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Ab169639

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Ab169639 is a primary antibody that recognizes the SWI/SNF-related matrix-associated actin-dependent regulator of chromatin subfamily A member 5 (SMARCA5) protein. SMARCA5 is a DNA-dependent ATPase that is a component of the ISWI chromatin remodeling complex. This antibody can be used for applications such as Western blotting and immunohistochemistry.

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Lab products found in correlation

Anti mouse alexa 488, by Thermo Fisher Scientific (1 mentions) Vectashield, by Vector Laboratories (1 mentions) Rpn2106, by Cytiva (1 mentions) Odyssey v1, by LI COR (1 mentions) Iblot, by Thermo Fisher Scientific (1 mentions) A5441, by Merck Group (1 mentions) Horseradish peroxidase, by Merck Group (1 mentions) Ab124741, by Abcam (1 mentions) Ab81292, by Abcam (1 mentions) Ab18192, by Abcam (1 mentions) Alexa fluor 488, by Thermo Fisher Scientific (1 mentions) T6199, by Merck Group (1 mentions) Alexa fluor 555, by Thermo Fisher Scientific (1 mentions)

3 protocols using ab169639

1

Immunostaining of Cellular Structures

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Preparation and immunostaining of cells was performed using standard protocol as described [34 (link)]. The primary antibody used was anti-Coilin (1:2000; Ff-7, mouse sc-55594, Santa Cruz, USA)), anti-PARG (1:1000; rabbit, ab169639, Abcam, USA), and the secondary antibody used was goat anti-rabbit Alexa-563 (1:400, Molecular Probes, USA) and anti-mouse Alexa-488 (1:400, Molecular Probes, USA). Slides were mounted in Vectashield (Vector Laboratories, Burlingame, CA, USA) with Draq5 dye (424101, BioLegend, USA) at 0.05 mg/ml for DNA staining.
Karpova Y., Wu C., Divan A., McDonnell M.E., Hewlett E., Makhov P., Gordon J., Ye M., Reitz A., Childers W.E., Skorski T., Kolenko V, & Tulin A.V. (2019). Non-NAD-like PARP-1 inhibitors in prostate cancer treatment. Biochemical pharmacology, 167, 149-162.
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2

Quantitative Protein Analysis by Western Blot

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For semi-quantitative protein analysis, cells were lysed in 1xSDS sample buffer (25 mM Tris (pH 6.8), 2% 2-mercaptoethanol, 3% SDS, 0.1% bromophenol blue, and 5% glycerol) at 1 × 107 cells/ml and then boiled for 5 min. Proteins were resolved by SDS-PAGE and transferred to i-Blot (IB23001, Invitrogen, USA). The following antibodies were used for immunoblotting assays: anti-pADPr (1:4000, rabbit, 528815, Calbiochem, USA), anti-pADPr (1:500, mouse, 1020, Tulip Biolabs, USA), anti-hPARG (1:1000; rabbit, ab169639 Abcam, USA), anti PSA (1:1000, rabbit, 5153, Cell Signaling Technology, USA), anti-β-actin (1:5000, mouse, A5441, Sigma, USA), anti-GFP (1:1000, rabbit, TP401, Origene, USA), anti-GFP (1:5000, mouse, 632380, BD, USA), and either goat anti-rabbit or anti-mouse secondary antibody conjugated to horseradish peroxidase (Sigma, USA). Western blotting was done using the detection kit from Amersham/GE Healthcare (RPN2106, USA), according to the manufacturer’s instructions. Image digitizing and quantitative analysis were performed by Odyssey v1.2 software (LI-COR, Lincoln, NE, USA).
Karpova Y., Wu C., Divan A., McDonnell M.E., Hewlett E., Makhov P., Gordon J., Ye M., Reitz A., Childers W.E., Skorski T., Kolenko V, & Tulin A.V. (2019). Non-NAD-like PARP-1 inhibitors in prostate cancer treatment. Biochemical pharmacology, 167, 149-162.
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3

DNA Damage Response Pathway Knockdown

Cited 1 time
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The Human ON-TARGETplus siRNA Library-DNA Damage Response and the siRNA targeting OGG1, PARG, Pol β, and XRCC1, each containing a pool of four different siRNA sequences to help aid knockdown efficiencies, were from Horizon Discovery (Cambridge, UK). The non-targeting control siRNA (AllStars Negative Control siRNA) was from Qiagen (Manchester, UK). The following antibodies were used: γH2AX (05–636; Merck-Millipore, Watford, UK), OGG1, PARG, phosphorylated ATM and phosphorylated DNA-PKcs (ab124741, ab169639, ab81292 and ab18192; Abcam, Cambridge, UK), PARP-1 (sc-53643; Santa Cruz Biotechnology, Heidelberg, Germany), OGG1 for immunofluorescence (NB100-106; Bio-Techne Ltd, Abingdon, UK), Polβ and XRCC1 (kindly provided by G. Dianov) and tubulin (T6199; Sigma-Aldrich, Gillingham, UK). Goat anti-mouse Alexa Fluor 555 or goat anti-rabbit Alexa Fluor 488 secondary antibodies for immunofluorescence were from Life Technologies (Paisley, UK). The OGG1 inhibitor TH5487, as previously described [21 (link)], was used at a concentration of 10 µM. The PARG inhibitor PDD00017273, also previously described [22 (link)], was used at a concentration of 1 µM.
Fabbrizi M.R., Nickson C.M., Hughes J.R., Robinson E.A., Vaidya K., Rubbi C.P., Kacperek A., Bryant H.E., Helleday T, & Parsons J.L. (2024). Targeting OGG1 and PARG radiosensitises head and neck cancer cells to high-LET protons through complex DNA damage persistence. Cell Death & Disease, 15(2), 150.
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