rabbit polyclonal antibodies against total map1b  (Cell Signaling Technology Inc)

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Intermediate Filament Protein BFSP1 Maintains Oocyte Asymmetric Division by Modulating Spindle Length.

doi: 10.1002/advs.202504066

Figure Lengend Snippet: Figure 4. BFSP1 interacts with MAP1B and affects its protein stability. A) Identification of BFSP1 binding proteins by IP/MS analysis. Protein name, cov- erage percentage, the number of identified peptides, and molecular weight were shown in the table. B) Representation of the 3D structure and predicted interaction of mouse BFSP1 and MAP1B using AlphaFold databank by HDOCK server. C) Co-IP using anti-BFSP1 antibody followed by immunoblotting analysis with anti-MAP1B and anti-BFSP1 antibodies. D) Co-IP using anti-MAP1B antibody followed by immunoblotting analysis with anti-BFSP1 and anti-MAP1B antibodies. E) Representative images of MAP1B in control and BFSP1-KD oocytes. Scale bar, 10 μm. F) The ratio of MAP1B fluorescence intensity in the spindle region to the cytoplasmic region was measured in control and BFSP1-KD oocytes. G) Protein levels of MAP1B in control, BFSP1- KD, and BFSP1-rescue oocytes as assessed by immunoblotting analysis. The band intensity of BFSP1 and MAP1B was normalized with that of GAPDH. H) The band intensities of BFSP1 and MAP1B in the blots were normalized with that of GAPDH. Data in (F) were expressed as mean ± SD, and (H) were expressed as mean ± SEM of at least three independent experiments. ***P < 0.001; ns, no significance.

Article Snippet: Antibodies: Rabbit polyclonal anti-BFSP1 antibody (Cat# A3764) and rabbit monoclonal anti-Myc antibody (Cat# AE070) were purchased fromAbclonal (Wuhan, China); rabbit polyclonal anti-MAP1B antibody (Cat# 21633-1-AP), rabbit polyclonal antiHSP90α antibody (Cat# 13171-1-AP), mouse monoclonal anti-βActin antibody (Cat# 66009-1-lg), rabbit polyclonal anti-HA antibody (Cat# 51064-2-AP), and mouse monoclonal anti-GAPDH antibody (Cat# 60004-1-lg) were purchased from Proteintech (Rosemont, IL, USA); mouse monoclonal anti-α-Tubulin-FITC antibody (Cat# F2168) was purchased from Sigma–Aldrich (St. Louis, MO, USA); rabbit monoclonal anti-Vinculin antibody (Cat# CY5164) was purchased from Always (Shanghai, China).

Techniques: Binding Assay, Protein-Protein interactions, Molecular Weight, Co-Immunoprecipitation Assay, Western Blot, Control

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Intermediate Filament Protein BFSP1 Maintains Oocyte Asymmetric Division by Modulating Spindle Length.

doi: 10.1002/advs.202504066

Figure Lengend Snippet: Figure 5. MAP1B depletion impairs the oocyte meiotic maturation and spindle length control. A) Representative images of oocytes at M II stage in control and MAP1B-KD groups. Yellow asterisks indicate oocytes that failed to extrude the first polar body, and red asterisks indicate oocytes with symmetric division. Scale bar, 80 μm. B) The GVBD rate was quantified in control (n = 202) and MAP1B-KD (n = 189) oocytes. C) The PBE rate was quantified in control (n = 202) and MAP1B-KD (n = 189) oocytes. D) The rate of symmetric division was quantified in control (n = 202) and MAP1B-KD (n = 189) oocytes. E) Representative images of spindle length in control and MAP1B-KD oocytes at M I stage. Oocytes were immunostained for 𝛼-tubulin and 𝛾-tubulin. Scale bar, 15 μm. F) Spindle length was measured between two spindle poles in control (n = 23) and MAP1B-KD (n = 26) oocytes at M I stage. G) Representative images of spindle length in control and MAP1B-KD oocytes at AT I stage. Oocytes were immunostained for 𝛼-tubulin and 𝛾-tubulin. Scale bar, 15 μm. H) Spindle length was measured between two spindle poles in control (n = 15) and MAP1B-KD (n = 19) oocytes at AT I stage. Data in (B), (C), and (D) were expressed as mean ± SEM, and (F) and (H) were expressed as mean ± SD of at least three independent experiments. *P < 0.05; **P < 0.01; ***P < 0.001.

