Prepare an insulin solution in HEPES buffer by the following sample protocol:
1. Dissolve 10 mg of Insulin in 1 ml of 25 mM HEPES, pH 8.2 – this will result in a cloudy or milky solution.
2. Adjust the pH to less than 3 by adding 5 N HCl, dropwise, until the solution is clear.
3. Titrate back to pH 8.2 using 5 N NaOH, dropwise. The solution may become turbid at pH 7, but will clear above pH 7.5, sterile-filter and store at 2-8°C.
I6634
Insulin from bovine pancreas
powder, BioReagent, suitable for cell culture
Synonym(s):
Bovine insulin, Cell culture grade insulin
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50 MG
$279.00
100 MG
$543.00
250 MG
$959.00
500 MG
$1,770.00
1 G
$2,640.00
About This Item
Empirical Formula (Hill Notation):
C254H377N65O75S6
CAS Number:
Molecular Weight:
5733.49
UNSPSC Code:
12352209
NACRES:
NA.77
EC Number:
234-291-2
MDL number:
Biological source:
bovine
Form:
powder
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InChI key
IXIBAKNTJSCKJM-BUBXBXGNSA-N
biological source
bovine
product line
BioReagent
form
powder
potency
≥25 USP units per mg
technique(s)
cell culture | mammalian: suitable, single cell analysis: suitable
solubility
acetic acid: 2 mg/mL (pH: 2-3), hydrochloric acid: 2 mg/mL (pH: 2-3)
UniProt accession no.
shipped in
ambient
storage temp.
−20°C
Quality Level
Gene Information
cow ... INS(280829)
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1 of 4
This Item | |||
|---|---|---|---|
| biological source bovine | biological source - | biological source bovine | biological source bovine |
| Quality Level 200 | Quality Level 200 | Quality Level 200 | Quality Level 200 |
| product line BioReagent | product line - | product line BioXtra | product line - |
| form powder | form powder | form lyophilized powder | form powder |
| technique(s) cell culture | mammalian: suitable, single cell analysis: suitable | technique(s) - | technique(s) cell culture | mammalian: suitable | technique(s) cell culture | hybridoma: suitable |
| solubility acetic acid: 2 mg/mL (pH: 2-3), hydrochloric acid: 2 mg/mL (pH: 2-3) | solubility water: 2 mg/mL, clear, colorless (pH: 2) | solubility acetic acid: 2 mg/mL (pH: 2-3), hydrochloric acid: 2 mg/mL (pH: 2-3) | solubility acidified water, pH 2.0: 2 mg/mL |
Application
Insulin is used as a growth factor in many mammalian cell culture systems.
- Insulin from bovine pancreas has been used for the differentiation of MSCs (mesenchymal stem cells) to adipocytes.
- It has been used for culturing of various cells.
- It has also been used to study insulin-mediated inhibition of the HMGCS2 (3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) synthase) gene.
Biochem/physiol Actions
Two-chain polypeptide hormone produced by the β-cells of pancreatic islets. Its molecular weight is ~5800 Da. The α and β chains are joined by two interchain disulfide bonds. The α chain contains an intrachain disulfide bond. Insulin regulates the cellular uptake, utilization, and storage of glucose, amino acids, and fatty acids and inhibits the breakdown of glycogen, protein, and fat.
Physical form
powder-0 °C; solution stable 2-8 °C, 1 year
Preparation Note
Insulin has low solubility at neutral pH. It can be solubilized at 2 mg/ml in dilute acetic or hydrochloric acid, pH 2-3.
