Recombinant Mouse Stc2 Protein, 25-296aa, C-His tagged

• Cat.No.: STC2-16118M
• Product Overview: Recombinant Mouse Stc2 Protein (25-296aa) with C-His tag was expressed in hek293.

Specification

• Species: Mouse
• Source: HEK293
• Tag: His
• Protein Length: 25-296aa
• Description: Predicted to enable enzyme binding activity; heme binding activity; and protein homodimerization activity. Acts upstream of or within several processes, including cellular calcium ion homeostasis; endoplasmic reticulum unfolded protein response; and regulation of store-operated calcium entry. Located in Golgi apparatus and endoplasmic reticulum. Is expressed in several structures, including embryo mesenchyme; genitourinary system; gut; olfactory epithelium; and vertebral axis musculature. Orthologous to human STC2 (stanniocalcin 2).
• Form: Liquid
• Molecular Mass: Theoretical molecular weight: ~ 30 kDa including His tag.
• AA Sequence: TDSTNPPEGPQDRSSQQKGRLSLQNTAEIQHCLVNAGDVGCGVFECFENNSCEIQGLHGICMTFLHNAGKFDAQGKSFIKDALRCKAHALRHKFGCISRKCPAIREMVFQLQRECYLKHDLCSAAQENVGVIVEMIHFKDLLLHEPYVDLVNLLLTCGEDVKEAVTRSVQAQCEQSWGGLCSILSFCTSNIQRPPTAAPEHQPLADRAQLSRPHHRDTDHHLTANRGAKGERGSKSHPNAHARGRTGGQSAQGPSGSSEWEDEQSEYSDIRRHHHHHH
• Purity: > 90% as determined by SDS-PAGE
• Storage: Short Term Storage at +4 centigrade, Long Term, please prepare aliquots with 20% glycerol and store at -20 to -80 centigrade. Avoid freeze/thaw cycles.
• Concentration: 0.72 mg/mL
• Storage Buffer: PBS, pH 7.4
• Publications:
  Histidine-Rich Glycoprotein and Stanniocalcin-2 High Affinity Interactions with Inflammatory Cells (2022)
  Quartz Crystal Microbalance Measurement of Histidine-Rich Glycoprotein and Stanniocalcin-2 Binding to Each Other and to Inflammatory Cells (2021)
  Leukocyte differentiation by histidine-rich glycoprotein/stanniocalcin-2 complex regulates murine glioma growth through modulation of antitumor immunity (2018)

Gene Information

• Gene Name: Stc2 stanniocalcin 2 [ Mus musculus (house mouse) ]
• Official Symbol: Stc2
• Synonyms: Stc2; stanniocalcin 2; Stc-2; Stc2l; mustc2; stanniocalcin-2
• Gene ID: 20856
• mRNA Refseq: NM_011491
• Protein Refseq: NP_035621
• UniProt ID: O88452

Citation

Quartz Crystal Microbalance Measurement of Histidine-Rich Glycoprotein and Stanniocalcin-2 Binding to Each Other and to Inflammatory Cells

Journal: Cells    Data: 2022/8/29

Authors: Tor Persson Skare, Hiroshi Kaito, Vivek P. Chavda

Article Snippet:The U937 cells were seeded at 10 4 cells per well in 8-well chamber slides (cat. no. 80826, ibidi, Fitchburg, WI, USA).The U937 cells were seeded at 10 4 cells per well in 8-well chamber slides (cat. no. 80826, ibidi, Fitchburg, WI, USA).. At the start of the experiment, cells were incubated with 10 nM vitD3, recombinant, in-house purified HRG (mouse) at 1 μg/mL (13.3 nM) [ ], and STC2 (mouse) (cat. no. STC2-16118 M, Creative BioMart, Shirley, NY, USA) or inactive HRG protein at equivalent molar concentrations together with sterile green E. coli bioparticles (cat. no. 4616, Essen Bioscience, Ann Arbor, MI, USA) at 33 μg/mL.. Following 20 h incubation at 37 °C in 5% CO 2 , cells were imaged at 10× with a Zeiss LSM 700 Microscope with AxioCam HRm and Zen Black software (Zeiss, Oberkochen, Germany).Following 20 h incubation at 37 °C in 5% CO 2 , cells were imaged at 10× with a Zeiss LSM 700 Microscope with AxioCam HRm and Zen Black software (Zeiss, Oberkochen, Germany).

