular localization of bHLH121. bHLH121-GFP and ER-rk-mCherry (endoplasmic reticulum marker) or NLS-mCherry (nucleus marker) cotransformed into Arabidopsis mesophyll protoplasts. The GFP and mCherry signals were visualized under a confocal microscope. Scale bars, 10 mm. (B) Subcellular localization of bHLH121 in response to Fe deficiency. Plants were grown in +Fe or  Fe medium for 6 days. Roots were stained with propidium iodide (in red). Root maturation regions are shown. Scale bars, 50 mm. (C) Yeast two-hybrid assay. Yeast co-transformed with different BD and AD plasmid combinations was spotted in parallel in a 10-fold dilution series. Growth on selective plates lacking leucine, tryptophan, adenine, and histidine (SD-LWAH) is shown. (D) Co-immunoprecipitation assays. Immunoprecipitation was performed using GFP-Trap agarose beads, and immunoblots were probed with anti-HA antibody (@HA) and anti-GFP antibody (@GFP). (E) bHLH IVc TFs promote the nuclear accumulation of bHLH121. bHLH121-mCherry was co-transformed with GFP, bHLH34-GFP, bHLH104-GFP, bHLH105-GFP, or bHLH115-GFP into tobacco cells. GFP and mCherry signals were visualized under a confocal microscope.

2). PTBP1 knockdown promotes neural differentiation of glioblastoma cells through UNC5B receptor. Theranostics (PubMed: 35664063) [IF=12.4]

3). FER-LIKE FE DEFICIENCY-INDUCED TRANSCRIPTION FACTOR (OsFIT) interacts with OsIRO2 to regulate iron homeostasis. Journal for ImmunoTherapy of Cancer (PubMed: 37295817) [IF=10.9]

4). Oryza sativa POSITIVE REGULATOR OF IRON DEFICIENCY RESPONSE 2 (OsPRI2) and OsPRI3 are involved in the maintenance of Fe homeostasis. PLANT CELL AND ENVIRONMENT (PubMed: 31674679) [IF=7.3]

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

5). Scavenger receptor A impairs interferon response to HBV infection by limiting TRAF 3 ubiquitination through recruiting OTUB 1. The FEBS Journal (PubMed: 31386800) [IF=5.4]

Application: WB    Species: human    Sample: HEK293T cells

Fig. 6.| Scavenger receptor A suppresses TRAF3 K63-linked ubiquitination via recruiting OUTB1.(C) HEK293T cells were transfected with FLAG-TRAF3, together with or without V5-SRA, or with ubiquitin mutants K48 (left panel) and K63 (right panel). The cells were treated with MG132 for 6 h, and then subjected to anti-FLAG antibody pulldown and immunoblotting with anti-HA to evaluate the ubiquitination of TRAF3.

Application: WB    Species: Human    Sample: HepG2.2.15 cells

Fig. 5. Scavenger receptor A colocalizes and interacts with signaling transducer TRAF3. (A) SRA coimmunoprecipitates with TRAF3 in HepG2.2.15 cells. Equal amounts of cell extract were immunoprecipitated with SRA-specific antibody or control IgG. The immune complex was analyzed using anti-TRAF3 antibody (left panel). Besides, TRAF3 was pulled down and analyzed for association with SRA using antiSRA antibody (right panel). Levels of SRA and TRAF3 in the whole cell lysate were also examined. (B) Colocalization of SRA with TRAF3 in HepG2.2.15 cells. The cells were cultured on glass coverslips and immunostained with antibodies against SRA and TRAF3, respectively, subsequently analyzed by a confocal microscope (Magnification: 6309). (C) HepG2 cells were transfected with pHBV1.3 or control plasmid, and equal amounts of protein were used for immunoprecipitation with the SRA-specific antibody. The immunoprecipitates were probed for the presence of TRAF3 and SRA, respectively. (D) TRAF-C domain is essential for the binding of TRAF3 to SRA. HEK293T cells were transfected with HA-tagged SRA and FLAG-tagged full-length TRAF3 or several truncated TRAF3 deletion mutants. Immunoprecipitation was performed using anti-FLAG antibody. The presence of coprecipitated SRA was determined by immunoblotting with the anti-HA antibody. Representative results from three independent experiments are shown.

6). TMT-based quantitative proteomics analysis reveals the attenuated replication mechanism of Newcastle disease virus caused by nuclear localization signal mutation in viral matrix protein. Virulence (PubMed: 32420802) [IF=5.2]

7). SRA Suppresses Antiviral Innate Immune Response in Macrophages by Limiting TBK1 K63 Ubiquitination via Deubiquitinase USP15. Microbiology Spectrum (PubMed: 36342281) [IF=3.7]

8). Proteasome inhibition promotes mono-ubiquitination and nuclear translocation of mature (52 kDa) PINK1. BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS (PubMed: 31362890) [IF=3.1]

9). Oryza sativa FER‐LIKE FE DEFICIENCY‐INDUCED TRANSCRIPTION FACTOR (OsFIT/OsbHLH156) interacts with OsIRO2 to regulate iron homeostasis. Journal of Integrative Plant Biology (PubMed: 32237201)

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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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