Article Snippet: Antibodies: Rabbit polyclonal anti-BFSP1 antibody (Cat# A3764) and rabbit monoclonal anti-Myc antibody (Cat# AE070) were purchased fromAbclonal (Wuhan, China); rabbit polyclonal anti-MAP1B antibody (Cat# 21633-1-AP), rabbit polyclonal antiHSP90α antibody (Cat# 13171-1-AP), mouse monoclonal anti-βActin antibody (Cat# 66009-1-lg), rabbit polyclonal anti-HA antibody (Cat# 51064-2-AP), and mouse monoclonal anti-GAPDH antibody (Cat# 60004-1-lg) were purchased from Proteintech (Rosemont, IL, USA); mouse monoclonal anti-α-Tubulin-FITC antibody (Cat# F2168) was purchased from Sigma–Aldrich (St. Louis, MO, USA); rabbit monoclonal anti-Vinculin antibody (Cat# CY5164) was purchased from Always (Shanghai, China).

Techniques: Control

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Intermediate Filament Protein BFSP1 Maintains Oocyte Asymmetric Division by Modulating Spindle Length.

doi: 10.1002/advs.202504066

Figure Lengend Snippet: Figure 6. Restored MAP1B protein levels mitigate the meiotic defects in- duced in BFSP1 depleted-oocytes. A) Representative images of oocytes at M II stage in control, BFSP1-KD, and MAP1B-rescue groups. For the res- cue experiment, MAP1B-EGFP mRNA was microinjected to GV oocytes 20 h after microinjection of BFSP1 siRNAs. Yellow asterisks indicate oocytes that failed to extrude the first polar body, and red asterisks indicate oocytes with symmetric division. Scale bar, 80 μm. B) The GVBD rate was quan- tified in control (n = 180), BFSP1-KD (n = 174), and MAP1B-rescue (n = 185) oocytes. C) The PBE rate was quantified in control (n = 180), BFSP1- KD (n = 174), and MAP1B-rescue (n = 185) oocytes. D) The rate of sym- metric division was quantified in control (n = 180), BFSP1-KD (n = 174), and MAP1B-rescue (n = 185) oocytes. E) Representative images of spin- dle length in control, BFSP1-KD, and MAP1B-rescue oocytes at M I stage. Oocytes were immunostained for 𝛼-tubulin and 𝛾-tubulin. Scale bar, 15 μm. F) Spindle length was measured between two spindle poles in control (n = 19), BFSP1-KD (n = 19), and MAP1B-rescue (n = 14) oocytes at M I stage. G) Representative images of spindle length in control, BFSP1-KD, and MAP1B-rescue oocytes at AT I stage. Oocytes were immunostained

Article Snippet: Antibodies: Rabbit polyclonal anti-BFSP1 antibody (Cat# A3764) and rabbit monoclonal anti-Myc antibody (Cat# AE070) were purchased fromAbclonal (Wuhan, China); rabbit polyclonal anti-MAP1B antibody (Cat# 21633-1-AP), rabbit polyclonal antiHSP90α antibody (Cat# 13171-1-AP), mouse monoclonal anti-βActin antibody (Cat# 66009-1-lg), rabbit polyclonal anti-HA antibody (Cat# 51064-2-AP), and mouse monoclonal anti-GAPDH antibody (Cat# 60004-1-lg) were purchased from Proteintech (Rosemont, IL, USA); mouse monoclonal anti-α-Tubulin-FITC antibody (Cat# F2168) was purchased from Sigma–Aldrich (St. Louis, MO, USA); rabbit monoclonal anti-Vinculin antibody (Cat# CY5164) was purchased from Always (Shanghai, China).

