Western blot analysis was performed as previously described [57 (link)]. The primary antibodies were as follows: RUNX2 (D1L7F) Rabbit mAb #12556(1:1000), APC Antibody #2504(1:1000), β-Catenin (D10A8) XP® Rabbit mAb #8480(1:1000), Cyclin D1 (92G2) Rabbit mAb #2978(1:1000), c-Myc (E5Q6W) Rabbit mAb #18583(1:1000), β-actin (8H10D10) Mouse mAb #3700(1:1000) (All from Cell Signaling Technology, Danvers, MA, USA), anti-ALP (ab83259) (1:1000), anti-osteocalcin (OCN) (ab133612) (1:1000), anti- Osterix (OSX) (ab209484) (1:1000) (All from Abcam, Cambridge, MA, USA). β-actin served as an internal control. Western blot analysis was quantified using ImageJ software (http://rsb.info.nih.gov/ij/ ) and the signal of each target band was normalized to that of the β-actin band.
Anti alp ab83259
Manufactured by Abcam
Sourced in United States
Anti-ALP (ab83259) is a primary antibody that recognizes the Alkaline Phosphatase (ALP) protein. This antibody is intended for use in various research applications to detect and study the ALP protein.
Automatically generated - may contain errors
4 protocols using anti alp ab83259
1
Western Blot Analysis of Osteogenic Markers
2
Quantifying Osteogenic Marker Proteins via Western Blot
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Western blot analysis was performed as previously reported [24 (link)]. The primary antibodies were as follows: anti-ALP (ab83259) (1:1000), anti-osteocalcin (OCN) (ab133612) (1:1000), and anti-Osterix (OSX) (ab209484) (1:1000) (all from Abcam, MA, USA; and RUNX2 (D1L7F) rabbit mAb #12556 (1:1000), APC antibody #2504 (1:1000), 훽-catenin (D10A8) XP Rabbit mAb #8480 (1:1000), and 훽-actin (8H10D10) mouse mAb #3700 (1:1000) (all from Cell Signaling Technology MA, USA). 훽-Actin served as an internal control. Relative densitometry analysis of the Western blot was carried out using ImageJ software. Relative protein levels were quantified as the ratio of the level of target protein to the level of 훽-actin, in each group.
3
In Vivo Bone Formation Assay
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Western blot analysis was performed as previously reported (24) . The primary antibodies were as follows:
anti-ALP (ab83259) (1:1000), anti-osteocalcin (OCN) (ab133612) (1:1000), and anti-Osterix (OSX) (ab209484) (1:1000) (all from Abcam, MA, USA; and RUNX2 (D1L7F) rabbit mAb #12556 (1:1000), APC antibody #2504 (1:1000), 𝛽-catenin (D10A8) XP Rabbit mAb #8480 (1:1000), and 𝛽-actin (8H10D10) mouse mAb #3700 (1:1000) (all from Cell Signaling Technology MA, USA). 𝛽-Actin served as an internal control. Relative densitometry analysis of the Western blot was carried out using ImageJ software.
Relative protein levels were quanti ed as the ratio of the level of target protein to the level of 𝛽-actin, in each group.
In vivo bone formation assay Approximately 10 5 cells (5 × 10 4 HAMSCs and 5 × 10 4 HBMSCs NC /HBMSCs ANRIL / HBMSCs shNC /HBMSCs shANRIL pretreated with LPS) were attached to each HA/TCP biomaterial (Φ 5 × H 2 mm, Sichuan University, Chengdu, Sichuan, China). After 12 hours, the complexes were subcutaneously implanted into the rat mandibular defect area designed as previously reported (four female nude rats per group, with an average weight of 280 g) (23) . All animal experiments were conducted in compliance with the regulations and guidelines of Nanjing Medical University institutional animal care.
anti-ALP (ab83259) (1:1000), anti-osteocalcin (OCN) (ab133612) (1:1000), and anti-Osterix (OSX) (ab209484) (1:1000) (all from Abcam, MA, USA; and RUNX2 (D1L7F) rabbit mAb #12556 (1:1000), APC antibody #2504 (1:1000), 𝛽-catenin (D10A8) XP Rabbit mAb #8480 (1:1000), and 𝛽-actin (8H10D10) mouse mAb #3700 (1:1000) (all from Cell Signaling Technology MA, USA). 𝛽-Actin served as an internal control. Relative densitometry analysis of the Western blot was carried out using ImageJ software.
