Whole-cell protein extracts were obtained by direct lysis in SDS sample buffer heated to 75°C. Proteins resolved by SDS-PAGE were transferred to a polyacrylamide membrane (Bio-rad laboratories, Calfornia, USA.) and probed with antibodies raised against 11βHSD1, 1:5000 (AB83552, Abcam, UK); CRABP2, 1:5000 (HPA004135, SIGMA, Stockholm, Sweden); FABP5, 1:1000 (AB37267, Abcam, UK); RARα, 1:1700, (AB28767, Abcam, UK); RXRα,1:1000 (5388S, Cell Signaling, Danvers, USA); PPARβ/δ, 1:1000 (AB8937, Abcam, UK); Cyp26a1, 1:2000 (SC53618, SantaCruz); DHRS3, 1:200 (15393-1-AP; Proteintech Group Manchester, UK); RBP4,1:1000 (SC46688, SantaCruz); RDH12, 1:1000 (AB87038, Abcam, UK); and β-actin,1:5000 (A2066, SIGMA, UK). After incubation with peroxidase-conjugated secondary antibody, anti-rabbit or anti-mouse IgG diluted 1:1000–1:50,000 (Jackson Immunoresearch Laboratories, West Grobe, USA), detection of chemiluminescence reaction was visualized using the ECL plus chemiluminescence kit (PCC-NCI 32132 Amersham, GE Healthcare, Japan) and normalized to β-actin. The Western blots were quantified using ImageJ software.
Ab28767
Manufactured by Abcam
Sourced in United Kingdom
Ab28767 is a primary antibody produced in rabbit. It targets the protein of interest. This product is suitable for use in various immunoassay applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab53161, by Abcam (2 mentions)
Ab37267, by Abcam (1 mentions)
Ab8937, by Abcam (1 mentions)
A2066, by Merck Group (1 mentions)
Anti rabbit or anti mouse igg, by Jackson ImmunoResearch (1 mentions)
Ecl plus chemiluminescence kit, by GE Healthcare (1 mentions)
Aperio digital pathology system, by Leica (1 mentions)
Pgr 1294, by Agilent Technologies (1 mentions)
Ncl l ck5, by Leica (1 mentions)
Immpress peroxidase detection kit, by Vector Laboratories (1 mentions)
A 11056, by Thermo Fisher Scientific (1 mentions)
A21441, by Thermo Fisher Scientific (1 mentions)
Protease inhibitor cocktail, by Roche (1 mentions)
Donkey anti goat igg hrp, by Santa Cruz Biotechnology (1 mentions)
Mouse anti rabbit igg hrp, by Santa Cruz Biotechnology (1 mentions)
Anti mouse igg hrp, by Santa Cruz Biotechnology (1 mentions)
Protease and phosphatase inhibitor cocktail, by Merck Group (1 mentions)
Anti synaptophysin, by Cell Signaling Technology (1 mentions)
Anti psd95, by Cell Signaling Technology (1 mentions)
Anti synapsin, by Abcam (1 mentions)
7 protocols using ab28767
1
Protein Expression Analysis in Cells
2
Immunohistochemistry and ICC Protocol
3
Immunofluorescence Microscopy of FEZ1 and RAR
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HeLa cells plated on a 24‐well dish were fixed and permeabilized with 3.7% formaldehyde and 0.2% Triton X‐100. Cells were incubated in buffer containing 7.5 mg·mL−1 glycine and then blocked with 3% bovine serum albumin and 0.1% Triton X‐100 for 30 min at room temperature. The cells were then incubated with anti‐FEZ1 (1 : 100, Atlas, HPA038490) and anti‐RAR (1 : 100, Abcam, ab28767) primary antibodies for 1 h at room temperature and then with chicken anti‐rabbit 488 (1 : 250, Thermo Fisher, A21441) and donkey anti‐goat 546 (1 : 250, Thermo Fisher, A11056) secondary antibodies for 40 min at room temperature. Nuclei were stained with 1 : 10 000 Hoechst 33342. Slides were examined with Leica confocal microscope.
4
Protein Expression Analysis of BE(2)-C Cells
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The protein of the BE(2)-C cells was extracted using whole cell lysates via a radioimmunoprecipitation assay buffer containing a protease inhibitor cocktail (Roche). The protein samples were separated on a 12%-SDS-polyacrylamide gel electrophoresis plate and detected using antibodies against RARα (ab28767, Abcam), RARβ (sc-552, Santa Cruz), and either β-actin-HRP (KC-5A08, KangChen) or GAPDH-HRP (KC-5G5, KangChen) as an internal control. These samples were then processed with an appropriate HRP-conjugated secondary antibody with regard to the protein of interest. The protein levels were analyzed using ImageJ (Wayne Rasband) and normalized relative to the internal control.
