Anti-Malectin/MLEC Antibody Picoband®

Western blot analysis of Malectin/MLEC using anti-Malectin/MLEC antibody (A10455-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: human RT4 whole cell lysates,
Lane 2: human THP-1 whole cell lysates,
Lane 3: human A549 whole cell lysates,
Lane 4: human SIHA whole cell lysates,
Lane 5: human A431 whole cell lysates,
Lane 6: human MCF-7 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Malectin/MLEC antigen affinity purified polyclonal antibody (Catalog # A10455-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Malectin/MLEC at approximately 32 kDa. The expected band size for Malectin/MLEC is at 32 kDa.

Western blot analysis of Malectin/MLEC using anti-Malectin/MLEC antibody (A10455-1).
Electrophoresis was performed on a 5-20% SDS-PAGE gel at 70V (Stacking gel) / 90V (Resolving gel) for 2-3 hours. The sample well of each lane was loaded with 30 ug of sample under reducing conditions.
Lane 1: rat liver tissue lysates,
Lane 2: mouse liver tissue lysates,
Lane 3: mouse NIH/3T3 whole cell lysates.
After electrophoresis, proteins were transferred to a nitrocellulose membrane at 150 mA for 50-90 minutes. Blocked the membrane with 5% non-fat milk/TBS for 1.5 hour at RT. The membrane was incubated with rabbit anti-Malectin/MLEC antigen affinity purified polyclonal antibody (Catalog # A10455-1) at 0.5 μg/mL overnight at 4°C, then washed with TBS-0.1%Tween 3 times with 5 minutes each and probed with a goat anti-rabbit IgG-HRP secondary antibody at a dilution of 1:5000 for 1.5 hour at RT. The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit (Catalog # EK1002) with Tanon 5200 system. A specific band was detected for Malectin/MLEC at approximately 32 kDa. The expected band size for Malectin/MLEC is at 32 kDa.

IHC analysis of Malectin/MLEC using anti-Malectin/MLEC antibody (A10455-1).
Malectin/MLEC was detected in a paraffin-embedded section of human bladder urothelial carcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Malectin/MLEC Antibody (A10455-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Malectin/MLEC using anti-Malectin/MLEC antibody (A10455-1).
Malectin/MLEC was detected in a paraffin-embedded section of human colorectal adenocarcinoma tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Malectin/MLEC Antibody (A10455-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Malectin/MLEC using anti-Malectin/MLEC antibody (A10455-1).
Malectin/MLEC was detected in a paraffin-embedded section of human liver cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Malectin/MLEC Antibody (A10455-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Malectin/MLEC using anti-Malectin/MLEC antibody (A10455-1).
Malectin/MLEC was detected in a paraffin-embedded section of human lung cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Malectin/MLEC Antibody (A10455-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Malectin/MLEC using anti-Malectin/MLEC antibody (A10455-1).
Malectin/MLEC was detected in a paraffin-embedded section of human spleen tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Malectin/MLEC Antibody (A10455-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Malectin/MLEC using anti-Malectin/MLEC antibody (A10455-1).
Malectin/MLEC was detected in a paraffin-embedded section of human testicular germ cell tumors tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Malectin/MLEC Antibody (A10455-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Malectin/MLEC using anti-Malectin/MLEC antibody (A10455-1).
Malectin/MLEC was detected in a paraffin-embedded section of human thyroid cancer tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Malectin/MLEC Antibody (A10455-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

IHC analysis of Malectin/MLEC using anti-Malectin/MLEC antibody (A10455-1).
Malectin/MLEC was detected in a paraffin-embedded section of rat kidney tissue. Heat mediated antigen retrieval was performed in EDTA buffer (pH 8.0, epitope retrieval solution). The tissue section was blocked with 10% goat serum. The tissue section was then incubated with 2 μg/ml rabbit anti-Malectin/MLEC Antibody (A10455-1) overnight at 4°C. Peroxidase Conjugated Goat Anti-rabbit IgG was used as secondary antibody and incubated for 30 minutes at 37°C. The tissue section was developed using HRP Conjugated Rabbit IgG Super Vision Assay Kit (Catalog # SV0002) with DAB as the chromogen.

