Mouse Spi-B Antibody

Catalog # Availability Size / Price Qty
AF7204
AF7204-SP
Spi-B in Mouse Splenocytes.
5 Images
Product Details
Citations (7)
FAQs
Supplemental Products
Reviews

Mouse Spi-B Antibody Summary

Species Reactivity
Mouse
Specificity
Detects mouse Spi-B in direct ELISAs. In direct ELISAs, less than 2% cross-reactivity with recombinant human Spi-B is observed.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant mouse Spi-B
Tyr18-Glu167 (Tyr110Phe)
Accession # O35906
Formulation
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.

Applications

Recommended Concentration
Sample
CyTOF-ready
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
 
Immunocytochemistry
5-15 µg/mL
See below
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
See below

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

Scientific Data

Immunocytochemistry Spi-B antibody in Mouse Splenocytes by Immunocytochemistry (ICC). View Larger

Spi-B in Mouse Splenocytes. Spi-B was detected in immersion fixed mouse splenocytes using Sheep Anti-Mouse Spi-B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7204) at 15 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). Specific staining was localized to plasma membranes and cytoplasm. View our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Intracellular Staining by Flow Cytometry Detection of SPi-B antibody in Mouse Splenocytes antibody by Flow Cytometry. View Larger

Detection of SPi-B in Mouse Splenocytes by Flow Cytometry. Mouse splenocytes were stained with Sheep Anti-Mouse Spi-B Antigen Affinity-purified Polyclonal Antibody (Catalog # AF7204) followed by Allophycocyanin-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # F0127) and Rat Anti-Mouse B220/CD45R PE-conjugated Monoclonal Antibody (Catalog # FAB1217P). Quadrant markers were set based on control antibody staining (Catalog # 5-001-A). To facilitate intracellular staining, cells were fixed with paraformadehyde and permeabilized with saponin.

Immunocytochemistry/ Immunofluorescence Detection of Mouse Spi-B by Immunocytochemistry/Immunofluorescence View Larger

Detection of Mouse Spi-B by Immunocytochemistry/Immunofluorescence OPGhigh M cells cluster more in cecal patches than in Peyer’s patches.a Whole-mount immunostaining of the FAE of Peyer’s patches (left) and cecal patches (right) for OPG (green) and Spi-B (red). Nuclei were stained with DAPI (blue); scale bars: 50 µm. b Scatter plots of the fluorescence intensities of OPG versus Spi-B. Red dots represent cells stained with anti-Spi-B antibody that was conjugated with HyLyte Fluor 555 and anti-OPG antibody that was conjugated with HyLyte Fluor 647. Blue dots represent background fluorescence intensity of randomly selected non-stained cells. Fluorescence intensities were measured for at least 3000 cells from five FAEs of three mice. c Frequencies of OPGhigh M cells in Peyer’s patches and cecal patches were quantified. *p < 0.05, ***p < 0.005. Student’s t-test, n = 5 FAE from three animals. The source data underlying panels b and c are provided as a Source Data file. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31932605), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunocytochemistry/ Immunofluorescence Detection of Mouse Spi-B by Immunocytochemistry/Immunofluorescence View Larger

