Plexin B2 Antibody [Unconjugated]

HAF016). A specific band was detected for Plexin B2 at approximately 170 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8." > HAF016). A specific band was detected for Plexin B2 at approximately 170 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8." title="Western blot shows lysates of human brain tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Plexin B2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5329) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Plexin B2 at approximately 170 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8." />

Western blot shows lysates of human brain tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Plexin B2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5329) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # <a class=

CTS013). Tissue was stained with the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific labeling was localized to the plasma membrane and cytoplasm of epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." > CTS013). Tissue was stained with the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific labeling was localized to the plasma membrane and cytoplasm of epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." title="Plexin B2 was detected in immersion fixed paraffin-embedded sections of human colon cancer using 3 µg/mL Sheep Anti-Human Plexin B2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5329) overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained with the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific labeling was localized to the plasma membrane and cytoplasm of epithelial cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections." />

Plexin B2 was detected in immersion fixed paraffin-embedded sections of human colon cancer using 3 µg/mL Sheep Anti-Human Plexin B2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5329) overnight at 4 °C. Before incubation with the primary antibody tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # <a class=

NL010) and counterstained with DAPI (blue). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections." > NL010) and counterstained with DAPI (blue). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections." title="Plexin B2 was detected in perfusion fixed frozen sections of mouse brain (choroid plexus) using 10 µg/mL Sheep Anti-Human Plexin B2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5329) overnight at 4 °C. Tissue was stained with the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections." />

Plexin B2 was detected in perfusion fixed frozen sections of mouse brain (choroid plexus) using 10 µg/mL Sheep Anti-Human Plexin B2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5329) overnight at 4 °C. Tissue was stained with the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # <a class=

HAF016). A specific band was detected for Plexin B2 at approximately 170 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." > HAF016). A specific band was detected for Plexin B2 at approximately 170 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." title="Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Plexin B2 knockout HeLa cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human Plexin B2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5329) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Plexin B2 at approximately 170 kDa (as indicated) in the parental HeLa cell line, but is not detectable in knockout HeLa cell line. GAPDH (Catalog # AF5718) is shown as a loading control. This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1." />

Western blot shows lysates of HeLa human cervical epithelial carcinoma parental cell line and Plexin B2 knockout HeLa cell line (KO). PVDF membrane was probed with 0.5 µg/mL of Sheep Anti-Human Plexin B2 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5329) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # <a class=

Plexin-B2 enhances Rnd3-induced cell rounding and inhibition of invasion. (A,B) HeLa cells were transfected with pCMV5 (empty vector control), FLAG–Rnd3 and/or full-length plexin-B2. After 16–18 h, cells were fixed and stained with anti-FLAG antibody and anti-plexin-B2 antibody. Actin filaments were visualized with AlexaFluor-546- or AlexaFluor-633-conjugated phalloidin. Scale bar: 10 µm. (B) Cell spread area (mean±s.e.m., n=3). Each dot represents a single cell; 19 cells were analysed for each condition. (C) HeLa cells were transfected with GFP (control), or GFP–Rnd3 with or without plexin-B2. Cell invasion was measured using Matrigel-coated Transwell filters. Invasion is shown relative to GFP-expressing control cells (mean±s.e.m., n=35). (D,E) HeLa cells (1×105 cells/ml) were transfected with pCMV5 (control), full-length plexin-B2 and wild-type (WT) FLAG–Rnd3 or FLAG–Rnd3 mutants that do not bind to plexin-B2 (Rnd3-V56Y, Rnd3-V87R and Rnd3-1–200). After 24 h, cells were fixed and stained for plexin-B2 (green) and FLAG–Rnd3 proteins (red). Actin filaments were detected with AlexaFluor-633-conjugated phalloidin (blue). Scale bar: 10 μm. (E) Mean percentage±s.e.m. of rounded cells from three independent experiments; n≥100 cells per experiment. (F) HeLa cells were transfected with two siRNAs targeting plexin-B2 or with control siRNA. After 48 h, cells were transfected with FLAG-tagged Rnd3. Cells were lysed or fixed, permeabilized and immunostained 24 h later. Plexin-B2 depletion was determined by immunoblotting with anti-plexin-B2 antibody. Endogenous Rnd3 and exogenous Rnd3 were detected using anti-Rnd3 or anti-FLAG antibody, respectively. Blots are representative of three independent experiments. (G) Mean percentage±s.e.m. of rounded cells from three independent experiments; n≥100 cells per experiment. For all graphs, *P<0.05, **P<0.01, ***P<0.001 ****P<0.0001; ns, not significant; one-way ANOVA with Tukey posthoc test. Image collected and cropped by CiteAb from th

• Reactivity: Hu
• Applications: WB, IHC, KO
• Clonality: Polyclonal
• Host: Sheep
• Conjugate: Unconjugated
• Concentration: LYOPH

