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B 1235

Manufactured by Vector Laboratories
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Sourced in United States

The B-1235 is a laboratory instrument designed for the detection and quantification of various biological molecules. It utilizes an array of sensors to analyze samples and provide accurate measurements. The core function of the B-1235 is to facilitate precise analyte detection, offering researchers a valuable tool for their scientific investigations.

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Lab products found in correlation

Chemiluminescence reagent, by Thermo Fisher Scientific (2 mentions) Vicia villosa, by Vector Laboratories (2 mentions) Sambucus niagra, by Vector Laboratories (2 mentions) B 1305, by Vector Laboratories (2 mentions) Dynabeads protein g, by Thermo Fisher Scientific (2 mentions) Complete protease inhibitor, by Roche (2 mentions) Ecl kit, by Thermo Fisher Scientific (2 mentions) Ab178876, by Abcam (1 mentions) Affinipure goat anti mouse igg, by Proteintech (1 mentions) Anti n cadherin, by Santa Cruz Biotechnology (1 mentions) Anti e cadherin, by Thermo Fisher Scientific (1 mentions) Sa00001 1, by Proteintech (1 mentions) Streptavidin hrp, by Thermo Fisher Scientific (1 mentions) Ab115957, by Abcam (1 mentions) Streptavidin hrp, by Vector Laboratories (1 mentions) Ab166898, by Abcam (1 mentions) B 1305 2, by Vector Laboratories (1 mentions) Ab21593, by Abcam (1 mentions) Rneasy plus tissue mini kit, by Qiagen (1 mentions) Fl 1231, by Vector Laboratories (1 mentions)

6 protocols using b 1235

1

Immunoblotting Analysis of EMT Markers

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Cells in experimental and control groups were lyzed and centrifuged. After determined the protein concentrations, the cell lysate samples (30 μg/lane) were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) on 10% gels and transferred onto NC membranes. The membranes were blocked with 5% BSA in TBST and immunoblotted by the primary antibodies. The primary antibodies used in this study were:anti-GALNT6 (1:1000, sc-100755, Santa Cruz Biotechnology), anti-E-cadherin (1:500, 13-1700, Invitrogen), anti-N-cadherin (1:200, sc-393933, Santa Cruz Biotechnology), anti-Slug (1:500, ab27568, Abcam), anti-GRP78 (1:1000, ab21685, Abcam), anti-phospho-MEK1/2 (1:1000, #9154, CST), anti-MEK1/2 (1:1000, ab178876, Abcam), anti-phospho-ERK1/2 (1:1000, #4370, CST), anti-ERK1/2 (1:1000, #4695, CST), biotinylated anti-Viciavillosa agglutinin (specific to GalNAc-Ser/Thr) (VVA, 1:1000, B-1235, Vector) and anti-GAPDH (1:2000, 10494-1-AP, Proteintech). The secondary antibodies were HRP-conjugated Affinipure Goat Anti-Mouse IgG (1:5000, SA00001-1, Proteintech), HRP-conjugated Affinipure Goat Anti-Rabbit IgG (1:5000, SA00001-2, Proteintech) and HRP-streptavidin (1:10000, SA-5014, Vector). The protein bands were visualized by enhanced chemiluminescent reagents (Advansta) and the intensity of protein band was quantified by ImageJ software.
Song J., Liu W., Wang J., Hao J., Wang Y., You X., Du X., Zhou Y., Ben J., Zhang X., Ye M, & Wang Q. (2020). GALNT6 promotes invasion and metastasis of human lung adenocarcinoma cells through O-glycosylating chaperone protein GRP78. Cell Death & Disease, 11(5), 352.
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2

Quantification of Glycosylated P-gp

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Total protein was extracted from indicated cells. P-gp was immunoprecipitated from the cell lysates and subjected to Vicia Villosa Lectin (VVL) blot. After separation by 10% SDSPAGE, samples were transferred to NC membrane that was blocked in blocking solution (5% BSA in TBST). Membrane was incubated with a 0.2 μg/ml biotinylated VVL solution (B-1235, VECTOR laboratories) and then incubated with Streptavidin-HRP (S911, Thermo Scientific). The result was visualized by an ECL kit (Thermo Scientific).
Shan J., Liu Y., Wang Y., Li Y., Yu X, & Wu C. (2018). GALNT14 Involves the Regulation of Multidrug Resistance in Breast Cancer Cells. Translational Oncology, 11(3), 786-793.
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3

Immunoprecipitation and Glycan Analysis of CD44

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Cell lysates were harvested in chilled RIPA buffer containing complete protease inhibitor (Roche). After centrifugation of 3.0 × 104 RPM for 20 minutes at 4°C, supernatants were collected, protein concentration was measured, and equal amounts of lysates were used for immunoprecipitation per manufactures instructions. Briefly, immunoprecipitation was performed with Dynabeads Protein G (Thermo, 10004D) against CD44 antibody (CST) for 1hr hour at 4°C with shaking and rotation. After that, precipitants were added to magnetic bead-antibody complexes for 30-minute incubation at room temperature under shaking and rotation. The immunocomplexes were washed with PBST three times and eluted with sample buffer for 5 minutes at 70°C. The immunoprecipitated proteins were then separated by SDS-PAGE and transferred to polyvinylidene difluoride membrane (PVDF). For the analysis of glycans by lectin blot analysis, 3% BSA was used for blocking, Vicia Villosa (Vector Labs, B-1235) or Sambucus Niagra (Vector Labs, B-1305) biotinylated lectin overnight at 4°C, and streptavidin-horse radish peroxidase (HRP) was applied to visualize bands using chemiluminescence reagents (Thermo). For immunoblot analysis, specific antibodies and secondary HRP-conjugated anti-mouse or anti-rabbit antibodies were used. Detailed antibody information is listed in Supplementary Table 1.
Leon F., Seshacharyulu P., Nimmakalaya R.K., Chugh S., Karmakar S., Nallasamy P., Vengoji R., Rachagani S., Cox J.L., Mallya K., Batra S.K, & Ponnusamy M.P. (2021). Reduction in O-glycome induces differentially glycosylated CD44 to promote stemness and metastasis in pancreatic cancer. Oncogene, 41(1), 57-71.
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4