5
Quantification of Apoptosis-Related Proteins
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To quantify the expression levels of RARα, RARβ, RARγ, caspase-3, cleaved caspase-3, and cleaved PARP, cells were lysed with RIPA lysis buffer. Each sample were separated by 8–15% SDS-PAGE, blocked with 5% BSA over night at 4 °C, and incubated with primary antibodies to RARα (1:1000, Abcam, ab28767), RARβ (1:500, Abcam, ab53161), RARγ (1:500, Abcam, ab97569), caspase-3 (1:1000, Cell Signal, 9662S), cleaved caspase-3 (1:1000, Cell Signal, 9664T), or cleaved PARP (1:1000, Cell Signal, 9548T) overnight at 4 °C. Primary antibodies were detected with mouse anti-Rabbit IgG-HRP (1:1000, Santa Cruz, sc-2357), donkey anti-goat IgG-HRP (1:1000, Santa Cruze, sc-2020), or anti-Mouse IgG-HRP (1:1000, Santa Cruz, sc-2005).
6
Western Blot Analysis of Neuroreceptors
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Brain tissue and neurons were lysed in radioimmunoprecipitation assay (RIPA) buffer (50 mM tris, 150 mM NaCl, 1% Triton X-100, and 0.1% SDS) and protease and phosphatase inhibitor cocktail (1:100; Sigma-Aldrich). Western blot was performed as previously described (74 (link)). The primary antibodies stated as follows: anti-RARα (1:500; Abcam, ab28767), anti-ERα (1:1000; Abcam, ab3575), anti-ERβ (1:1000; Santa Cruz Biotechnology, sc-53494), Lamin A/C (1:1000; Sigma-Aldrich, SAB4200236), anti-NMDA receptor 2A (1:250; Abcam, UK), anti-NMDA receptor 1 (1:1000; BD Biosciences, UK), anti-PSD95 (1:1000; Cell Signaling Technology), anti-synapsin (1:1000; Abcam, ab64581), anti-synaptophysin (1:1000; Cell Signaling Technology, #4329), and tubulin (1:10,000; Sigma-Aldrich, T9026). Secondary antibodies anti-rabbit, anti-mouse, and anti-goat IgG were used at 1:10,000 dilution (LI-COR Biosciences, Germany). Immunoreactivity was detected by infrared fluorescence with the LI-COR Odyssey system and quantified with ImageJ software by densitometry analysis of immunoreactive bands.
7
Immunohistochemical Analysis of Fetal Lung and Placenta
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The paraffin-embedded fetal lungs and placentas were sectioned at a thickness of 5 μm, deparaffinized with xylene, and then rehydrated through ethanol and distilled water. Tissue sections were immersed in target retrieval solution (DAKO Ltd, Cambridgeshire, UK) heated for 10 min at 121 °C followed by incubation in 0.03% H 2 O 2 for 30 min to block endogenous peroxidase activity. Sections were incubated overnight at 4 °C with a 1:100 dilution of rabbit polyclonal primary antibody against RXRα (sc553; Santa Cruz Biotechnology, Dallas, TX), 1:50 dilution of goat polyclonal primary antibody against RARα table 7. Primers for quantitative real-time PCR Gene Sequence
(ab28767, Abcam, Cambridge, UK), 1:50 dilution of goat polyclonal primary antibody against RALDH1A3 (sc26713; Santa Cruz Biotechnology), 1:100 dilution of mouse monoclonal primary antibody against CRBP1 (ab24090; Abcam), 1:100 dilution of primary antibody against RARβ (ab53161; Abcam), and 1:100 dilution of mouse monoclonal primary antibody against RBP (sc48384; Santa Cruz Biotechnology). After intensive washing, sections were incubated with corresponding horseradish peroxidase-labeled secondary antibodies and then processed using a DAKO EnVision kit (DAKO), developed with a diaminobenzidine-H 2 O 2 substrate complex, and counterstained with hematoxylin.
(ab28767, Abcam, Cambridge, UK), 1:50 dilution of goat polyclonal primary antibody against RALDH1A3 (sc26713; Santa Cruz Biotechnology), 1:100 dilution of mouse monoclonal primary antibody against CRBP1 (ab24090; Abcam), 1:100 dilution of primary antibody against RARβ (ab53161; Abcam), and 1:100 dilution of mouse monoclonal primary antibody against RBP (sc48384; Santa Cruz Biotechnology). After intensive washing, sections were incubated with corresponding horseradish peroxidase-labeled secondary antibodies and then processed using a DAKO EnVision kit (DAKO), developed with a diaminobenzidine-H 2 O 2 substrate complex, and counterstained with hematoxylin.
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