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P stat3 tyr705 antibody

Manufactured by Cell Signaling Technology
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The P-STAT3 (Tyr705) antibody is a tool used to detect and quantify the phosphorylation of STAT3 at tyrosine 705. STAT3 is a transcription factor that plays a key role in cellular signaling pathways. Phosphorylation of STAT3 at tyrosine 705 is a crucial step in its activation and subsequent regulation of target genes.

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Lab products found in correlation

Bca kit, by Beyotime (1 mentions) Stat3 antibody, by Cell Signaling Technology (1 mentions) Gapdh, by Cell Signaling Technology (1 mentions) Dako envision dual link system hrp dab, by Agilent Technologies (1 mentions) Eclipse ti microscope, by Nikon (1 mentions) Tele pack vet x led endoscopic video unit, by Storz (1 mentions) Bond rx autostainer, by Leica (1 mentions) Vectastain universal elite abc kit, by Vector Laboratories (1 mentions)

Spelling variants of this product

STAT3 (1075 citations) P-STAT3 (935 citations) P-STAT3 (Tyr705) (157 citations) STAT3 antibody (37 citations) P-STAT3 antibody (16 citations) Anti-p-STAT3 antibody (Tyr705) (3 citations)

5 protocols using p stat3 tyr705 antibody

1

Protein Extraction and Western Blotting

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A total protein extraction kit (Beyotime Bio-technology) was used to extract total protein from different groups, and protein concentration was determined using the BCA kit (P0012, Beyotime Bio-technology, Shanghai, China). From each group, a 20 μg protein sample was electrophoresed on a 10% SDS polyacrylamide gel and then transferred to a polyvinylidene fluoride membrane. After blocking the protein with a protein-free rapid blocking buffer (catalog no. PS108P, EpiZyme, Shanghai, China), polyvinylidene fluoride membranes were incubated for 12 h at 4°C with the following antibodies: USP25 antibody (1: 1,000; catalog no. A7975, ABclonal), STAT3 antibodies (1:1,000; catalog no. #9139, Cell Signaling Technology), p-STAT3 (Tyr705) antibody (1:2,000; catalog no. #9145, Cell Signaling Technology), ZO-1 antibody (1:1,000; catalog no. 21773-1-AP, Proteintech, Wuhan, China), and glyceraldehyde 3-phosphate dehydrogenase (GAPDH) antibody (1:1,000; catalog no. #5174, Cell Signaling Technology).
Liu Z., Qi M., Tian S., Yang Q., Liu J., Wang S., Ji M., Yu R., Zeng S., Li J., Wei Y, & Dong W. (2022). Ubiquitin-Specific Protease 25 Aggravates Acute Pancreatitis and Acute Pancreatitis-Related Multiple Organ Injury by Destroying Tight Junctions Through Activation of The STAT3 Pathway. Frontiers in Cell and Developmental Biology, 9, 806850.
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2

Kidney Tissue Immunohistochemistry Protocol

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Kidney slices were fixed in alcohol-formalin-acetic acid, dehydrated in ethanol and xylene, embedded in paraffin, and cut into 3 mm sections. Samples were then deparaffinized, rehydrated, and heated at 97°C for 20 minutes in citrate buffer. Endogenous peroxidase was inactivated by incubation in 0.3% H2O2 for 10 minutes at room temperature. Sections were incubated with PBS containing anti-HSPA5 (BiP, GRP78, sc-1050, Santa Cruz Biotechnology), anti-ACSL4 (Sigma HPA005552). This antibody has been validated in the Human Protein Atlas project (https://www.proteinatlas.org/ENSG00000068366-ACSL4/summary/antibody). pSTAT3 immunohistochemistry was performed using TBS instead of PBS and p-STAT3 Tyr705 antibody (Cell Signaling CS9145). Subsequently, the sections were incubated with anti-rabbit or anti-goat antibodies conjugated to peroxydase-labeled polymer and visualized with a peroxydase kit (Dako EnVision®+ Dual Link System-HRP (DAB+), Agilent). Finally, tissue sections were counterstained with hematoxylin. The samples were visualized using a Nikon Eclipse Ti microscope.
Poindessous V., Lazareth H., Crambert G., Cheval L., Sampaio J.L, & Pallet N. (2024). STAT3 drives the expression of ACSL4 in acute kidney injury. iScience, 27(6), 109737.
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3

