antibody against map1b  (Proteintech)

Journal: Translational Cancer Research

Article Title: Single-patient single-cell RNA sequencing reveals neuroendocrine predominance and immunosuppression in small-cell lung cancer

doi: 10.21037/tcr-2025-1674

Figure Lengend Snippet: Immunochemistry data confirmed the presence of BEX1 and MAP1b in various cell types. (A) Violin plot showing the expression of BEX1 and MAP1b in each cell type. (B) Violin plot showing the expression of BEX1 and MAP1b in tumor tissue compared with adjacent noncancerous tissue. (C,D) Representative immunohistochemical staining of BEX1 (C) and MAP1b (D) in tumor tissue and adjacent noncancerous tissue. *, P<0.05; ***, P<0.001. N, noncancerous tissue; NK, natural killer; T, tumor tissue.

Article Snippet: Slides were subsequently treated at 4 °C overnight with an antibody against BEX1 (Proteintech, Rosemont, IL, USA; Cat#12330-1-AP, 1:100) and an antibody against MAP1b (Proteintech Cat#21633-1-AP, RRID: AB_10793666, 1:400).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Translational Cancer Research

Article Title: Single-patient single-cell RNA sequencing reveals neuroendocrine predominance and immunosuppression in small-cell lung cancer

doi: 10.21037/tcr-2025-1674

Figure Lengend Snippet: MAP1b and BEX1 modulated proliferation, migration and apoptosis in vitro. (A) Levels of MAP1b and BEX1 mRNA in different SCLC cell lines. (B,C) RT-qPCR and Western blotting were applied to examine the knockdown efficiency of MAP1b in NCI-H82 and BEX1 in NCI-H209. (D) A CCK-8 assay was conducted to detect cell proliferation following MAP1b knockdown (left) and BEX1 knockdown (right) in NCI-H82 and NCI-H209. (E) The effect of MAP1b (left) and BEX1 (right) knockdown on the migration capacity of NCI-H82 and NCI-H209. (F-I) The effect of MAP1b (F,G) and BEX1 (H,I) knockdown on cell apoptosis in NCI-H82 and NCI-H209, respectively. Unpaired two-tailed Student’s t -tests and one-way or two-way analysis of variance were used to determine significance. Data are presented as mean ± SD (n=3). **, P<0.01; ****, P<0.0001. CCK-8, Cell Counting Kit-8; OD, optical density; RT-qPCR, reverse transcription quantitative polymerase chain reaction; SCLC, small cell lung cancer; SD, standard deviation.

Article Snippet: Slides were subsequently treated at 4 °C overnight with an antibody against BEX1 (Proteintech, Rosemont, IL, USA; Cat#12330-1-AP, 1:100) and an antibody against MAP1b (Proteintech Cat#21633-1-AP, RRID: AB_10793666, 1:400).

Techniques: Migration, In Vitro, Quantitative RT-PCR, Western Blot, Knockdown, CCK-8 Assay, Two Tailed Test, Cell Counting, Reverse Transcription, Real-time Polymerase Chain Reaction, Standard Deviation

Journal: Translational Cancer Research

Article Title: Single-patient single-cell RNA sequencing reveals neuroendocrine predominance and immunosuppression in small-cell lung cancer

doi: 10.21037/tcr-2025-1674

Figure Lengend Snippet: Immunochemistry data confirmed the presence of BEX1 and MAP1b in various cell types. (A) Violin plot showing the expression of BEX1 and MAP1b in each cell type. (B) Violin plot showing the expression of BEX1 and MAP1b in tumor tissue compared with adjacent noncancerous tissue. (C,D) Representative immunohistochemical staining of BEX1 (C) and MAP1b (D) in tumor tissue and adjacent noncancerous tissue. *, P<0.05; ***, P<0.001. N, noncancerous tissue; NK, natural killer; T, tumor tissue.

Article Snippet: Membranes were incubated on a plate shaker overnight at 4 °C with corresponding primary antibodies against MAP1b (Proteintech Cat#21633-1-AP, RRID: AB_10793666), BEX1 (ThermoFisher Cat#PA5-100404, Thermo Fisher Scientific, Waltham, MA, USA), and Actin (Abcam Cat#ab8226, RRID: AB_306371, Abcam, Cambridge, UK).

Techniques: Expressing, Immunohistochemical staining, Staining

Journal: Translational Cancer Research

Article Title: Single-patient single-cell RNA sequencing reveals neuroendocrine predominance and immunosuppression in small-cell lung cancer

doi: 10.21037/tcr-2025-1674

Figure Lengend Snippet: MAP1b and BEX1 modulated proliferation, migration and apoptosis in vitro. (A) Levels of MAP1b and BEX1 mRNA in different SCLC cell lines. (B,C) RT-qPCR and Western blotting were applied to examine the knockdown efficiency of MAP1b in NCI-H82 and BEX1 in NCI-H209. (D) A CCK-8 assay was conducted to detect cell proliferation following MAP1b knockdown (left) and BEX1 knockdown (right) in NCI-H82 and NCI-H209. (E) The effect of MAP1b (left) and BEX1 (right) knockdown on the migration capacity of NCI-H82 and NCI-H209. (F-I) The effect of MAP1b (F,G) and BEX1 (H,I) knockdown on cell apoptosis in NCI-H82 and NCI-H209, respectively. Unpaired two-tailed Student’s t -tests and one-way or two-way analysis of variance were used to determine significance. Data are presented as mean ± SD (n=3). **, P<0.01; ****, P<0.0001. CCK-8, Cell Counting Kit-8; OD, optical density; RT-qPCR, reverse transcription quantitative polymerase chain reaction; SCLC, small cell lung cancer; SD, standard deviation.