Flow Cytometry analysis of SiHa cells using anti-Malectin/MLEC antibody (A10455-1).
Overlay histogram showing SiHa cells stained with A10455-1 (Blue line). To facilitate intracellular staining, cells were fixed with 4% paraformaldehyde and permeabilized with permeabilization buffer. The cells were blocked with 10% normal goat serum. And then incubated with rabbit anti-Malectin/MLEC Antibody (A10455-1, 1 μg/1x10⁶ cells) for 30 min at 20°C. DyLight®488 conjugated goat anti-rabbit IgG (BA1127, 5-10 μg/1x10⁶ cells) was used as secondary antibody for 30 minutes at 20°C. Isotype control antibody (Green line) was rabbit IgG (1 μg/1x10⁶) used under the same conditions. Unlabelled sample (Red line) was also used as a control.

TUSC3 promotes GCS2-mediated glucose trimming and prevents insoluble G2M9-BMP4 accumulation Show full caption (A) Anti-HA and anti-OS9 immunoblots of MEFs cultured at 20°C and treated with DTT (6 h), NBDNJ (2 mM, 16 h), or DMSO (control). The five BMP4-HA species are marked (top to bottom: G3M9-BMP4, G2M9-BMP4, G1M9-BMP4, M9-BMP4, and nonglycosylated/deglycosylated BMP4; red arrow). Tubulin served as a loading control. (B) Anti-HA immunoblots of MEFs transfected with CTRL or Tusc3 siRNA and treated with NBDNJ (8 h), GAA (50 mU/mL, 1 h at 37°C), or NMS-873 (10 μM, 6 h). (C) Malectin far-western blot of total protein lysates from MEFs under the indicated conditions. (D) HA IP of BMP4-HA from control and Tusc3 -knockdown MEFs, followed by malectin far-western blot. (E) Co-IP of BMP4-HA-Myc with endogenous malectin in MEFs expressing BMP4-HA-Myc, cotransfected with control or Tusc3 siRNA, and treated with DMSO or DTT. Cell lysates were subjected to anti-HA IP, and both input lysates and IP fractions were immunoblotted with the indicated antibodies. (F) Western blot analysis of BMP4-HA in soluble and insoluble protein fractions from control and Tusc3 -knockdown MEFs. Representative of two independent experiments is shown for each blot in (A)–(F).
Source: Antonio Galeone et al. Licensed under CC BY 4.0. PMID: 42378087.

Loss of fly Ostγ impairs Dpp/BMP signaling in the embryonic visceral mesoderm and larval midgut Show full caption (A) Schematic of the Ostγ locus and imprecise excision alleles ( Δ4 and Δ5 ). Exons are represented as boxes, with introns depicted as lines. (B) Survival assay showing percentage of eclosed adults from balanced heterozygous progeny ( OstγΔ/+ × OstγΔ/+ ). n ≥ 200 embryos scored per genotype. (C) Third-instar larval midguts from OstγΔ5/+ (control) or OstγΔ5/Δ5 were dissected and fed bromophenol blue (BPB)-supplemented food for 2 h. BPB turns yellow in the acidic region (AZ, acid zone). Note the distinct yellow AZ (dashed box) in control midguts. n = 100. (D) Stage 14 embryos (ventral view) from OstγΔ5/+ and OstγΔ5/Δ5 were fixed and stained with anti-pMad (red). PS3 and PS7 are marked with white arrowheads. (E) Stage 14 embryos from OstγΔ5/+ and OstγΔ5/Δ5 were stained for Labial (red) and Fas3 (green) to mark VM boundaries. White arrowheads mark Labial expression in PS3 and PS7. (F) Western blot of total protein lysates from third-instar larval midguts (genotypes: y w , OstγΔ5/+ , OstγΔ5/Δ5 , and OstγΔ5/Df(2L)Exel7030 ) probed with an anti-Dpp antibody recognizing the prodomain. Tubulin served as a loading control. A representative immunoblot from two independent biological replicates is shown.
Source: Antonio Galeone et al. Licensed under CC BY 4.0. PMID: 42378087.