Ab2237
Manufactured by Abcam
Sourced in United Kingdom, United States
Ab2237 is a primary antibody used for immunodetection. It is a polyclonal antibody raised against a specific target antigen. The antibody can be used in various immunoassay techniques, such as Western blotting, ELISA, and immunohistochemistry, to detect and quantify the target protein.
Automatically generated - may contain errors
Lab products found in correlation
Ab32053, by Abcam (2 mentions)
Ab205718, by Abcam (2 mentions)
Ab81552, by Abcam (2 mentions)
Ab8895, by Abcam (1 mentions)
Ab8580, by Merck Group (1 mentions)
Ab817, by Abcam (1 mentions)
Bs1454, by Bioworld Technology (1 mentions)
Scn400 slide scanner, by Leica Microsystems (1 mentions)
Inverted fluorescence microscope, by Leica camera (1 mentions)
Dab staining system, by Beyotime (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Beyotime (1 mentions)
Ab150077, by Abcam (1 mentions)
Ab15497, by Abcam (1 mentions)
Bca protein assay kit, by Beyotime (1 mentions)
Ab133273, by Abcam (1 mentions)
Chemiluminescence detection system, by GE Healthcare (1 mentions)
Ab8226, by Abcam (1 mentions)
Ab198394, by Abcam (1 mentions)
Goat anti rabbit igg h l hrp, by Abcam (1 mentions)
Ripa lysis and extraction buffer, by Thermo Fisher Scientific (1 mentions)
7 protocols using ab2237
1
Chromatin Immunoprecipitation Protocol
2
Immunohistochemical analysis of SIAH1, YBX-1, and Ki-67
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Immunohistochemistry (IHC) staining were conducted as previously described using Goat monoclonal anti- SIAH1 (Abcam, #ab2237, dilutions: 1:250), Rabbit monoclonal anti- YBX-1 (Abcam, #ab12148, dilutions: 1:1000) or Rabbit anti-Ki-67 (Bioworld, #bs1454, dilutions: 1:200). Images were taken using SCN400 Slide Scanner (Leica Microsystems).
3
Immunohistochemical and Immunofluorescence Analysis
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Paraffin-embedded slides of villous tissue were dewaxed in xylol for 20 minutes and rinsed with 100% ethanol for the staining process. Each slide was incubated with Ki-67 antibody (1:100, #2642-1, Abcam, RRID: AB_1580751 ) for 24 hours at 4 °C. Immunostaining was visualized with the DAB staining system (Beyotime). HTR-8/SVneo cells were fixed and permeabilized with 4% paraformaldehyde containing 0.5% Triton X-100 for 20 minutes. Fixed cells were washed and blocked in 1% BSA for 1 hour and incubated with anti-SIAH1 (1:200, #ab2237, Abcam, RRID: AB_2270373 ) and anti-CCNB1 (1:200, #ab32053, Abcam, RRID: AB_731779 ) antibodies at 4 °C overnight. The cells were washed with PBS and incubated with Alexa Fluor 488 antibody (1:1000; #ab150077, Abcam, RRID: AB_2630356 ). The nuclei were stained with DAPI (Beyotime). Images were visualized using an inverted fluorescence microscope (Leica, Wetzlar, Germany).
4
Western Blot Analysis of Transfected Cells
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After transfection, HTR-8/SVneo cells or JEG-3 cells were treated in lysis buffer. The protein concentration was quantified by a BCA Protein Assay Kit (Beyotime, China). Subsequently, the proteins were transferred to nitrocellulose membranes, blocked with 2% bovine serum albumin (BSA), and incubated with a specific primary antibody at 4 °C overnight. The membranes were washed and incubated with secondary antibody for 2 hours at room temperature. Finally, the protein bands were visualized using a chemiluminescence detection system. The antibodies against SIAH1 (#ab2237, Abcam, Cambridge, UK, RRID: AB_2270373 ), CCNB1 (ab32053, Abcam, Cambridge, UK, RRID: AB_731779 ), GAPDH (#3683, Cell Signaling Technology, Danvers, MA, USA, RRID:AB_1642205 ), tubulin (#56739, Cell Signaling Technology, Danvers, MA, USA, RRID: AB_2799519 ), proliferating cell nuclear antigen (PCNA) (ab15497, Abcam, Cambridge, UK, RRID: AB_2160360 ), and secondary anti-IgG (ab205718, Abcam, Cambridge, UK, RRID:AB_2819160 ) antibodies were used to determine the protein expression levels. ImageJ (NIH) was used to quantify the proteins.