Techniques: Control, Microinjection

Journal: Advanced science (Weinheim, Baden-Wurttemberg, Germany)

Article Title: Intermediate Filament Protein BFSP1 Maintains Oocyte Asymmetric Division by Modulating Spindle Length.

doi: 10.1002/advs.202504066

Figure Lengend Snippet: Figure 7. BFSP1 maintains MAP1B protein levels by recruiting HSP90𝛼. A) Co-IP using anti-BFSP1 antibody followed by immunoblotting analysis with anti-HSP90𝛼and anti-BFSP1 antibodies. B) Protein levels of MAP1B in control and 17-AAG-treated oocytes as assessed by immunoblotting analysis. C) The band intensity of MAP1B in the blots was normalized with that of 𝛽-Actin. D) Protein levels of HSP90𝛼in control and BFSP1-KD oocytes as assessed by immunoblotting analysis. E) The band intensities of BFSP1 and HSP90𝛼in the blots were normalized with that of 𝛽-Actin. F) Representative images of HSP90𝛼localization in the spindle region in control and BFSP1-KD oocytes. Scale bars, 20 μm, 10 μm. Data in (C) and (E) were expressed as mean ± SEM of at least three independent experiments. ***P < 0.001; ns, no significance.

Article Snippet: Antibodies: Rabbit polyclonal anti-BFSP1 antibody (Cat# A3764) and rabbit monoclonal anti-Myc antibody (Cat# AE070) were purchased fromAbclonal (Wuhan, China); rabbit polyclonal anti-MAP1B antibody (Cat# 21633-1-AP), rabbit polyclonal antiHSP90α antibody (Cat# 13171-1-AP), mouse monoclonal anti-βActin antibody (Cat# 66009-1-lg), rabbit polyclonal anti-HA antibody (Cat# 51064-2-AP), and mouse monoclonal anti-GAPDH antibody (Cat# 60004-1-lg) were purchased from Proteintech (Rosemont, IL, USA); mouse monoclonal anti-α-Tubulin-FITC antibody (Cat# F2168) was purchased from Sigma–Aldrich (St. Louis, MO, USA); rabbit monoclonal anti-Vinculin antibody (Cat# CY5164) was purchased from Always (Shanghai, China).

Techniques: Co-Immunoprecipitation Assay, Western Blot, Control

Journal: iScience

Article Title: Single-cell atlas unveils cellular heterogeneity and novel markers in human neonatal and adult intervertebral discs

doi: 10.1016/j.isci.2022.104504

Figure Lengend Snippet:

Article Snippet: Rabbit Polyclonal Anti-MAP1B , Novus Biologicals , NBP3-04801-20ul.

Techniques: Recombinant, Gene Expression, Software

Journal: PLoS ONE

Article Title: Repulsive Axon Guidance by Draxin Is Mediated by Protein Kinase B (Akt), Glycogen Synthase Kinase-3β (GSK-3β) and Microtubule-Associated Protein 1B

doi: 10.1371/journal.pone.0119524

Figure Lengend Snippet: A. Cortical explants from newborn wild-type or MAP1B-/- mice, cultured for 48 h in the presence or absence (control) of 10 nM draxin or 100 ng/ml semaphorin 3A (Sema3A). Scale bar = 100 μm. B. Quantification of the longest neurites of 30 explants in 3 independent experiments. C. Growth cones of cortical neurons from newborn wild-type or MAP1B-/- mice cultured for 60 h, treated for 1 h with 100 nM draxin or 30 min with 100 ng/ml semaphorin 3A (Sema3A), fixed and stained for F-actin. Scale bar = 10 μm. D. Percentage of collapsed growth cones. For each experimental condition the growth cones of 30 neurons in each of 3 independent experiments were evaluated.

Article Snippet: Primary antibodies: mouse monoclonal antibodies against phospho-MAP1B (SMI31; 1:1000; Covance) and neurofilament H (1:1000; Cell Signaling); rabbit polyclonal antibodies against total-MAP1B raised against peptides ATVVVEATEPEPSGC and ETVTEEHLRRAIGN [ ], phospho Ser473 Akt (1:1000; Cell Signaling), total Akt (1:1000; Cell Signaling) and GAPDH (1:3000; Sigma-Aldrich); rabbit monoclonal antibodies against phospho Ser9-GSK-3β (1:1000; Cell Signaling) and total GSK-3β (1:1000; Cell Signaling).