Detection of Mouse Spi-B by Immunocytochemistry/Immunofluorescence RANKL–RANK signaling is stimulated in the gut epithelia of Opg−/− mice.a GP2+ cells (red) are more effectively induced in the cecal epithelium (CE) and ileal villi (VE) of Opg−/− mice by RANKL administration. Whole-mount immunohistochemical images of Spi-B (green) and GP2 (red) in the VE and CE of WT (upper panels) and Opg−/− mice (lower panels) treated with either GST (control; left) or GST-RANKL (right). Scale bars: 100 µm. Representative images from three independent experiment are shown. b Quantitative PCR analysis of M-cell marker expression in conventional epithelia from the VE and CE of mice injected with GST (control) or GST-RANKL. Results were normalized to Gapdh expression and are presented relative to the expression in the ileal epithelium from GST-treated mice. Data shown are mean values from three independent experiments (error bars indicate standard deviation). ***p < 0.005, **p < 0.01, *p < 0.05; p values were calculated with the Student’s t-test (n = 3 biologically independent experiments). c, d Nuclear translocation activities of RelB and p52 following RANKL stimulation are enhanced in Opg−/− mice. c Western blot analysis of p100/p52 and RelB in the VE and CE of mice injected with GST or GST-RANKL. Rpt4, a subunit of the 26S proteasome, was used as an internal control for the cytoplasmic fraction. Lamin A/C (Lamin) was used as an internal control for the nuclear fraction. Data are representative of two independent experiments. d Right, single confocal planes of the cecal FAE from WT and Opg−/− mice. FAE monolayers were stained with anti-RelB (green) and anti-Spi-B (red) antibodies and with Hoechst 33342. Left, a bar graph summarizing the proportions of RelB-positive cells among total numbers of M cells (Spi-B-positive cells). ***p < 0.005; p values were calculated with the Student’s t-test (n = 4 biologically independent experiments). Scale bars: 20 µm. The source data underlying panels b and d and non-cropped scan images of western blotting (c) are provided as a Source Data file. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31932605), licensed under a CC-BY license. Not internally tested by R&D Systems.

Immunocytochemistry/ Immunofluorescence Detection of Mouse Spi-B by Immunocytochemistry/Immunofluorescence View Larger

Detection of Mouse Spi-B by Immunocytochemistry/Immunofluorescence RANKL–RANK signaling is stimulated in the gut epithelia of Opg−/− mice.a GP2+ cells (red) are more effectively induced in the cecal epithelium (CE) and ileal villi (VE) of Opg−/− mice by RANKL administration. Whole-mount immunohistochemical images of Spi-B (green) and GP2 (red) in the VE and CE of WT (upper panels) and Opg−/− mice (lower panels) treated with either GST (control; left) or GST-RANKL (right). Scale bars: 100 µm. Representative images from three independent experiment are shown. b Quantitative PCR analysis of M-cell marker expression in conventional epithelia from the VE and CE of mice injected with GST (control) or GST-RANKL. Results were normalized to Gapdh expression and are presented relative to the expression in the ileal epithelium from GST-treated mice. Data shown are mean values from three independent experiments (error bars indicate standard deviation). ***p < 0.005, **p < 0.01, *p < 0.05; p values were calculated with the Student’s t-test (n = 3 biologically independent experiments). c, d Nuclear translocation activities of RelB and p52 following RANKL stimulation are enhanced in Opg−/− mice. c Western blot analysis of p100/p52 and RelB in the VE and CE of mice injected with GST or GST-RANKL. Rpt4, a subunit of the 26S proteasome, was used as an internal control for the cytoplasmic fraction. Lamin A/C (Lamin) was used as an internal control for the nuclear fraction. Data are representative of two independent experiments. d Right, single confocal planes of the cecal FAE from WT and Opg−/− mice. FAE monolayers were stained with anti-RelB (green) and anti-Spi-B (red) antibodies and with Hoechst 33342. Left, a bar graph summarizing the proportions of RelB-positive cells among total numbers of M cells (Spi-B-positive cells). ***p < 0.005; p values were calculated with the Student’s t-test (n = 4 biologically independent experiments). Scale bars: 20 µm. The source data underlying panels b and d and non-cropped scan images of western blotting (c) are provided as a Source Data file. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31932605), licensed under a CC-BY license. Not internally tested by R&D Systems.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

=
÷

Preparation and Storage

Reconstitution
Sterile PBS to a final concentration of 0.2 mg/mL.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & 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.

Background: Spi-B

Spi-B (Transcription factor Spi-B) is a 33-45 kDa member of the ets family of transcription factors. It is found in hematopoietic cells such as B cells and plasmacytoid dendritic cells (DC). In transitional B cells, Spi-B promotes their differentiation into follicular (naïve) B cells. In hematopoietic stem cells, Spi-B stimulates the generation of IFN-producing plasmacytoid DC at the expense of T, B and NK cell development. Mouse Spi-B is 267 amino acids (aa) in length. It contains a dual transactivation region (aa 1-62), plus a PEST domain (aa 110-170) and an Ets DNA-binding domain (aa 174-257). There are two isoform variants. One shows a nine aa substitution for aa 1-8, while a second possesses an 18 aa insertion after Leu17. Over aa 18-167, mouse Spi-B shares 91% and 74% aa identity with rat and human Spi-B, respectively.