Identifying IL-21 Responsive Cells in Kidney

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For identification of IL-21 responsive cell types in kidneys, freshly prepared kidney single cell suspensions were incubated with rIL-21 (20 ng/ml, R&D systems) for 30 min or left untreated. Cells were fixed with 1% paraformaldehyde, permeabilized with 90% methanol and washed. Aliquots of untreated cells were dephosphorylated with λ protein phosphatase (NEB BioLabs). All samples were then stained with p-STAT3 (Tyr705) antibody (Cell Signaling). To determine the endogenous phosphorylation state of Akt and S6 in renal T cell populations, kidneys were directly disrupted in RPMI containing 1% paraformaldehyde. Fixed cells were permeabilized with 90% methanol, washed and aliquots of all samples were incubated with λ protein phosphatase. Cells were stained with p-Akt (Ser308), p-Akt (Thr473) or p-S6 (Ser235/236) antibody (Cell Signaling).
Teichmann L.L., Cullen J.L., Kashgarian M., Dong C., Craft J, & Shlomchik M.J. (2015). Local triggering of the ICOS coreceptor by CD11c+ myeloid cells drives organ inflammation in lupus. Immunity, 42(3), 552-565.
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4

Immunohistochemical Analysis of pSTAT3 Activation

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Immunohistochemical staining to detect pSTAT3tyr705 was performed on a KPN and an AKPT tumor to determine constitutive activation of the pathway in vivo. A KPN male mouse on a C57BL/6J (generation N5-6) background of 10 weeks of age were induced with a single intraperitoneal injection of 2mg tamoxifen and aged until clinical endpoint as evidenced by anemia, hunching and/or weight loss to generate small intestinal tumor for IHC staining. Intracolonic induction in a 16-week-old female mouse villinCreER Apcfl/+ KrasG12D/+ Trp53fl/fl Tgfbr1fl/fl on a C57BL/6 background (generation N6/N7) was performed under general anesthesia. Three 70µl 100uM dose of 4-hydroxy tamoxifen (H7904-5MG from Sigma) were injected from the mid-colon to distal colon into the sub-mucosa via a colonoscope (Karl Storz TELE PACK VET X LED endoscopic video unit). At clinical endpoint (weight loss with the presentation of hunching) colonic tumor tissue (spontaneous SI tumour) was collected from a single tumour for IHC staining.
Staining was performed using pSTAT3tyr705 antibody (#9131, Cell signaling, Danvers, MA, USA) at 1:100 on a Leica BOND Rx autostainer (Leica Biosystems, Wetzlar, Germany).
Pennel K.A., Hatthakarnkul P., Wood C.S., Lian G.Y., Al-Badran S.S., Quinn J.A., Legrini A., Inthagard J., Alexander P.G., van Wyk H., Kurniawan A., Hashmi U., Gillespie M.A., Mills M., Ammar A., Hay J., Andersen D., Nixon C., Rebus S., Chang D.K., Kelly C., Harkin A., Graham J., Church D., Tomlinson I., Saunders M., Iveson T., Lannagan T.R., Jackstadt R., Maka N., Horgan P.G., Roxburgh C.S., Sansom O.J., McMillan D.C., Steele C.W., Jamieson N.B., Park J.H., Roseweir A.K, & Edwards J. (2024). JAK/STAT3 represents a therapeutic target for colorectal cancer patients with stromal-rich tumors. Journal of Experimental & Clinical Cancer Research : CR, 43, 64.
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5

Immunohistochemical Analysis of p-STAT3 in Bone Marrow

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Immunohistochemistry was performed on paraffin-embedded bone marrow biopsies from 21 patients. Following reduction of endogenous peroxidase activity, deparaffinized slides were incubated at 4°C overnight with a rabbit anti-human phosphorylated STAT3 (p-STAT3 Tyr705) antibody diluted at 1:50 (clone D3A7; Cell Signaling Technology). Labeling was revealed by the R.T.U. VectaStain universal Elite ABC kit (Vector Laboratories). Briefly, after two washes, slides were incubated with a horse serum blocking solution included in the kit for 20 min. After washing, slides were incubated with a secondary biotinylated antibody, then stained with diaminobenzidine substrate (Novocastra) during 3 min. Slides were then washed twice with distilled water, counterstained with 1% hematoxylin and analyzed under a light microscope at a 40x magnification by two independent investigators in a blinded fashion. Positively stained plasma cells were evaluated for each sample.
Beldi-Ferchiou A., Skouri N., Ben Ali C., Safra I., Abdelkefi A., Ladeb S., Mrad K., Ben Othman T, & Ben Ahmed M. (2017). Abnormal repression of SHP-1, SHP-2 and SOCS-1 transcription sustains the activation of the JAK/STAT3 pathway and the progression of the disease in multiple myeloma. PLoS ONE, 12(4), e0174835.
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