11376497001

Roche

Insulin, human

recombinant (yeast)

Synonym(s):
Insulin human
Empirical Formula (Hill Notation):
C257H383N65O77S6
CAS Number:
Molecular Weight:
5807.57
MDL number:

Quality Level

sterility

non-sterile; 0.2 μm filtered

assay

>98% (SDS-PAGE)

form

lyophilized (clear, colorless solution after reconstitution)

specific activity

>26 units/mg protein (At least the same specific activity (EC<sub>50</sub>) compared to the indicated standard is guaranteed.)

mol wt

5,700 Da

packaging

pkg of 100 mg

manufacturer/tradename

Roche

application(s)

cell culture | mammalian: suitable

impurities

<0.1 EU/μg endotoxin (LAL test)

solubility

water: soluble

UniProt accession no.

shipped in

wet ice

storage temp.

2-8°C

InChI key

PBGKTOXHQIOBKM-FHFVDXKLSA-N

Gene Information

human ... INS(3630)

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General description

Recombinant Insulin, human, is produced in yeast and purified by standard chromatographic techniques. Insulin regulates blood glucose. In muscles and adipocytes it stimulates glucose intake and metabolism. It also controls the expression and various enzymes.

Specificity

Insulin, human, is active on most mammalian cells.

Application

Insulin shows a broad range of activities on a variety of somatic cells. Recombinant human insulin can be used to stimulate growth and proliferation of cultured cells and to investigate insulin activity on sensitive cells used in research studies. It is also a component of serum-free media formulations for most primary cells and cell lines.

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.

Other Notes

InChI: 1S/C257H383N65O77S6/c1-29-131(23)205(313-193(339)104-259)252(393)317-204(130(21)22)248(389)288-159(75-82-200(349)350)217(358)282-156(71-78-189(263)335)221(362)308-183-116-403-404-117-184-243(384)305-178(111-324)240(381)294-162(88-123(7)8)225(366)295-168(95-140-53-61-146(329)62-54-140)228(369)283-154(69-76-187(261)333)218(359)290-161(87-122(5)6)223(364)285-158(74-81-199(347)348)220(361)302-174(101-190(264)336)235(376)298-170(97-142-57-65-148(331)66-58-142)231(372)309-182(242(383)304-176(255(396)397)103-192(266)338)115-402-401-114-181(214(355)273-107-194(340)278-153(72-79-197(343)344)216(357)281-151(51-42-84-271-257(267)268)212(353)272-108-195(341)279-166(93-138-46-36-32-37-47-138)227(368)297-167(94-139-48-38-33-39-49-139)230(371)299-171(98-143-59-67-149(332)68-60-143)238(379)320-208(135(27)327)254(395)322-85-43-52-186(322)246(387)286-152(50-40-41-83-258)222(363)321-209(136(28)328)256(398)399)311-250(391)203(129(19)20)316-236(377)164(90-125(11)12)292-229(370)169(96-141-55-63-147(330)64-56-141)296-224(365)160(86-121(3)4)289-210(351)133(25)277-215(356)157(73-80-198(345)346)287-247(388)202(128(17)18)315-237(378)165(91-126(13)14)293-233(374)173(100-145-106-270-120-276-145)301-239(380)177(110-323)280-196(342)109-274-213(354)180(113-400-405-118-185(310-244(183)385)245(386)319-207(134(26)326)253(394)306-179(112-325)241(382)318-206(132(24)30-2)251(392)312-184)307-226(367)163(89-124(9)10)291-232(373)172(99-144-105-269-119-275-144)300-219(360)155(70-77-188(262)334)284-234(375)175(102-191(265)337)303-249(390)201(127(15)16)314-211(352)150(260)92-137-44-34-31-35-45-137/h31-39,44-49,53-68,105-106,119-136,150-186,201-209,323-332H,29-30,40-43,50-52,69-104,107-118,258-260H2,1-28H3,(H2,261,333)(H2,262,334)(H2,263,335)(H2,264,336)(H2,265,337)(H2,266,338)(H,269,275)(H,270,276)(H,272,353)(H,273,355)(H,274,354)(H,277,356)(H,278,340)(H,279,341)(H,280,342)(H,281,357)(H,282,358)(H,283,369)(H,284,375)(H,285,364)(H,286,387)(H,287,388)(H,288,389)(H,289,351)(H,290,359)(H,291,373)(H,292,370)(H,293,374)(H,294,381)(H,295,366)(H,296,365)(H,297,368)(H,298,376)(H,299,371)(H,300,360)(H,301,380)(H,302,361)(H,303,390)(H,304,383)(H,305,384)(H,306,394)(H,307,367)(H,308,362)(H,309,372)(H,310,385)(H,311,391)(H,312,392)(H,313,339)(H,314,352)(H,315,378)(H,316,377)(H,317,393)(H,318,382)(H,319,386)(H,320,379)(H,321,363)(H,343,344)(H,345,346)(H,347,348)(H,349,350)(H,396,397)(H,398,399)(H4,267,268,271)/t131-,132-,133-,134+,135+,136+,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-,186-,201-,202-,203-,204-,205-,206-,207-,208-,209-/m0/s1
For life science research only. Not for use in diagnostic procedures.