Relative protein levels were quanti ed as the ratio of the level of target protein to the level of 𝛽-actin, in each group.
In vivo bone formation assay Approximately 10 5 cells (5 × 10 4 HAMSCs and 5 × 10 4 HBMSCs NC /HBMSCs ANRIL / HBMSCs shNC /HBMSCs shANRIL pretreated with LPS) were attached to each HA/TCP biomaterial (Φ 5 × H 2 mm, Sichuan University, Chengdu, Sichuan, China). After 12 hours, the complexes were subcutaneously implanted into the rat mandibular defect area designed as previously reported (four female nude rats per group, with an average weight of 280 g) (23) . All animal experiments were conducted in compliance with the regulations and guidelines of Nanjing Medical University institutional animal care.
4
In Vivo Bone Formation Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Western blot analysis was performed as previously reported (24) . The primary antibodies were as follows:
anti-ALP (ab83259) (1:1000), anti-osteocalcin (OCN) (ab133612) (1:1000), and anti-Osterix (OSX) (ab209484) (1:1000) (all from Abcam, MA, USA; and RUNX2 (D1L7F) rabbit mAb #12556 (1:1000), APC antibody #2504 (1:1000), 𝛽-catenin (D10A8) XP Rabbit mAb #8480 (1:1000), and 𝛽-actin (8H10D10) mouse mAb #3700 (1:1000) (all from Cell Signaling Technology MA, USA). 𝛽-Actin served as an internal control. Relative densitometry analysis of the Western blot was carried out using ImageJ software.
Relative protein levels were quanti ed as the ratio of the level of target protein to the level of 𝛽-actin, in each group.
In vivo bone formation assay Approximately 10 5 cells (5 × 10 4 HAMSCs and 5 × 10 4 HBMSCs NC /HBMSCs ANRIL / HBMSCs shNC /HBMSCs shANRIL pretreated with LPS) were attached to each HA/TCP biomaterial (Φ 5 × H 2 mm, Sichuan University, Chengdu, Sichuan, China). After 12 hours, the complexes were subcutaneously implanted into the rat mandibular defect area designed as previously reported (four female nude rats per group, with an average weight of 280 g) (23) . All animal experiments were conducted in compliance with the regulations and guidelines of Nanjing Medical University institutional animal care.
anti-ALP (ab83259) (1:1000), anti-osteocalcin (OCN) (ab133612) (1:1000), and anti-Osterix (OSX) (ab209484) (1:1000) (all from Abcam, MA, USA; and RUNX2 (D1L7F) rabbit mAb #12556 (1:1000), APC antibody #2504 (1:1000), 𝛽-catenin (D10A8) XP Rabbit mAb #8480 (1:1000), and 𝛽-actin (8H10D10) mouse mAb #3700 (1:1000) (all from Cell Signaling Technology MA, USA). 𝛽-Actin served as an internal control. Relative densitometry analysis of the Western blot was carried out using ImageJ software.
Relative protein levels were quanti ed as the ratio of the level of target protein to the level of 𝛽-actin, in each group.
In vivo bone formation assay Approximately 10 5 cells (5 × 10 4 HAMSCs and 5 × 10 4 HBMSCs NC /HBMSCs ANRIL / HBMSCs shNC /HBMSCs shANRIL pretreated with LPS) were attached to each HA/TCP biomaterial (Φ 5 × H 2 mm, Sichuan University, Chengdu, Sichuan, China). After 12 hours, the complexes were subcutaneously implanted into the rat mandibular defect area designed as previously reported (four female nude rats per group, with an average weight of 280 g) (23) . All animal experiments were conducted in compliance with the regulations and guidelines of Nanjing Medical University institutional animal care.
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