Other Notes
InChI: 1S/C254H377N65O75S6/c1-29-131(24)205(312-192(334)103-256)250(389)317-204(130(22)23)246(385)287-158(74-81-199(344)345)216(355)281-155(70-77-188(260)330)220(359)307-183-116-399-400-117-184-242(381)305-177(110-321)238(377)293-161(87-122(6)7)223(362)294-167(94-139-52-60-145(324)61-53-139)226(365)282-153(68-75-186(258)328)217(356)289-160(86-121(4)5)221(360)284-157(73-80-198(342)343)219(358)301-173(100-189(261)331)233(372)297-169(96-141-56-64-147(326)65-57-141)229(368)308-182(241(380)303-175(253(393)394)102-191(263)333)115-398-396-113-180(212(351)270-106-193(335)277-152(71-78-196(338)339)215(354)280-150(50-41-83-268-254(264)265)210(349)269-107-194(336)278-165(92-137-45-35-31-36-46-137)225(364)296-166(93-138-47-37-32-38-48-138)228(367)298-170(97-142-58-66-148(327)67-59-142)236(375)318-206(135(28)323)251(390)319-84-42-51-185(319)244(383)285-151(49-39-40-82-255)213(352)276-134(27)252(391)392)310-248(387)202(128(18)19)315-234(373)163(89-124(10)11)291-227(366)168(95-140-54-62-146(325)63-55-140)295-222(361)159(85-120(2)3)288-207(346)132(25)274-214(353)156(72-79-197(340)341)286-245(384)201(127(16)17)314-235(374)164(90-125(12)13)292-231(370)172(99-144-105-267-119-273-144)300-237(376)176(109-320)279-195(337)108-271-211(350)179(112-395-397-114-181(309-243(183)382)240(379)275-133(26)208(347)304-178(111-322)239(378)316-203(129(20)21)249(388)311-184)306-224(363)162(88-123(8)9)290-230(369)171(98-143-104-266-118-272-143)299-218(357)154(69-76-187(259)329)283-232(371)174(101-190(262)332)302-247(386)200(126(14)15)313-209(348)149(257)91-136-43-33-30-34-44-136/h30-38,43-48,52-67,104-105,118-135,149-185,200-206,320-327H,29,39-42,49-51,68-103,106-117,255-257H2,1-28H3,(H2,258,328)(H2,259,329)(H2,260,330)(H2,261,331)(H2,262,332)(H2,263,333)(H,266,272)(H,267,273)(H,269,349)(H,270,351)(H,271,350)(H,274,353)(H,275,379)(H,276,352)(H,277,335)(H,278,336)(H,279,337)(H,280,354)(H,281,355)(H,282,365)(H,283,371)(H,284,360)(H,285,383)(H,286,384)(H,287,385)(H,288,346)(H,289,356)(H,290,369)(H,291,366)(H,292,370)(H,293,377)(H,294,362)(H,295,361)(H,296,364)(H,297,372)(H,298,367)(H,299,357)(H,300,376)(H,301,358)(H,302,386)(H,303,380)(H,304,347)(H,305,381)(H,306,363)(H,307,359)(H,308,368)(H,309,382)(H,310,387)(H,311,388)(H,312,334)(H,313,348)(H,314,374)(H,315,373)(H,316,378)(H,317,389)(H,318,375)(H,338,339)(H,340,341)(H,342,343)(H,344,345)(H,391,392)(H,393,394)(H4,264,265,268)/t131-,132-,133-,134-,135+,149-,150-,151-,152-,153-,154-,155-,156-,157-,158-,159-,160-,161-,162-,163-,164-,165-,166-,167-,168-,169-,170-,171-,172-,173-,174-,175-,176-,177-,178-,179-,180-,181-,182-,183-,184-,185-,200-,201-,202-,203-,204-,205-,206-/m0/s1
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wgk
WGK 3
flash_point_f
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flash_point_c
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ppe
Eyeshields, Gloves, type N95 (US)
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Boah Lee et al.
PloS one, 12(8), e0183569-e0183569 (2017-08-29)
Pancreatic islets can adapt to oscillatory glucose to produce synchronous insulin pulses. Can islets adapt to other oscillatory stimuli, specifically insulin? To answer this question, we stimulated islets with pulses of exogenous insulin and measured their Ca2+ oscillations. We observed
Down-regulation of the mitochondrial 3-hydroxy-3-methylglutaryl-CoA synthase gene by insulin: the role of the forkhead transcription factor FKHRL1.
Nadal A et al.
The Biochemical Journal, 366, 289-289 (2002)
Major histocompatibility complex (MHC) class I gene expression in single neurons of the central nervous system: differential regulation by interferon (IFN)-gamma and tumor necrosis factor (TNF)-alpha.
