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BIX

Cat No.:V12839 Purity: ≥98%
BIX (BiP Inducer X) is a novel and potent BiP (Hsp70-5) ER chaperone inducer with anticancer activity.
BIX
BIX Chemical Structure CAS No.: 101714-41-4
Product category: New1
This product is for research use only, not for human use. We do not sell to patients.
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10mg
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Product Description

BIX (BiP Inducer X) is a novel and potent BiP (Hsp70-5) ER chaperone inducer with anticancer activity. It protects neurons from ER stress.

Biological Activity I Assay Protocols (From Reference)
ln Vitro
BiP protein is increased by BiP inducer X (5 μM; 0–12 hours; SK–N–SH cells)[1]. BiP induces the ATF6 pathway, which in turn induces BiP[1]. Cell death caused by ER stress is decreased by BiP inducer X (5 μM; 12 hours; pretreatment with SK-N-SH cells) through suppressed activation of caspases 3/7 and 4[1].
ln Vivo
The use of BiP inducer X lessens the damage caused by cerebral infarction[1]. A 20 μg dose of BiP inducer -34 g (Japan SLC)[1] was administered intracerebroventricularly. The level of BiP protein significantly increased 24 hours following the dose, indicating that BIX treatment produces BiP protein in vivo.
References
[1]. Kudo T, et al. A molecular chaperone inducer protects neurons from ER stress. Cell Death Differ. 2008;15(2):364-375.
[2]. Yoo SA, et al. A novel pathogenic role of the ER chaperone GRP78/BiP in rheumatoid arthritis. J Exp Med. 2012;209(4):871-886.
Additional Infomation
2-(3,4-dihydroxyphenyl)-2-oxoethyl thiocyanate is a member of the class of thiocyanates that is 3,4-dihydroxyacetophenone in which one of the methyl hydrogens has been replaced by the sulfur of a cyanosulfanediyl (-SC#N) group. It has been found to induce the expression of the endoplasmic reticulum chaperone protein GRP78 (78 kDa glucose-regulated protein, BiP, a highly conserved member of the 70 kDa heat shock protein family) leading to an attenuation of the unfolded protein response. It protects neuronal cells and retinal cells from endoplasmic reticulum (ER)-stress induced cell death. It is a member of thiocyanates, an aromatic ketone and a member of catechols.
These protocols are for reference only. InvivoChem does not independently validate these methods.
Physicochemical Properties
Molecular Formula
C9H7NO3S
Molecular Weight
209.22178
Exact Mass
209.014
CAS #
101714-41-4
PubChem CID
16656807
Appearance
Brown to gray solid powder
Density
1.5±0.1 g/cm3
Boiling Point
517.1±45.0 °C at 760 mmHg
Melting Point
127 °C
Flash Point
266.5±28.7 °C
Vapour Pressure
0.0±1.4 mmHg at 25°C
Index of Refraction
1.665
LogP
1.53
Hydrogen Bond Donor Count
2
Hydrogen Bond Acceptor Count
5
Rotatable Bond Count
3
Heavy Atom Count
14
Complexity
260
Defined Atom Stereocenter Count
0
SMILES
N#CSCC(C1C=CC(O)=C(O)C=1)=O
InChi Key
SVFLBLCWKKQKDW-UHFFFAOYSA-N
InChi Code
InChI=1S/C9H7NO3S/c10-5-14-4-9(13)6-1-2-7(11)8(12)3-6/h1-3,11-12H,4H2
Chemical Name
[2-(3,4-dihydroxyphenyl)-2-oxoethyl] thiocyanate
HS Tariff Code
2934.99.9001
Storage

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Shipping Condition
Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)
Solubility Data
Solubility (In Vitro)
DMSO : ~50 mg/mL (~238.98 mM)
Solubility (In Vivo)
Solubility in Formulation 1: ≥ 2.5 mg/mL (11.95 mM) (saturation unknown) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (add these co-solvents sequentially from left to right, and one by one), clear solution.
For example, if 1 mL of working solution is to be prepared, you can add 100 μL of 25.0 mg/mL clear DMSO stock solution to 900 μL of 20% SBE-β-CD physiological saline solution and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C,1 week): Dissolve 2 g SBE-β-CD in 10 mL saline to obtain a clear solution.

 (Please use freshly prepared in vivo formulations for optimal results.)
Preparing Stock Solutions 1 mg 5 mg 10 mg
1 mM 4.7797 mL 23.8983 mL 47.7966 mL
5 mM 0.9559 mL 4.7797 mL 9.5593 mL
10 mM 0.4780 mL 2.3898 mL 4.7797 mL

*Note: Please select an appropriate solvent for the preparation of stock solution based on your experiment needs. For most products, DMSO can be used for preparing stock solutions (e.g. 5 mM, 10 mM, or 20 mM concentration); some products with high aqueous solubility may be dissolved in water directly. Solubility information is available at the above Solubility Data section. Once the stock solution is prepared, aliquot it to routine usage volumes and store at -20°C or -80°C. Avoid repeated freeze and thaw cycles.