5
Western Blot Analysis of Prostate Cancer Cells
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22Rv1, VCaP, and LNCaP cells were lysed in RIPA Lysis and Extraction buffer (Thermo Fisher Scientific), and protein concentration was measured using the BCA protein assay kit (Pierce, Rockford, IL, USA). Fifty or 10 μg of protein was loaded onto 10% SDS-PAGE gels, and then transferred to PVDF membranes (Roche, Basel, Switzerland). After blocking in 5% skimmed milk at room temperature for 1 h, the membranes were incubated with primary antibodies against SIAH1 (1:1,000; Abcam, ab2237), CPSF1 (1:1,000; Abcam, ab81552), AR (1:2,000; Abcam, ab133273), AR-V7 (1:1,000; Abcam, ab198394), and β-actin (1:1,000; Abcam, ab8226) overnight at 4°C, followed by incubation with secondary antibody (goat anti-rabbit IgG H&L [HRP]; 1:1,000; Abcam, ab205718) for 1 h. Immunoblots were visualized using a chemiluminescence detection system (GE Healthcare, Chicago, IL, USA), and luminescent images were scanned into a computer and analyzed using ImageJ software (National Institute of Health, USA).
6
CPSF1 and SIAH1 Protein Interaction
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22Rv1 cells grown in 10-cm dishes at a confluency of 70%–80% were lysed with NP40 buffer. Then, 22Rv1 cell lysates were used for IP with CPSF1 (2 μg/mg; Abcam, ab81552) or SIAH1 (2 μg/mg; Abcam, ab2237) antibody overnight at 4°C. Protein A/G beads (Thermo Fisher Scientific) were added to the IP reactions and left rotating overnight at 4°C. After washing with cold PBS twice, the beads were pelleted by centrifugation at 13,000 rpm at 4°C for 5 min, re-suspended in 10 μL NUPAGE loading buffer, and centrifuged for 2 min at 13,000 rpm before loading onto an SDS gel for western blot analysis.
7
Western Blot Profiling of Wnt Pathway Proteins
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Cells were lysed in an SDS sample buffer and boiled for 5 to 10 min, followed by SDS polyacrylamide gel electrophoresis. Proteins were then transferred to a nitrocellulose membrane and immunoblotted. The primary antibodies used were CK1α (1:1000 ab108296) and SIAH1 (1:250, ab2237) from Abcam, Axin1 (1:1000, 2087), APC (1:1000, 2504), GSK3β (1:1000, 9315), phosphorylated β-Catenin S45 (1:1000, 9564), phosphorylated β-Catenin S33, 37, T41 (1:1000, 9561), β-Catenin (1:1000, 9562), c-Myc (1:5000, 5605), GAPDH (1:5000, 8884), c-Jun (1:5000, 9165), PTEN (1:5000, 9552), and α-Tubulin (1:5000, 9099) from Cell Signaling Technology, α-Tubulin (1:5000, T6199) and GS (1:1000, MAB302) from Millipore, Ubiquitin (1:500, sc-8017) and HSP90 (1:5000, sc-13119) from Santa Cruz Biotechnology, P62 (1:1000, 610832) from BD Biosciences, CRBN (1:5000, NBP1-91810) and MEIS2 (1:250, H00004212-M01) from Novus Biologicals, and Flag (1:5000, F7425) from Sigma (Supplementary Table 2 ). The secondary antibodies used were HRP-conjugated donkey anti-mouse or anti-rabbit (715-035-150 or 711-035-152), or rabbit anti-goat IgG (305-025-045) from Jackson ImmunoResearch (1:10000). ImageJ was used to quantitate the signal of the indicated proteins in immunoblots.
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