Neumann H et al.
The Journal of Experimental Medicine, 185, 305-305 (1997)
Lei Zhang et al.
Oncotarget, 7(49), 80382-80390 (2016-11-03)
Pleiotrophin (PTN) augments tumor growth by increasing proliferation of tumor cells and promoting vascular abnormalization, but its role in early gliomagenesis has not been evaluated. Through analysis of publically available datasets, we demonstrate that increased PTN mRNA expression is associated
CCL2/CCR2 chemokine signaling coordinates survival and motility of breast cancer cells through Smad3 protein- and p42/44 mitogen-activated protein kinase (MAPK)-dependent mechanisms.
Fang WB et al.
The Journal of Biological Chemistry, 287, 36593-36593 (2012)
Related Content
Instructions
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How can i make solution of insulin in HEPES buffer and how this solution is storage?
1 answer-
Helpful?
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I'm using this product (I6634) for my culture medium. Recently, I found my GLP-1 ELISA kit detects this insulin powder. I'm wondering if this insulin powder is purified from pancreatic extract using an antibody-based method, or simply a concentrated form.
1 answer-
No antibodies are used in the purification of this product.
Helpful?
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How is your cell culture grade insulin from bovine pancreas product is extracted and purified? Is the pancreas tissue homogenized prior to insulin protein extraction? Are any steps taken to remove contaminants? What is the estimated purity?
1 answer-
The manufacturing method is considered confidential. This product is not tested the purity. The potency is tested for each lot and reported on the Certificate of Analysis.
Please see the link below to access a sample or lot specific Certificate:
https://www.sigmaaldrich.com/product/sigma/i6634#product-documentationHelpful?
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To make a dilution of the stock prepared at 10 mg/ml in HEPES (i-e.10uM which is 34-fold than 2mg/ml) should I still use acidified water, or can i just dilute in PBS or water? This 10 uM dilution will be further dilute 1000 times for cell culture
1 answer-
A stock solution in HEPES at 10 mg/mL, such as that offered as product I0516, may be diluted with water, medium, or the aqueous buffer of choice. This would includes HEPES or PBS. Please see the link below to review the product information sheet:
https://www.sigmaaldrich.com/deepweb/assets/sigmaaldrich/product/documents/132/262/i6634pis.pdfHelpful?
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What is the comparison between International Units (IU) and USP units for insulin?
1 answer-
The international unit (IU) of insulin is based on the International Standard Preparation of Bioassay (1958). The standard preparation is a mixture of 52% bovine and 48% porcine insulin, containing 24 units/mg. Reference: Martindale: The Extra Pharmacopoeia, 29th ed., p. 392. High purity bovine pancreatic insulin typically assays at approximately 27 USP units per mg. So the IU and USP unit are not identical, but they are approximately 1:1.
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How many units of insulin are present in my vial of Product I6634, Insulin from bovine pancreas?
1 answer-
Product No. I6634 has a specification of not less than 27 USP units/mg. The exact value for the product received can be found the on lot-specific Certificate of Analysis.
Helpful?
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What can be used to make a solution of insulin?
1 answer-
Insulin is soluble in dilute acids and alkalis. The solubility of insulin is tested at 2 mg/mL in water (acidified to pH 2 with hydrochloric acid). One can also use 1 mL of water acidified to pH 2 with 10 μL of glacial acetic acid to dissolve 2 mg of insulin. A 10 mg/mL solution of bovine insulin in 25 mM HEPES at pH 8.2 is offered as Product No. I0516.
Helpful?
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How can I store the insulin solution? How long?
1 answer-
The USP recommends that solutions of insulin be stored at 2-8°C. It is not recommended to freeze solutions of insulin, as it can aggregate. Sterile solutions should be stable for at least 1 year at 2-8°C.
Helpful?
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What is the Department of Transportation shipping information for this product?
1 answer-
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
Helpful?
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What is the amino acid sequence for Product I6634, Insulin from bovine pancreas?
1 answer-
The sequence and structure of insulin can be found on the product information sheet (under Documents, above).
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