Prior to the histology analysis, rat DRG tissues were put into reaction with 4% paraformaldehyde and 30% sucrose solution for perfusion. Frozen slices of DRG tissues were then treated with primary antibodies against CDMP1 (1:500; cat. no. ab93855; Abcam, Cambridge, UK) at 4°C for 1 h, and blocked with blocking reagent (PBS-Triton X-100 and 0.5% bovine serum albumin (BSA; Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) at room temperature for 60 min. The tissue slices were then incubated with horseradish peroxidase-conjugated secondary antibodies (goat anti-rabbit; 1:1,000; cat. no. ab150077; Abcam) at room temperature for 90 min. The tissue samples were observed by fluorescence microscope (magnification, ×100) after being treated with diaminobenzidine (Sigma-Aldrich; Merck KGaA, Darmstadt, Germany).
Ab93855
Manufactured by Abcam
Sourced in United States
Ab93855 is a recombinant antibody produced in HEK293 cells. It is specific for the protein target X and can be used for various research applications.
Automatically generated - may contain errors
Lab products found in correlation
Ab30455, by Abcam (2 mentions)
Ab39670, by Abcam (2 mentions)
Ab150077, by Abcam (1 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Beyotime (1 mentions)
Ab93876, by Abcam (1 mentions)
Ab22552, by Abcam (1 mentions)
Ripa lysis buffer, by Beyotime (1 mentions)
Enhanced chemi luminescence (ecl), by Merck Group (1 mentions)
Ab95462, by Abcam (1 mentions)
Ab23981, by Abcam (1 mentions)
Ab8448, by Abcam (1 mentions)
Ab9485, by Abcam (1 mentions)
Bca protein assay kit, by Solarbio (1 mentions)
Pvdf membrane, by Merck Group (1 mentions)
Ripa lysate, by Beyotime (1 mentions)
Anti β actin antibody, by Proteintech (1 mentions)
Ecl chromogenic agent, by Merck Group (1 mentions)
Ab26350, by Santa Cruz Biotechnology (1 mentions)
Ab226821, by Abcam (1 mentions)
Rm2235, by Leica (1 mentions)
5 protocols using ab93855
1
CDMP1 Immunohistochemistry in Rat DRG
2
Western Blot Analysis of Osteogenic Markers
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Cell lysates were obtained using RIPA lysis buffer (Beyotime, Shanghai, China) containing 10 mM phenylmethylsulphonylfluoride as a protease inhibitor (PMSF; Beyotime) and 50 µg of total protein was separated in a Bis-Tris polyacrylamide gel and transferred onto a nitrocellulose membrane. The membrane was then incubated in 5% bovine serum albumin (BSA) containing primary rabbit-anti-human polyclonal antibodies at 4 °C overnight. After incubating with horseradish peroxidase (HRP)-conjugated goat-anti-rabbit antibody at room for 1 h, protein was detected using electrochemiluminescence (ECL; Millipore, Darmstadt, Germany). The following primary rabbit-anti-human antibodies were used: anti-FOXO1 (1:1000; ab39670, Abcam, Cambridge, MA, USA); anti-GDF5 (1:1000; ab93855, Abcam); anti-SOX6 (1:1000; ab30455, Abcam); anti-Runx2 (1:1000; ab23981, Abcam); anti-Sp7/Osterix (1:2000; ab22552, Abcam); anti-ALP (1:2000; ab95462, Abcam); anti-OCN (1:500; ab93876, Abcam); anti-OPN (1:1000; ab8448, Abcam); and anti-GAPDH (1:2500; ab9485, Abcam). The results of Western blots were quantified using image-J software (http://imagej.net ) analysis.
3
Multi-Staining and Immunohistochemistry of Musculoskeletal Tissues
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Serial 5-mm-thick sections were prepared from paraffin-embedded specimens for staining.
Hematoxylin-eosin staining was performed in an autostainer machine (ST5010 XL; Leica Microsystems, Mannheim, Germany) using standard procedures.
Elastic Fibers staining kit for ligament, Alcian Blue staining kit for cartilage and Fast Green staining kit for bone were purchased from Leagene Biotech (DC0066; DB0060; DZ0046).
Sections for immunohistochemical staining were carried out as described previously [49 (link)]. The primary rabbit anti-human antibodies were: anti-FOXO1 (1:200; ab39670, Abcam); anti-GDF5 (1:200; ab93855, Abcam); anti-SOX6 (1:200; ab30455, Abcam).