Calculator

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What is the mass of compound required to make a 10 mM stock solution in 5 ml of DMSO given that the molecular weight of the compound is 350.26 g/mol?
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What volume of a given 10 mM stock solution is required to make 25 ml of a 25 μM solution?
Using the equation C1V1 = C2V2, where C1=10 mM, C2=25 μM, V2=25 ml and V1 is the unknown:
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g/mol

Molecular Weight Calculator allows you to calculate the molar mass and elemental composition of a compound, as detailed below:

Note: Chemical formula is case sensitive: C12H18N3O4  c12h18n3o4
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Definitions of molecular mass, molecular weight, molar mass and molar weight:
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In vivo Formulation Calculator (Clear solution)
Step 1: Enter information below (Recommended: An additional animal to make allowance for loss during the experiment)
Step 2: Enter in vivo formulation (This is only a calculator, not the exact formulation for a specific product. Please contact us first if there is no in vivo formulation in the solubility section.)
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Calculation results

Working concentration mg/mL;

Method for preparing DMSO stock solution mg drug pre-dissolved in μL DMSO (stock solution concentration mg/mL). Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug.

Method for preparing in vivo formulation:Take μL DMSO stock solution, next add μL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL ddH2O,mix and clarify.

(1) Please be sure that the solution is clear before the addition of next solvent. Dissolution methods like vortex, ultrasound or warming and heat may be used to aid dissolving.
             (2) Be sure to add the solvent(s) in order.

Biological Data
  • Role of GRP78 in synoviocyte survival. (A) Changes in ER sensor proteins in RA synoviocytes treated with 10 µM thapsigargin. The expressions of GRP78, p-eIF2α, ATF-6, BCL-2, and caspase-12 were determined by Western blot analysis. (B) The effect of Grp78 knockdown on synoviocyte survival. 2 d after transfection with siRNA for Grp78, the mRNA and protein expression levels of GRP78 in RA FLSs were determined by RT-PCR and Western blot analysis, respectively (left). The apoptosis of RA FLSs was induced by treating cells with 1 mM SNP for 12 h 2 d after Grp78 siRNA transfection. Degree of cell death was assessed by MTT assay (right). Results are the mean ± SD of more than four independent experiments performed in duplicate. *, P < 0.05 versus control siRNA–transfected cells. (C) RA FLSs were treated with BIX for 12 h, and then GRP78 expression was determined by Western blot analysis. Synoviocyte apoptosis was induced by treating RA FLSs with 10 µM thapsigargin in the presence or absence of BIX. *, P < 0.05 versus thapsigargin-treated cells in the absence of BIX. (D) SV40-immortalized RA FLSs were transfected with either the pFLAG-hGrp78 gene or pFLAG vector only. The protein expression levels for GRP78 were determined by Western blotting. (E and F) RA FLSs transfected with either the pFLAG-hGrp78 gene or pFLAG vector were treated with 5 µM thapsigargin (Tg) for 1 h, 10 µg/ml tunicamycin (Tm) for 12 h, or 5 mM SNP for 24 h. Cell viability was determined by MTT assay. *, P < 0.05 versus vector-transfected cells. (G) The apoptosis of RA FLSs harboring the pFLAG-hGrp78 gene or pFLAG vector was induced by treating cells with 5 µM thapsigargin or 10 µg/ml tunicamycin for 3 h. Degrees of apoptosis were assessed by APOPercentage apoptosis assay, a colorimetric method. Apoptotic cells appeared bright pink. Fold increase in apoptosis levels was expressed as pixel numbers. *, P < 0.05 versus vector-transfected cells. Bars, 100 µm. (C and E–G) Data show mean ± SD.[2]. Yoo SA, et al. A novel pathogenic role of the ER chaperone GRP78/BiP in rheumatoid arthritis. J Exp Med. 2012;209(4):871-886.
  • Increase in synovial hyperplasia and bone erosion in mice treated with BIX, a selective BiP/GRP78 inducer. (A) Hematoxylin and eosin staining of the joints of mice administered periarticularly on alternate days for 2 or 4 wk with BIX. Black arrows and arrowheads in the top panel indicate intact cartilages and minimal synovial proliferation, respectively. Pink arrows in the middle panel indicate the enhanced proliferation of synoviocytes, and black arrows in the bottom panel represent bone erosion. The rectangular area in the middle left image is magnified in the middle right image. (B) The histological scores for degrees of synovial hyperplasia in mice injected with BIX alone, collagenase (COL) alone, and collagenase plus BIX (n = 6 per group). *, P < 0.05 versus collagenase (only)-treated mice without injecting BIX. (C) vWF staining of the synovia of mice treated with collagenase plus BIX versus collagenase alone. Positive cells are shown in brown (black arrows). (D) Evaluation of synovial proliferation by PCNA staining. Positive staining in the synoviums is indicated by brown nuclei. Ratios of positive cells (the number of positive synoviocytes/total synoviocytes counted) are presented in the bottom panel. *, P < 0.001. (B and D) Data show mean ± SD. Bars: (A, top and middle left) 300 µm; (A, middle right and bottom) 120 µm; (C and D) 60 µm.[2]. Yoo SA, et al. A novel pathogenic role of the ER chaperone GRP78/BiP in rheumatoid arthritis. J Exp Med. 2012;209(4):871-886.
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