Article Snippet: Membranes were incubated on a plate shaker overnight at 4 °C with corresponding primary antibodies against MAP1b (Proteintech Cat#21633-1-AP, RRID: AB_10793666), BEX1 (ThermoFisher Cat#PA5-100404, Thermo Fisher Scientific, Waltham, MA, USA), and Actin (Abcam Cat#ab8226, RRID: AB_306371, Abcam, Cambridge, UK).

Techniques: Migration, In Vitro, Quantitative RT-PCR, Western Blot, Knockdown, CCK-8 Assay, Two Tailed Test, Cell Counting, Reverse Transcription, Real-time Polymerase Chain Reaction, Standard Deviation

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: Cell death & disease

Article Title: Opposing roles for GSK3β and ERK1-dependent phosphorylation of huntingtin during neuronal dysfunction and cell death in Huntington's disease.

doi: 10.1038/s41419-025-07524-0

Figure Lengend Snippet: Fig. 1 The proteomic network on HTT containing light membranes is dramatically altered in HD iPSC-derived neurons. A Representative images from healthy/normal (WT, Q17) or diseased (HD, Q109) human iPSCs stained with the pluripotent marker OCT-4, the neuronal precursor (NPC) marker Nestin and the mature neuronal markers MAP2 and βIII-Tubulin. Hoechst stains nuclei. Scale = 25 μm. Differentiated neurons show Synaptophysin (SYP) positive staining. Scale = 10 μm. B Electrophysiological analysis of WT and HD human neurons differentiated from iPSCs show action potentials, which are abolished in the presence of TTX or TEA. C Schematic diagram of human iNeuron lysate fractionation into perinuclear supernatant (PNS), light membrane (LM), soluble (SF), and heavy membrane (P1) fractions by ultra- centrifugation and sucrose gradient separation. D Workflow for quality control and quantification of unique peptides identified from LC-MS of HTT-IPs from WT or HD human iNeurons. E Hierarchical cluster heat map showing the avg. relative abundance (spectral count; SpC) of 800 proteins (≥3 unique peptides/trial across ≥2 biological replicates) quantified across the WT and HD HTT-IPs with a normalized fold change (FC) threshold of ±2X and a significance threshold of p < 0.05 determined by a Welch’s t test across three independent biological replicates. Increased in HD HTT-IP = red, decreased in HD HTT-IP = blue. In addition, proteins were identified in only WT HTT-IP (lost = green) or in only HD HTT-IP (gained = orange). F Volcano plot with the y axis depicting significance (−log10[p value]) and the x axis depicting fold change of individual peptides between HD and WT HTT-IPs (log2[FC]). Three independent biological replicates were performed for each genotype. A negative, no-antibody IP was performed to account for non-specific peptide association with magnetic beads. G Representative western blot of HTT-IP from WT or HD LMs, probed against HTT, KIF5A, KIF5B, KIF5C, DNCT, MAP1B, MAP2, RAB2, RAB5, RAB7, VPS35, or SUMO2. Except for KIF5A, all show presence in WT and HD HTT-IP. No bands are seen in the negative no antibody control (−Crtl). n = 3. Statistical analysis was conducted using the two-sample two-sided Student’s t test comparing signal/noise intensity between bands in WT and HD conditions normalized to WT. Data represented as mean ± SEM. ns = p > 0.05, *p < 0.05, **p < 0.005.

Article Snippet: Blots were blocked using TBST with 5% BSA for 60 mins at 25 °C and incubated with primary antibodies (SYT1 (Thermofisher 1:1000), Rab4 (Abcam 1:1000), Rab5 (Abcam 1:1000), Rab2 (SCBT 1:500), Rab7 (SCBT 1:500), VPS35 (SCBT 1:500), SUMO2/3 (Cytoskeleton 1:500), KIF5A (Goldstein 1:250), KIF5B (Goldstein 1:250), KIF5C (Goldstein 1:250), DIC (Abcam 1:1000), DNCT (Abcam 1:1000), Actin (ThermoFisher 1:1000), Tubulin (Abcam 1:2000), HTT rabbit polyclonal (Abcam 1:1000), HTT mouse monoclonal (EMD Millipore 1:1000), Golgi (Millipore Sigma 1:1000), Cytochrome C (Santa Cruz 1:1000), TOM20 (CellSignaling Technology 1:500), MAP1B (SCBT 1:1000), MAP2 (BD Pharmigen 1:1000), Total AKT1 (CellSignaling Technology 1:1000), pAKT1 (Ser473, CellSignaling Technology 1:1000), Total GSK3α/β (CellSignaling Technology 1:1000), pGSK3α/β (pY279/pY216; Abcam 1:1000), or ERK (pan-ERK; BD Transduction Laboratories 1:1000) for 16 h at 4 °C.

Techniques: Derivative Assay, Staining, Marker, Fractionation, Membrane, Centrifugation, Control, Liquid Chromatography with Mass Spectroscopy, Magnetic Beads, Western Blot