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Ab63766

Manufactured by Abcam
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Ab63766 is a recombinant monoclonal antibody specific for Bacterial FtsZ protein. It is produced in Chinese Hamster Ovary (CHO) cells.

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Lab products found in correlation

Odyssey clx system, by LI COR (2 mentions) Ne per nuclear and cytoplasmic extraction kit, by Thermo Fisher Scientific (2 mentions) Savant speedvac system, by Thermo Fisher Scientific (2 mentions) 50kda ultra centrifugation column, by Merck Group (2 mentions) 4 12 bis tris minigels, by Thermo Fisher Scientific (2 mentions) 0.22 pvdf membranes, by Bio-Rad (2 mentions) Mbs854462 anti fgf23, by MyBioSource (2 mentions) Ab8229 anti β actin antibodies, by Abcam (2 mentions) Ab9485, by Abcam (2 mentions) Ab6046, by Abcam (1 mentions) Ab169538, by Abcam (1 mentions) Ab130275, by Abcam (1 mentions) Ab86863, by Abcam (1 mentions) Ab89016, by Abcam (1 mentions) Ab20666, by Abcam (1 mentions) Ab21686, by Abcam (1 mentions) Ab42080, by Abcam (1 mentions) Steponeplus, by Thermo Fisher Scientific (1 mentions) Taqman gene expression assay, by Thermo Fisher Scientific (1 mentions) Ne per nuclear and cytoplasmic extraction reagent, by Thermo Fisher Scientific (1 mentions)

5 protocols using ab63766

1

Antibodies for Immunoblotting, IP, and IF

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Antibodies used for immunoblotting, IP, or immunofluorescence were as follows: anti-TAF7 (TAF7 monoclonal antibody M01, clone 2C5, Abnova); anti-TBP (ab63766, Abcam); anti–β-tubulin (ab6046, Abcam); anti-HA (16B12, ab130275, Abcam); anti-FLAG (clone M2, F3165, MilliporeSigma); anti-puromycin (clone 12D10, MABE343, MilliporeSigma); anti-RPL5 (ab86863, Abcam); anti-RPL8 (ab169538, Abcam); anti-XPO1 (M01180, Boster); anti-eIF5B (ab89016, Abcam); anti-hnRNP U (ab20666, Abcam); anti-Haspin (ab21686, Abcam); anti-SMARCC2 (A301-039A, Bethyl); and anti-DPP9 (ab42080, Abcam). The specificity of the anti-TAF7 antibody for TAF7 has been shown previously using MEFs deleted of TAF7 (13 (link)).
Cheng D., Semmens K., McManus E., Chen Q., Meerzaman D., Wang X., Hafner M., Lewis B.A., Takahashi H., Devaiah B.N., Gegonne A, & Singer D.S. (2021). The nuclear transcription factor, TAF7, is a cytoplasmic regulator of protein synthesis. Science Advances, 7(50), eabi5751.
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2

Protein Extraction and Analysis from Cultured Cells

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Whole femurs were analyzed as previously described.12 (link), 59 (link) Cells were cultured in 75 cm2 flasks for 12 days before treatment. 4 mL culture medium was filtered through a 50KDa ultra centrifugation column (EMD Millipore) and dried overnight in a Savant SpeedVac system (Thermo Fisher Scientific, MA, USA). Whole cell lysates were extracted using T-Per lysis buffer containing protease inhibitors cocktail, and nuclear and cytoplasmic extracts using NEPER™ Nuclear and Cytoplasmic Extraction kit (Thermo Fisher Scientific). Whole cell extracts and cytoplasmic lysates were filtered through a 50KDa ultra centrifugation size exclusion column and dried overnight.
Proteins were fractionated on 4-12 Bis-Tris minigels (Thermo Fisher Scientific), transferred on 0.22 PVDF membranes (Bio-Rad Laboratories) and probed with goat polyclonal anti-FGF23 (Immutopics), rabbit polyclonal MBS854462 anti-FGF23 (MyBioSource, CA, USA), rabbit polyclonal ab9485 anti-GAPDH, rabbit polyclonal ab2185 anti-HIF1α, rabbit polyclonal ab63766 anti-TATA binding protein TBP, and goat polyclonal ab8229 anti- β-actin antibodies (abcam). Antigen antibody complexes were visualized by immunoblotting using anti-goat and anti-rabbit IR fluorescent antibodies on an ODYSSEY® CLx system (LI-COR, NE, USA).
David V., Martin A., Isakova T., Spaulding C., Qi L., Ramirez V., Zumbrennen-Bullough K.B., Sun C.C., Lin H.Y., Babitt J.L, & Wolf M. (2016). Inflammation and functional iron deficiency regulate fibroblast growth factor 23 production. Kidney international, 89(1), 135-146.
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3