Plexin B2 Antibody [Unconjugated] Summary

• Immunogen: Chinese hamster ovary cell line CHO-derived recombinant human Plexin B2
  Leu20-Arg1160
  Accession # O15031
• Specificity: Detects human Plexin B2 in direct ELISAs and Western blots. In direct ELISAs, approximately 25% cross-reactivity with recombinant mouse Plexin B2 is observed.
• Source: N/A
• Isotype: IgG
• Clonality: Polyclonal
• Host: Sheep
• Gene: PLXNB2
• Purity Statement: Antigen Affinity-purified

Applications/Dilutions

• Dilutions:
  • Immunohistochemistry 5-15 ug/mL
  • Knockout Validated
  • Western Blot 1 ug/mL

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Reconstitute at 0.2 mg/mL in sterile PBS.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Plexin B2 Antibody [Unconjugated]

• MM1
• MM1KIAA0315dJ402G11.3
• Nbla00445
• PLEXB2
• Plexin B2
• plexin-B2
• PLXNB2

Background

Plexin B2 is a 240 kDa member of the B subfamily, Plexin family of semaphorin receptors. It is expressed by cerebellar granule cells, neuroepithelium and chondrocytes and serves as a receptor for Sema4C. Mature human Plexin B2 is 1819 amino acid (aa) in length. It is a type I transmembrane (TM) glycoprotein that contains an 1178 aa extracellular domain (ECD) plus a 620 aa cytoplasmic region. The ECD contains one semaphorin domain (aa 20‑466) and three IPT repeats (aa 803‑1092). The ECD may be cleaved into two subunits, a 170 kDa alpha ‑chain (aa 20‑1164) and an 80 kDa TM beta -chain that remain noncovalently-linked. Multiple splice variants may exist. One shows an alternate start site at Met1369, while a second shows a deletion of aa 67‑638. Over aa 20‑1160, human Plexin B2 is 82% aa identical to mouse Plexin B2.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Plexin B2 Antibody (AF5329)(9)

Publications using AF5329

• Alves, CJ;Hannah, T;Sadia, S;Kolsteeg, C;Dixon, A;Wiener, RJ;Nguyen, H;Tipping, MJ;Ladeira, JS;Franklin, PFDC;Dutra de Nigro, NP;Dias, RA;Zabala Capriles, PV;Rodrigues Furtado de Mendonça, JP;Slesinger, P;Costa, K;Zou, H;Friedel, RH; Invasion of glioma cells through confined space requires membrane tension regulation and mechano-electrical coupling via Plexin-B2 bioRxiv : the preprint server for biology 2024-01-02 [PMID: 38313256] (Western Blot, ICC, Human)
• Anju Karki, Reshma Purohit, Sofia Tosoni, Narendra Bharathy, Joel E Michalek, Sonja Chen et al. Plexin-B2 and Semaphorins Do Not Drive Rhabdomyosarcoma Proliferation or Migration Sarcoma 5/6/2022 [PMID: 35570846]
• C Junqueira, R Dariolli, J Haydak, S Kang, T Hannah, RJ Wiener, S DeFronzo, R Tejero, GL Gusella, A Ramakrishn, R Alves Dias, A Wojcinski, S Kesari, L Shen, EA Sobie, JP Rodrigues, EU Azeloglu, H Zou, RH Friedel Plexin-B2 orchestrates collective stem cell dynamics via actomyosin contractility, cytoskeletal tension and adhesion Nature Communications, 2021-10-14;12(1):6019. 2021-10-14 [PMID: 34650052] (ICC, Western Blot, Human)
• Chen Jiang, Ahsan Javed, Laura Kaiser, Michele M. Nava, Rui Xu, Dominique T. Brandt et al. Mechanochemical control of epidermal stem cell divisions by B-plexins Nature Communications 2/26/2021 [PMID: 33637728]
• Chen Jiang, Ahsan Javed, Laura Kaiser, Michele M. Nava, Rui Xu, Dominique T. Brandt et al. Mechanochemical control of epidermal stem cell divisions by B-plexins Nature Communications 2021-02-26 [PMID: 33637728] (Human)
• Yong Huang, Rut Tejero, Vivian K. Lee, Concetta Brusco, Theodore Hannah, Taylor B. Bertucci et al. Plexin-B2 facilitates glioblastoma infiltration by modulating cell biomechanics Communications Biology 1/29/2021 [PMID: 33514835]
• Yong Huang, Rut Tejero, Vivian K. Lee, Concetta Brusco, Theodore Hannah, Taylor B. Bertucci et al. Plexin-B2 facilitates glioblastoma infiltration by modulating cell biomechanics Communications Biology 2021-01-29 [PMID: 33514835] (Western Blot, Mouse)
• Iratxe Zuazo-Gaztelu, Marta Pàez-Ribes, Patricia Carrasco, Laura Martín, Adriana Soler, Mar Martínez-Lozano et al. Anti-tumor effects of anti-Semaphorin 4D antibody unravel a novel pro-invasive mechanism of vascular targeting agents Cancer Research 10/15/2019 [PMID: 31239269]
• Rnd3-induced cell rounding requires interaction with Plexin-B2 J Cell Sci, 2016-09-21;0(0):. 2016-09-21 [PMID: 27656111] (IHC, Human)

Bioinformatics

• Gene Symbol: PLXNB2
• Uniprot:
  • O15031()