Quality

Endotoxin level: <0.1EU/μg (LAL-test)
Note: 1 EU corresponds to 0.1ng

Sequence

A-chain: 21 AS, B-chain: 30 AS, connected by two disulfide bridges
Two polypeptide chains (A-chain: 21 amino acids, B-chain: 30 amino acids, connected by two disulfide bridges) identical to natural, human insulin.

Unit Definition

EC50 definition: The concentration of human insulin that is required to support half-maximal stimulation of cell proliferation (MTT cleavage) with 3T3 (A31) cells.

Physical form

Lyophilizate from a hydrochloric acid solution (pH 2.3) (crystalline insulin)

Preparation Note

Working concentration: 1-10 μg/ml
Recommended concentration for serum-free cell culture is 1-10 μg/ml.
The concentration of insulin required for stimulation of cell growth in almost all cases is extraordinarily high compared with the physiological concentration. Insulin may be mimicking insulin-like growth factors (IGFs, somatomedins) for some cell lines, and high insulin concentrations may be necessary to occupy receptors which have a high affinity for IGFs and a lower affinity for insulin.
Working solution: Dissolve Insulin, human, recombinant (100 mg or 500 mg), in sterile, double-dist. water (final concentration: 10 mg/ml).
Storage conditions (working solution): -15 to -25 °C
The reconstituted, undiluted solution is stable at -15 to -25 °C.

Note: Avoid repeated freezing and thawing.

Reconstitution

Reconstitute in sterile double-distilled water (final concentration 10 mg/ml), further dilution with PBS (phosphate buffered saline) or medium containing 1 mg/ml (0.1%) BSA (bovine serum albumin) [or HSA (human serum albumin)], or 1 to 10% serum.

storage_class_code

11 - Combustible Solids

WGK Germany

nwg

Flash Point(F)

Not applicable

Flash Point(C)

Not applicable

Certificate of Analysis

Certificate of Origin

E A Gorbunov et al.
Nutrition & diabetes, 5, e169-e169 (2015-07-07)
It has been previously shown that Subetta (a drug containing released-active forms of antibodies to the insulin receptor β-subunit and antibodies to endothelial nitric oxide synthase) stimulated insulin-induced adiponectin production by mature human adipocytes in the absence of insulin. Therefore...
Linda Jacobsen et al.
BioTechniques, 47(1), 617-624 (2009-07-15)
Transfection is used to introduce a gene of interest into a cell. To interpret the downstream results, understanding which effects are the true biological responses to the gene and which, if any, are off-target effects can be difficult. In order...
The insulin signaling system.
M F White et al.
The Journal of biological chemistry, 269(1), 1-4 (1994-01-07)
Renata M Martin et al.
Nature communications, 11(1), 2713-2713 (2020-06-03)
Despite their rapidly-expanding therapeutic potential, human pluripotent stem cell (hPSC)-derived cell therapies continue to have serious safety risks. Transplantation of hPSC-derived cell populations into preclinical models has generated teratomas (tumors arising from undifferentiated hPSCs), unwanted tissues, and other types of...
Marie S Isidor et al.
Adipocyte, 5(2), 175-185 (2016-07-08)
Brown adipose tissue is a promising therapeutic target for opposing obesity, glucose intolerance and insulin resistance. The ability to modulate gene expression in mature brown adipocytes is important to understand brown adipocyte function and delineate novel regulatory mechanisms of non-shivering...

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