Protein Extraction and Analysis from Cultured Cells

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Whole femurs were analyzed as previously described.12 (link), 59 (link) Cells were cultured in 75 cm2 flasks for 12 days before treatment. 4 mL culture medium was filtered through a 50KDa ultra centrifugation column (EMD Millipore) and dried overnight in a Savant SpeedVac system (Thermo Fisher Scientific, MA, USA). Whole cell lysates were extracted using T-Per lysis buffer containing protease inhibitors cocktail, and nuclear and cytoplasmic extracts using NEPER™ Nuclear and Cytoplasmic Extraction kit (Thermo Fisher Scientific). Whole cell extracts and cytoplasmic lysates were filtered through a 50KDa ultra centrifugation size exclusion column and dried overnight.
Proteins were fractionated on 4-12 Bis-Tris minigels (Thermo Fisher Scientific), transferred on 0.22 PVDF membranes (Bio-Rad Laboratories) and probed with goat polyclonal anti-FGF23 (Immutopics), rabbit polyclonal MBS854462 anti-FGF23 (MyBioSource, CA, USA), rabbit polyclonal ab9485 anti-GAPDH, rabbit polyclonal ab2185 anti-HIF1α, rabbit polyclonal ab63766 anti-TATA binding protein TBP, and goat polyclonal ab8229 anti- β-actin antibodies (abcam). Antigen antibody complexes were visualized by immunoblotting using anti-goat and anti-rabbit IR fluorescent antibodies on an ODYSSEY® CLx system (LI-COR, NE, USA).
David V., Martin A., Isakova T., Spaulding C., Qi L., Ramirez V., Zumbrennen-Bullough K.B., Sun C.C., Lin H.Y., Babitt J.L, & Wolf M. (2016). Inflammation and functional iron deficiency regulate fibroblast growth factor 23 production. Kidney international, 89(1), 135-146.
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4

Comprehensive RNA and Protein Analysis Pipeline

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Cell line and tumor total RNA was extracted using RNAzol (Sigma) according to the manufacturer’s protocol. RNA (1ug) was used to generate cDNA with the High-Capacity cDNA Reverse Transcription Kit (Thermo). For qPCR, we used the StepOnePlus instrument (Thermo) with pre-designed TaqMan gene expression assays (Thermo): CXCR4 (Hs00607978_s1), DARS (Hs00154683_m1), IL6 (Hs00174131_m1), IL8 (Hs00174103_m1) and TBP (Hs00427620_m1). TBP served as housekeeping control and data was analyzed using the double delta Ct method. For immunoblotting sub-confluent cells were pelleted, washed with cold PBS and lysed with RIPA buffer. Nuclear and cytoplasmic extracts were obtained using the NE-PER Nuclear and Cytoplasmic extraction reagents (Thermo) according to the recommendations of the manufacturer. Proteins were separated by SDS-PAGE, transferred to PVDF membranes and blotted with antibodies against CXCR4 (Abcam, ab124824, 1:500), HIF2α (Novus, NB100-122, 1:500), VHL (Cell Signaling, 2738, 1:500), phospho-p65 (Cell Signaling, 3033, 1:1000), p65 (Santa Cruz, sc8008, 1:1000), TBP (Abcam, ab63766, 1:5000), B-tubulin (Abcam, ab6046, 1:10000) and ß-actin (Sigma, A5441, 1:5000), ß-tubulin (Sigma, T5201, 1:5000). Secondary antibodies were HRP-conjugated (Dako, 1:5000).
Rodrigues P., Patel S.A., Harewood L., Olan I., Vojtasova E., Syafruddin S.E., Zaini M.N., Richardson E.K., Burge J., Warren A.Y., Stewart G.D., Saeb-Parsy K., Samarajiwa S.A, & Vanharanta S. (2018). NF-kappaB-dependent lymphoid enhancer co-option promotes renal carcinoma metastasis. Cancer discovery, 8(7), 850-865.
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5

Western Blot Analysis of Caco-2BBE Cell Proteins

Cited 2 times
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Caco-2BBE cells were washed with sterile phosphate-buffered saline and treated with RIPA buffer containing proteinase inhibitors (Sigma-Aldrich, St. Louis, MO, USA). Cell lysates were prepared and subjected to western blotting as previously described (Moon et al. 2022 (link); Um et al. 2023 (link)) using antibodies against ChREBP (NB400-13; Novus Biologicals, Centennial, CO, USA), GLUT5 (27571-1-AP; Proteintech, Rosemont, IL, USA), GAPDH (MAB374; Millipore, St. Louis, MO, USA), and TATA-binding protein (TBP) (ab63766; Abcam, Cambridge, UK). To detect nuclear ChREBP, nuclear lysates of Caco-2BBE cells were obtained using the NE-PER™ Nuclear and Cytoplasmic Extraction Reagents Kit (78833; Thermo Fisher Scientific, Carlsbad, CA, USA), according to the manufacturer’s instructions. Immunoreactive proteins were detected using an ECL kit (Thermo Fisher Scientific, Carlsbad, CA, USA), and the images were processed using Adobe Photoshop (Adobe, San Jose, CA, USA). Immunoreactive proteins were visualized using Amersham ImageQuant 800 (Marlborough, MA, USA). Protein bands were quantified using ImageJ software (National Institutes of Health, Bethesda, MD, USA).
Hwang S., Park S., Kim J., Oh A.R., Lee H.J, & Cha J.Y. (2024). Role of Carbohydrate response element-binding protein in mediating dexamethasone-induced glucose transporter 5 expression in Caco-2BBE cells and during the developmental phase in mice. Animal Cells and Systems, 28(1), 15-24.
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