Hematoxylin-eosin staining was performed in an autostainer machine (ST5010 XL; Leica Microsystems, Mannheim, Germany) using standard procedures.
Elastic Fibers staining kit for ligament, Alcian Blue staining kit for cartilage and Fast Green staining kit for bone were purchased from Leagene Biotech (DC0066; DB0060; DZ0046).
Sections for immunohistochemical staining were carried out as described previously [49 (link)]. The primary rabbit anti-human antibodies were: anti-FOXO1 (1:200; ab39670, Abcam); anti-GDF5 (1:200; ab93855, Abcam); anti-SOX6 (1:200; ab30455, Abcam).
4
Protein Expression Analysis of ADSCs
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The ADSCs were washed by cold PBS 3 times, then 150 μL RIPA lysate (Beyotime Biotechnology, Shanghai, China) was added to extract total protein. The cells were lysed in ice water by ultrasound, and the protein content was determined by the BCA method (BCA Protein Assay Kit, Solarbio, Beijing, China). An equal amount of proteins were taken from each group for 10% SDS-PAGE, and the proteins on the gel were transferred to PVDF membranes (Millipore, Bedford, MA, USA). The membranes were blocked at 4 °C for 1 h and then incubated at 4 °C overnight with the following primary antibodies (concentration: 1:1000): the BMP-14 antibody (ab93855, Abcam, USA), anti-ACAN antibody (13880-1-AP, Proteintech, Wuhan, China), Anti-collagen II antibody (28459-1-AP, Proteintech, Wuhan, China), anti-SOX9 antibody (67439, Proteintech, Wuhan, China) and anti-β-actin antibody (66009, Proteintech, Wuhan, China). After being cleaned twice with TBST, the membranes were incubated at room temperature for 1 h with fluorescein-labeled Goat anti-Rabbit IgG (ab205718, 1:2000). The antibodies were all from Abcam (Cambridge, UK). Finally, the membranes were cleaned three times, exposed with the ECL chromogenic agent (Millipore, Bedford, MA, USA), and imaged with an automatic developer (ChemiDoc XRS imaging system).
5
Immunohistochemical Analysis of Testicular Tissue
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Human seminiferous tubules were fixed with 4% paraformaldehyde and stored in 70% ethanol. Then, fixed samples were embedded in paraffin and sliced into 5-μm sections by microtome (Leica RM2235). Immunostaining was performed as described previously (42 (link)). Deparaffined and rehydrated sections were washed in PBS for 5 min after antigen retrieval and then blocked with 5% BSA at room temperature for 1 h. Subsequently, the sections were incubated with primary antibodies at 4°C for more than 8 hours and secondary antibodies (Jackson ImmunoResearch, Alexa 488-, Alexa 594-) for 1 h at room temperature. Nuclei were counterstained with 10 μg/ml Hoechst 33342 for 15 min at room temperature. All images were captured with a ZEISS LSM880 confocal microscope. The experiments performed in this study were approved by the Third Affiliated Hospital of Guangzhou Medical University (2017-055). And informed consents to every donor in our study have been signed.
The following primary antibodies were used: mouse anti-γH2AX (Abcam, ab26350), mouse anti-FGFR3 (Santa Cruz, sc-13121), mouse anti-BMPR1B (Abcam, ab1565836), mouse anti-DDX4 (Abcam, ab27591), rabbit anti-INHBB (Abcam, ab69286), mouse anti-ACVR2B (R&D Systems, MAB3392), rabbit anti-pSMAD2 (Cell Signaling Technology, #18338), rabbit anti-GDF5 (Abcam, ab93855), rabbit anti-pSMAD1 (Abcam, ab226821) and rabbit anti-ACVR1B (Abcam, ab109300).
The following primary antibodies were used: mouse anti-γH2AX (Abcam, ab26350), mouse anti-FGFR3 (Santa Cruz, sc-13121), mouse anti-BMPR1B (Abcam, ab1565836), mouse anti-DDX4 (Abcam, ab27591), rabbit anti-INHBB (Abcam, ab69286), mouse anti-ACVR2B (R&D Systems, MAB3392), rabbit anti-pSMAD2 (Cell Signaling Technology, #18338), rabbit anti-GDF5 (Abcam, ab93855), rabbit anti-pSMAD1 (Abcam, ab226821) and rabbit anti-ACVR1B (Abcam, ab109300).
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