Recombinant HRG binds STC2 and modulates phagocytosis of bioparticles. ( A ) Co-immunoprecipitation of HRG but not inactive HRG with STC2. Recombinant proteins (2 μg each) were separated on SDS-PAGE as individual preparations (loading control) or after mixing and immunoprecipitation (IP) using antibodies against STC2, followed by immunoblotting (IB) as indicated. ( B ) Ratio of HRG (active or inactive) band intensities in the STC2 immunoprecipitate normalized to corresponding active and inactive HRG loading controls. Statistical analysis; Student’s t -test. ( C ) Representative microscope images of U937 monocytes without (left) or with treatment with active HRG (right) in the phagocytosis assay. Green cells have engulfed pH-sensitive fluorescent bioparticles. Scale bar; 50 μm. ( D ) Quantification of phagocytosis efficiency in the different treatment conditions. The proportion of positive (green) phagocytotic U937 cells to all cells per field of vision is shown in relation to the positive cells/total cells in the vitD3 differentiated HRG-treated condition (set to 1). Statistical analysis; Tukey’s multiple comparisons test ( D ). *** p < 0.001; **** p < 0.0001.

Affinity determination of HRG’s binding to STC2 using QCM. ( A ) The immobilization of STC2 on the QCM LNB sensor surface. ( B ) Sensorgram and kinetic analysis showing chip-immobilized STC2 and binding of HRG at three different concentrations: 50, 100 and 200 nM. Black lines: experimental curves. Red lines: fitted curves. For representative sensorgram shown, three injections per concentration, two independent experiments. ( C ) Sensorgram showing chip-immobilized STC2 and lack of binding of inactive HRG tested at three different concentrations: 50, 100 and 200 nM. For representative sensorgram shown, three injections per concentration, two independent experiments. ( D ) Sensorgram showing chip-immobilized HRG and lack of binding of STC2, tested at three different concentrations: 200 nM, 450 nM and 900 nM. For representative sensorgram shown, two injections per concentration, three independent experiments.

Binding of HRG and STC2 individually and together to live U937 cells. ( A ) Schematic outline of the experimental setup. Undifferentiated or vitD3 differentiated U937 cells, immobilized on QCM LNB chips with HRG, STC2 or a mix of the two, injected over chip surfaces. ( B ) Real-time qPCR data of CD14 expression normalized to GAPDH on undifferentiated and vitD3 differentiated U937 cells seeded on the QCM chip. Three independent analyses. ( C – F ) Representative sensorgram showing frequency response from injections over undifferentiated ( C , E , G ) and vitD3 differentiated ( D , F , H ) live U937 cells. Black lines: experimental curves. Red lines: fitted curves of the 1:1 interaction model. For representative sensorgrams shown, three injections per concentration, two independent experiments.

Histidine-Rich Glycoprotein and Stanniocalcin-2 High Affinity Interactions with Inflammatory Cells

Data: 2022/1/17

Authors: Lena Claesson-Welsh, Tor Persson Skare, Teodor Aastrup

Article Snippet:253 254 255 256 10 Purified proteins and phagocytosis assay 257 U937 cells were seeded at 104 cells per well in 8-well chamber slides (ibidi 80826).253 254 255 256 10 Purified proteins and phagocytosis assay 257 U937 cells were seeded at 104 cells per well in 8-well chamber slides (ibidi 80826).. At the start of the 258 experiment, cells were incubated with 10 nM vitD3, recombinant, in-house purified HRG (mouse) at 1 259 μg/ml (13.3 nM) 23, ,strong>STC2 (mouse) (cat no. STC2-16118M, Creative BioMart) or inactive HRG protein at 260 equivalent molar concentrations together with sterile green E. coli bioparticles (cat. no. 4616, Essen 261 Bioscience) at 33 μg/ml.. Following 20h incubation at 37°C in 5% CO2, cells were imaged at 10X with a 262 Zeiss LSM 700 Microscope with AxioCam HRm and Zen software (Zeiss).Following 20h incubation at 37°C in 5% CO2, cells were imaged at 10X with a 262 Zeiss LSM 700 Microscope with AxioCam HRm and Zen software (Zeiss).