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HA-Tag antibody Primary Antibody

IF, IP, IHC, ELISA, WB Host: Mouse Monoclonal HA-C5 unconjugated
Pubmed (1 reference) Independent Validation (2 validations)
Catalog No. ABIN2443910
$303.60
Plus shipping costs $45.00
100 μg
local_shipping Shipping to: United States
Delivery in 21 to 26 Business Days
  • Target See all HA-Tag products
    HA-Tag
    Reactivity
    Please inquire
    Host
    • 80
    • 75
    • 14
    • 11
    • 2
    Mouse
    Clonality
    • 92
    • 88
    Monoclonal
    Conjugate
    • 80
    • 17
    • 16
    • 10
    • 5
    • 4
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    • 3
    • 3
    • 3
    • 3
    • 3
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    This HA-Tag antibody is un-conjugated
    Application
    • 130
    • 75
    • 43
    • 29
    • 28
    • 21
    • 15
    • 14
    • 9
    • 7
    • 6
    • 6
    • 4
    • 4
    • 4
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Immunofluorescence (IF), Immunoprecipitation (IP), Immunohistochemistry (IHC), ELISA, Western Blotting (WB)
    Sequence
    YPYDVPDYA
    Purification
    Purified
    Clone
    HA-C5
    Isotype
    IgG3
  • Application Notes
    Optimal working dilution should be determined by the investigator.
    Restrictions
    For Research Use only
  • Validation #100058 (Western Blotting)
    'Independent Validation' Badge
    by
    Georg-August-University of Göttingen, Johann-Friedrich-Blumenbach-Institute for Zoology and Anthropology, Developmental Biology
    No.
    #100058
    Date
    09/16/2016
    Antigen
    HA-Tag
    Lot Number
    16631508212-P
    Method validated
    Western Blotting
    Positive Control
    HA-fusion proteins expressed in NIH/3T3 cells
    Negative Control
    Notes
    Passed, regarding sensitivity and specificity. The HA-tag antibody ABIN2443910 does specifically recognize HA-tagged proteins in whole cell lysates from human tissue culture cells with no obvious background staining.
    'Independent Validation' Badge
    Validation Images
    Full Methods
    Primary Antibody
    ABIN2443910
    Secondary Antibody
    Rabbit anti-Mouse IgG (whole molecule), HRP-linked (Sigma-Aldrich, A9044, lot number: 034M4761)
    Full Protocol
    • Cultured NIH/3T3 cells heterologously expressing either a 26kDa (Fig. A) or 43kDa (Fig. B) HA-tag-fusion protein were rinsed with PBS and lysed in 6M Urea/20mM Tris.
    • 40µg total protein were used for Western blot analysis.
    • Proteins were denatured and separated on 10% SDS-PAGE (Laemmli 1970) and blotted to Amersham Protran Premium 0.2 NC (GE Healthcare, 10600004, lot A10043108) (Towbin et al., 1979).
    • Blocking of membrane in 5% skim milk in TBST (50 mM Tris-HCl, pH 7.4, 150mM NaCl, 0.1% Tween 20) for 30min at RT.
    • Incubation with primary antibody ABIN2443910 diluted 1:600 in 5% skim milk in TBST overnight at 4°C.
    • Washing in TBST for 30min at RT.
    • Incubation with secondary antibody secondary antibody: rabbit-anti-mouse IgG (whole molecule), HRP-linked (Sigma-Aldrich, A9044, lot 034M4761) diluted 1:10000 in 5% skim milk in TBST for 45min at RT.
    • Washing in TBST for 30-45min at RT.
    • Chemiluminescence detection using Clarity Western ECL Substrate (BioRad, 170-5061) according to the supplier’s recommendations and image capture via X-ray films.
    Experimental Notes
    A dilution factor <1000 is recommended for Western blot on cellular lysates with ABIN2443910.
  • Validation #100059 (Immunocytochemistry)
    'Independent Validation' Badge
    by
    Georg-August-University of Göttingen, Johann-Friedrich-Blumenbach-Institute for Zoology and Anthropology, Developmental Biology
    No.
    #100059
    Date
    09/16/2016
    Antigen
    HA-Tag
    Lot Number
    16631508212-P
    Method validated
    Immunocytochemistry
    Positive Control
    HA-fusion proteins transiently expressed in NIH/3T3 cells
    Negative Control
    Notes
    Passed, regarding sensitivity and specificity. The antibody specifically detects the HA-fusion protein in immuno-cytochemical experiments. No obvious background staining was found.
    'Independent Validation' Badge
    Validation Images
    Full Methods
    Primary Antibody
    ABIN2443910
    Secondary Antibody
    MFP488 goat anti-mouse IgG (H+L) (MoBiTec, MFP-A1029, lot 2803101)
    Full Protocol
    • NIH/3T3 cells (ATCC) were grown on cover slips in DMEM, 10% fetal bovine serum, 5% penicillin/streptomycin (all Gibco) at 37°C in 5% CO2.
    • Cells were transfected using Metafectene Pro (Biontex, T040-1.0, lot AD1.13) according to the manual with a plasmid encoding an HA-tag fusion protein that is expected to localize to the nuclear membrane and the cytoplasm.
    • 24h post-transfection cells were fixed in 3.7% paraformaldehyde in PBS for 15min and processed for immuno-cytology.
    • Unspecific binding sites were blocked in PBST (phosphate buffered saline containing 0.15% bovine serum albumin, 0.1% Tween-20) for 1h.
    • Cells were incubated with the primary antibody ABIN2443910 diluted 1:100 in PBS at 4°C overnight.
    • Cells were washed in TBST (50mM Tris-HCl, pH 7.4, 150mM NaCl, 0.1% Tween 20) for 15min.
    • Incubation with the secondary antibody MFP488 goat anti-mouse IgG (H+L) (MoBiTec, MFP-A1029, lot 2803101) diluted 1:600 in PBS and DAPI (stock solution 10µg/µl, diluted 1:500) for 1h at 37°C.
    • Cells were washed in TBST for 15min.
    • Cells were embedded in Fluorescent Mounting Medium (Dako, S3023, lot 10090890).
    • Images were taken by confocal microscopy (LSM 510, Zeiss), and processed using Adobe Photoshop 5.0.
    Experimental Notes
    none
  • Storage
    4 °C/-20 °C/-80 °C
    Storage Comment
    Lyophilized antibodies can be kept at 4ºC for up to 3 months and should be kept at -20ºC for long-term storage (2 years). To avoid freeze-thaw cycles, reconstituted antibodies should be aliquoted before freezing for long-term (1 year) storage (-80ºC) or kept at 4ºC for short-term usage (2 months).
  • Milewska, Kindler, Vkovski, Zeglen, Ochman, Thiel, Rajfur, Pyrc: "APOBEC3-mediated restriction of RNA virus replication." in: Scientific reports, Vol. 8, Issue 1, pp. 5960, (2019) (PubMed).

  • Target
    HA-Tag
    Alternative Name
    HA Tag (HA-Tag Products)
    Target Type
    Tag
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