Techniques: Cell Culture, Control, Staining

Journal: PLoS ONE

Article Title: Repulsive Axon Guidance by Draxin Is Mediated by Protein Kinase B (Akt), Glycogen Synthase Kinase-3β (GSK-3β) and Microtubule-Associated Protein 1B

doi: 10.1371/journal.pone.0119524

Figure Lengend Snippet: Immunoblot analyses of cortical neurons from newborn wild-type mice cultured for 60 h, treated with draxin in the absence or presence of the GSK-3β inhibitor SB216763 for the indicated times, lysed and probed using the indicated antibodies. A. Representative immunoblot for determination of MAP1B phosphorylation in response to draxin treatment. B. The relative level of phosphorylated MAP1B (P-MAP1B) was determined by normalizing the P-MAP1B signal to the signal for total MAP1B 30 min after draxin addition in 3 independent experiments. C. Representative immunoblot for determination of MAP1B protein levels in response to draxin treatment. D. The relative level of MAP1B was determined by normalizing the MAP1B signal to the signal for neurofilament H in 3 independent experiments. Draxin treatment did not lead to a significant change in MAP1B levels.

Article Snippet: Primary antibodies: mouse monoclonal antibodies against phospho-MAP1B (SMI31; 1:1000; Covance) and neurofilament H (1:1000; Cell Signaling); rabbit polyclonal antibodies against total-MAP1B raised against peptides ATVVVEATEPEPSGC and ETVTEEHLRRAIGN [ ], phospho Ser473 Akt (1:1000; Cell Signaling), total Akt (1:1000; Cell Signaling) and GAPDH (1:3000; Sigma-Aldrich); rabbit monoclonal antibodies against phospho Ser9-GSK-3β (1:1000; Cell Signaling) and total GSK-3β (1:1000; Cell Signaling).

Techniques: Western Blot, Cell Culture, Phospho-proteomics

Journal: PLoS ONE

Article Title: Repulsive Axon Guidance by Draxin Is Mediated by Protein Kinase B (Akt), Glycogen Synthase Kinase-3β (GSK-3β) and Microtubule-Associated Protein 1B

doi: 10.1371/journal.pone.0119524

Figure Lengend Snippet: Immunoblot analyses of cortical neurons from newborn wild-type ( A ) or MAP1B-/- ( B ) mice cultured for 60 h, treated with draxin for the indicated times, lysed and probed using the indicated antibodies. The GSK-3β doublets represent the GSK-3β1 and GSK-3β2 isoforms .The relative levels of GSK-3β phosphorylated at Ser9 (P-GSK-3β) and Akt phosphorylated on Ser473 (P-Akt) were determined by normalizing the signals for the phosphorylated proteins to the corresponding signals for the total proteins in 3 independent experiments.

Article Snippet: Primary antibodies: mouse monoclonal antibodies against phospho-MAP1B (SMI31; 1:1000; Covance) and neurofilament H (1:1000; Cell Signaling); rabbit polyclonal antibodies against total-MAP1B raised against peptides ATVVVEATEPEPSGC and ETVTEEHLRRAIGN [ ], phospho Ser473 Akt (1:1000; Cell Signaling), total Akt (1:1000; Cell Signaling) and GAPDH (1:3000; Sigma-Aldrich); rabbit monoclonal antibodies against phospho Ser9-GSK-3β (1:1000; Cell Signaling) and total GSK-3β (1:1000; Cell Signaling).

Techniques: Western Blot, Cell Culture

Journal: PLoS ONE

Article Title: Repulsive Axon Guidance by Draxin Is Mediated by Protein Kinase B (Akt), Glycogen Synthase Kinase-3β (GSK-3β) and Microtubule-Associated Protein 1B

doi: 10.1371/journal.pone.0119524

Figure Lengend Snippet: Draxin interaction with DCC triggers inactivation of the Akt pathway. This relieves GSK-3β from Akt-mediated inhibition leading to an increase in phosphorylation of MAP1B and reconfiguration of the cytoskeleton to promote growth cone collapse and inhibition of neurite extension.

Article Snippet: Primary antibodies: mouse monoclonal antibodies against phospho-MAP1B (SMI31; 1:1000; Covance) and neurofilament H (1:1000; Cell Signaling); rabbit polyclonal antibodies against total-MAP1B raised against peptides ATVVVEATEPEPSGC and ETVTEEHLRRAIGN [ ], phospho Ser473 Akt (1:1000; Cell Signaling), total Akt (1:1000; Cell Signaling) and GAPDH (1:3000; Sigma-Aldrich); rabbit monoclonal antibodies against phospho Ser9-GSK-3β (1:1000; Cell Signaling) and total GSK-3β (1:1000; Cell Signaling).

Techniques: Inhibition, Phospho-proteomics