Immunoprecipitation and Glycan Analysis of CD44

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Cell lysates were harvested in chilled RIPA buffer containing complete protease inhibitor (Roche). After centrifugation of 3.0 × 104 RPM for 20 minutes at 4°C, supernatants were collected, protein concentration was measured, and equal amounts of lysates were used for immunoprecipitation per manufactures instructions. Briefly, immunoprecipitation was performed with Dynabeads Protein G (Thermo, 10004D) against CD44 antibody (CST) for 1hr hour at 4°C with shaking and rotation. After that, precipitants were added to magnetic bead-antibody complexes for 30-minute incubation at room temperature under shaking and rotation. The immunocomplexes were washed with PBST three times and eluted with sample buffer for 5 minutes at 70°C. The immunoprecipitated proteins were then separated by SDS-PAGE and transferred to polyvinylidene difluoride membrane (PVDF). For the analysis of glycans by lectin blot analysis, 3% BSA was used for blocking, Vicia Villosa (Vector Labs, B-1235) or Sambucus Niagra (Vector Labs, B-1305) biotinylated lectin overnight at 4°C, and streptavidin-horse radish peroxidase (HRP) was applied to visualize bands using chemiluminescence reagents (Thermo). For immunoblot analysis, specific antibodies and secondary HRP-conjugated anti-mouse or anti-rabbit antibodies were used. Detailed antibody information is listed in Supplementary Table 1.
Leon F., Seshacharyulu P., Nimmakalaya R.K., Chugh S., Karmakar S., Nallasamy P., Vengoji R., Rachagani S., Cox J.L., Mallya K., Batra S.K, & Ponnusamy M.P. (2021). Reduction in O-glycome induces differentially glycosylated CD44 to promote stemness and metastasis in pancreatic cancer. Oncogene, 41(1), 57-71.
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5

Glycoprotein Analysis in Pancreatic Tissue

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These experiments were performed as we previously described21 (link)–23 (link). For Western blot, antibodies for GAPDH (sc-32233; Santa Cruz Biotechnology, Santa Cruz, CA, USA), Cosmc (sc-67480; Santa Cruz Biotechnology), CEL (ab79131; Abcam, Cambridge, UK; sc-34883; Santa Cruz Biotechnology), DMBT1 (MAB59151; R&D Systems, Minneapolis, MN, USA), elastase (ab21593; Abcam) and trypsin (ab166898; Abcam) were used. Ten micrograms per milliliter of Vicia villosa lectin (VVA) reactive against O-GalNAc (Tn antigen) (B-1235; Vector Laboratories, Burlingame, CA, USA) complexed with 1 µg streptavidin-HRP (21126; Pierce, Thermo Fisher Scientific, Grand Island, NY, USA) was used. For immunohistochemistry, antibodies for Tn Antigen (MA180055; Thermo Fisher Scientific, Grand Island, NY, USA), STn Antigen (ab115957; Abcam), insulin (8138; CST, Beverly, MA, USA) and VVA-fluorescein (FL-1231; Vector Laboratories) were used at a dilution of 1:10023 (link). Ten micrograms per milliliter of Sambucus nigra lectin (SNA) (B- 1305–2; Vector Laboratories, Burlingame, CA, USA) was complexed with 1 µg streptavidin-HRP. For real-time PCR, RNA was extracted with the RNeasy Plus Tissue Mini Kit (Qiagen).
Wolters-Eisfeld G., Mercanoglu B., Hofmann B.T., Wolpers T., Schnabel C., Harder S., Steffen P., Bachmann K., Steglich B., Schrader J., Gagliani N., Schlüter H., Güngör C., Izbicki J.R., Wagener C, & Bockhorn M. (2018). Loss of complex O-glycosylation impairs exocrine pancreatic function and induces MODY8-like diabetes in mice. Experimental & Molecular Medicine, 50(10), 133.
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6

Glycosylation Analysis of EFEMP-2 in Breast Cancer

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Total protein was extracted from MCF-7 or MBA-MD-231 cells or MCF-7 or MBA-MD-231 cells transfected with shRNA targeting GALNT14 (shRNA-T14) or control shRNA (shNC). EFEMP-2 protein also was immunoprecipitated from MCF-7 or MBA-MD-231 cells and subjected to Vicia Villosa Lectin (VVL) blot. After being separated by 10% SDS-PAGE, proteins were transferred to NC membrane, and NC membrane was blocked in blocking solution (5% BSA in TBST). Membrane was incubated with a 0.2-μg/mL biotinylated VVL solution (B-1235, VECTOR Laboratories) and then membrane was incubated in nsecondary antibody: Streptavidin HRP conjugate (S911, Thermo Scientific). The result was visualized by an enhanced chemiluminescence kit (Thermo Scientific).
Zuo T., Shan J., Liu Y., Xie R., Yu X, & Wu C. (2018). EFEMP2 Mediates GALNT14-Dependent Breast Cancer Cell Invasion. Translational Oncology, 11(2), 346-352.
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