701054
antibody from Invitrogen Antibodies
Targeting: MAPT
DDPAC, FLJ31424, FTDP-17, MAPTL, MGC138549, MSTD, MTBT1, MTBT2, PPND, PPP1R103, tau
Antibody data
- Antibody Data
- Antigen structure
- References [2]
- Comments [0]
- Validations
- Western blot [2]
- Immunocytochemistry [1]
- Immunohistochemistry [1]
- Flow cytometry [1]
- Other assay [2]
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- Product number
- 701054 - Provider product page
- Provider
- Invitrogen Antibodies
- Product name
- Phospho-Tau (Ser199) Recombinant Rabbit Monoclonal Antibody (2H23L4)
- Antibody type
- Monoclonal
- Antigen
- Synthetic peptide
- Description
- This antibody is predicted to react with human based on sequence homology.
- Antibody clone number
- 2H23L4
- Concentration
- 0.5 mg/mL
Submitted references Tau Accumulation via Reduced Autophagy Mediates GGGGCC Repeat Expansion-Induced Neurodegeneration in Drosophila Model of ALS.
Differential Hyperphosphorylation of Tau-S199, -T231 and -S396 in Organotypic Brain Slices of Alzheimer Mice. A Model to Study Early Tau Hyperphosphorylation Using Okadaic Acid.
Wen X, An P, Li H, Zhou Z, Sun Y, Wang J, Ma L, Lu B
Neuroscience bulletin 2020 Dec;36(12):1414-1428
Neuroscience bulletin 2020 Dec;36(12):1414-1428
Differential Hyperphosphorylation of Tau-S199, -T231 and -S396 in Organotypic Brain Slices of Alzheimer Mice. A Model to Study Early Tau Hyperphosphorylation Using Okadaic Acid.
Foidl BM, Humpel C
Frontiers in aging neuroscience 2018;10:113
Frontiers in aging neuroscience 2018;10:113
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Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Tau (pS199) was performed by loading 30 µg of mouse brain lysate (lane 1) using Novex®NuPAGE® 4-12% Bis-Tris gel (Product # NP0321BOX), XCell SureLock Electrophoresis System (Product # EI0002), Novex® Sharp Pre-Stained Protein Standard (Product # LC5800), and iBlot® Dry Blotting System (Product # IB21001). Proteins were transferred to a nitrocellulose membrane and blocked with 5% skim milk for 1 hour at room temperature. Tau (pS199) was detected at ~55 kDa using Tau (pS199) Recombinant Rabbit Monoclonal Antibody (Product # 701054) at a 1:1000 dilution in 2.5% skim milk at 4°C overnight on a rocking platform. To confirm specificity, competition was performed with the phosphopeptide (10 µg/mL) (lane 2). Detection was performed using an HRP-conjugated Goat anti-Rabbit secondary antibody (Product # G-21234) at a 1:5000 dilution and chemiluminescent detection was performed using Pierce™ ECL Western blotting Substrate (Product # 32106).
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Western blot analysis of Phospho-Tau pSer199 in total rat brain lysate using a Phospho-Tau pSer199 recombinant rabbit monoclonal antibody (Product # 701054) at a dilution of 1 µg/mL. To confirm specificity, competition was performed by preincubation with the phosphopeptide to inhibit antibody binding (lane 2). Results show a band at ~56kDa.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunofluorescent analysis of Tau (pS199) was done on 70% confluent log phase U-2 OS cells. The cells were fixed with 4% paraformaldehyde for 15 minutes, permeabilized with 0.25% Triton X-100 for 10 minutes, and blocked with 5% BSA for 1 hour at room temperature. The cells were labeled with Tau (pS199) Recombinant Rabbit Monoclonal Antibody (Product # 701054) at a dilution of 1:500 in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Fluor® 488 Goat anti-Rabbit IgG Secondary Antibody (Product # A-11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 Phalloidin (Product # A12381) and Panel d is a merged image showing nuclear and cytoplasmic localization and panel e shows competition with the phospho-Tau (pS199) peptide. The images were captured using a Nikon microscope at 20X magnification.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Immunohistochemistry analysis of Phospho-Tau pSer199 showing staining in the cytoplasm of paraffin-embedded rat cerebellum tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10 mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with Phospho-Tau pSer199 Monoclonal antibody (Product # 701054) diluted in 3% BSA-PBS at a dilution of 1:200 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using a HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Flow cytometry analysis of Tau (pS199) was performed on HeLa cells. Cells were fixed with 70% ethanol for 10 minutes, permeabilized with 0. 25% Tritonª X-100 for 20 minutes, and blocked with 5% BSA for 1 hour at room temperature. Cells were labeled with ABfinityª Tau (pS199) recombinant rabbit monoclonal antibody (Product # 701054, red histogram) or with rabbit isotype control (pink histogram) at a dilution of 1:250 in 2.5% BSA. After incubation at room temperature for 3 hours, the cells were labeled with Alexa Fluor¨ 488 goat anti-Rabbit Secondary antibody (Product # A11008) at a dilution of 1:400 for 30 minutes at room temperature. The representative 10,000 cells were acquired and analyzed for each sample using an Attune¨ Acoustic Focusing Cytometer. The purple histogram represents unstained control cells and the green histogram represents no-primary-antibody control.
Supportive validation
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 2 Tau phosphorylation in vitro . One microgram recombinant human tau (left, A) or mouse tau (right, A) was incubated with glycogensynthase-kinase-3beta (GSK-3ss) and ATP in kinase buffer and analyzed by Western blot using antibodies against total tau (tau-5) phospho-tau-S199, phospho-tau-T231 and phospho-tau-S396. Lanes B and C served as a control omitting either the enzyme GSK-3beta (Lane B) or tau protein (Lane C). Size markers are given as kDa.
- Submitted by
- Invitrogen Antibodies (provider)
- Main image
- Experimental details
- Figure 3 Western blot for p-tau S199. Brain slices of adult WT mice (A) or transgenic (TG) mice (B) were incubated for 14 days at 37degC without (Co) or with Okadaic acid (OA), wortmannin (WM), or combination of OA+WM. Extracted brain slices were subsequently analyzed by Western blot using antibodies against total tau (tau-5) and phospho-tau-S199. Size markers are given as kDa on the right side. Actin served as a control.