Long Name
Transcription Factor Spi [Spleen FFV Proviral Integration Oncogene] B
Entrez Gene IDs
6689 (Human); 272382 (Mouse); 499146 (Rat)
Alternate Names
Spi-B transcription factor (Spi-1/PU.1 related); SpiB; Spi-B; transcription factor Spi-B

Product Datasheets

You must select a language.

x

Citations for Mouse Spi-B Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

7 Citations: Showing 1 - 7
Filter your results:

Filter by:

  1. ONECUT2 regulates RANKL-dependent enterocyte and microfold cell differentiation in the small intestine; a multi-omics study
    Authors: MV Luna Velez, HK Neikes, RR Snabel, Y Quint, C Qian, A Martens, GJC Veenstra, MR Freeman, SJ van Heerin, M Vermeulen
    Nucleic Acids Research, 2023-02-22;0(0):.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  2. The gut microbiota induces Peyer's-patch-dependent secretion of maternal IgA into milk
    Authors: K Usami, K Niimi, A Matsuo, Y Suyama, Y Sakai, S Sato, K Fujihashi, H Kiyono, S Uchino, M Furukawa, J Islam, K Ito, T Moriya, Y Kusumoto, M Tomura, RC Hovey, J Sugawara, H Yoneyama, H Kitazawa, K Watanabe, H Aso, T Nochi
    Cell Reports, 2021-09-07;36(10):109655.
    Species: Transgenic Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  3. Osteoprotegerin-dependent M cell self-regulation balances gut infection and immunity
    Authors: S Kimura, Y Nakamura, N Kobayashi, K Shiroguchi, E Kawakami, M Mutoh, H Takahashi-, T Yamada, M Hisamoto, M Nakamura, N Udagawa, S Sato, T Kaisho, T Iwanaga, K Hase
    Nat Commun, 2020-01-13;11(1):234.
    Species: Mouse
    Sample Types: Whole Cells
    Applications: ICC/IF
  4. The role of CSF1R-dependent macrophages in control of the intestinal stem-cell niche
    Authors: A Sehgal, DS Donaldson, C Pridans, KA Sauter, DA Hume, NA Mabbott
    Nat Commun, 2018-03-28;9(1):1272.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC
  5. Development of intestinal M cells and follicle-associated epithelium is regulated by TRAF6-mediated NF-?B signaling
    Authors: T Kanaya, S Sakakibara, T Jinnohara, M Hachisuka, N Tachibana, S Hidano, T Kobayashi, S Kimura, T Iwanaga, T Nakagawa, T Katsuno, N Kato, T Akiyama, T Sato, IR Williams, H Ohno
    J. Exp. Med., 2018-01-16;0(0):.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC-P
  6. Increased Abundance of M Cells in the Gut Epithelium Dramatically Enhances Oral Prion Disease Susceptibility
    PLoS Pathog, 2016-12-14;12(12):e1006075.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC-Fr
  7. c-Rel is dispensable for the differentiation and functional maturation of M cells in the follicle-associated epithelium
    Immunobiology, 2016-09-18;0(0):.
    Species: Mouse
    Sample Types: Whole Tissue
    Applications: IHC

FAQs

No product specific FAQs exist for this product, however you may

View all Antibody FAQs

Reviews for Mouse Spi-B Antibody

There are currently no reviews for this product. Be the first to review Mouse Spi-B Antibody and earn rewards!

Have you used Mouse Spi-B Antibody?

Submit a review and receive an Amazon gift card.

$25/€18/£15/$25CAN/¥75 Yuan/¥1250 Yen for a review with an image

$10/€7/£6/$10 CAD/¥70 Yuan/¥1110 Yen for a review without an image

Submit a Review