Recombinant Mouse EGF (R&D Cat# 2028-EG)
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INDEX CYTOKINE (Cat #) PAGE # HUMAN CYTOKINES 1 Endothelial cell growth supplement (ECGS) BD Biosciences Cat # 1 356006 Recombinant human /mouse/rat Activin A (R&D 338-AC) 1 Recombinant human Activin B (R&D Cat # 659-AB) 1 Recombinant human BMP4 (R&D 314BP) 2 Recombinant human BMPR-1A/ALK-3/Fc Chimera (R&D Cat # 2 315-BR) Recombinant human BMPR-1B/ALK-6/Fc Chimera (R&D Cat # 2 315-BR) Recombinant human Dkk1 (R&D Cat # 1096-DK) 3 Recombinant human EGF (R&D Cat# 236-EG) 3 Recombinant human EPO (Amgen–Sinai Pharmacy) 3 Recombinant human FGF2 (R&D 233-FB) 4 Recombinant human FGF4 (R&D Cat # 235-F4) 4 Recombinant human GM-CSF (R&D Cat # 215-GM) 5 Recombinant human HGF (R&D Cat # 294-HG) 5 Recombinant human IGF-1 (R&D 291-G1) 5 Recombinant human IGF-II (R&D Cat # 292-G2) 6 Recombinant human IL-3 (R&D Cat # 203-IL) 6 Recombinant human IL-6 (R&D 206-IL) 6 Recombinant human IL-11 (R&D Cat # 218-IL) 7 Recombinant human M-CSF (R&D Cat # 216-MC) 7 Recombinant human Netrin (R&D Cat # 1254-N4) 7 Recombinant human Osteopontin (R&D Cat # 1433-OP) 8 Recombinant human SCF (R&D 255-SC) 8 Recombinant human Sonic Hedgehog (R&D Cat # 1845-SH) 8 Recombinant human TPO (R&D Cat # 288-TP) 9 Recombinant human VEGF (R&D Cat # 293-VE) 9 MOUSE CYTOKINES Recombinant mouse DLL4 (R&D Cat # 1389-D4) 10 Recombinant mouse EGF (R&D Cat# 2028-EG) 10 Recombinant mouse GM-CSF (R&D Cat # 415-ML) 10 Recombinant mouse IGF-I (R&D Cat # 791-MG) 11 Recombinant mouse IL-11 (R&D Cat # 418-ML) 11 Recombinant mouse LIF (R&D Cat# 449-L) 11 Recombinant mouse M-CSF (R&D Cat # 416-ML) 12 Recombinant mouse Sonic Hedgehog (R&D Cat # 461-SH) 12 Recombinant mouse TPO (R&D Cat # 488-TO) 12 Recombinant mouse Wnt 3A (R&D Cat # 1324-WN/CF) 13 Recombinant mouse Wnt 5A (R&D Cat # 645-WN) 13 RECONSTITUTING CYTOKINES
1 Endothelial cell growth supplement (ECGS) BD Biosciences Cat # 356006 – 100mg/vial.
In a 14 ml tube, add 10 ml of IMDM reconstituted with glutamine and P/S.
Using 5 ml pipette, transfer ` 2-3 ml of this media into the vial containing lyophilized ECGS.
Let it dissolve at RT for 30 minutes. Mix gently.
Aliquot 2000uL / 5 ml tube (5 tubes). Label tubes with date and concentration of ECGS (10mg/ml ECGS).
Store aliquots at -20.
Recombinant human/mouse/rat Activin A (R&D Cat # 338-AC) – 250ug
In a 14 ml tube, add 5 ml of 0.1% BSA in 1X PBS previously filtered.
Using 10ml pipette, transfer some (`1ml) of this media into the vial containing lyophilized hActivin A
Let it dissolve at RT for 30 minutes. Transfer media back into 14ml tube containing the remaining 4 ml of media and mix gently with pipette.
Aliquot 500uL / eppendorf (10 eppendorfs). Label eppendorfs with date and concentration of hActA (50ug/ml hActA).
Store at –20.
Recombinant human Activin B (R&D Cat # 659-AB) – 250ug (NOT IN STOCK)
In a 14 ml tube, add 5 ml of 0.1% BSA in 1X PBS, previously filtered.
Using 10ml pipette, transfer some (`1ml) of this media into the vial containing lyophilized hActivinB
Let it dissolve at RT for 30 minutes. Transfer media back into 14ml tube containing the remaining 4 ml of media and mix gently with pipette.
Aliquot 500uL / eppendorf (10 eppendorfs). Label eppendorfs with date and concentration of hActB (50ug/ml hActB).
2 Store at –20.
Recombinant human BMP4 (R&D Cat# 314-BP) – 250ug Reagents: 10% BSA, 1N HCL and Sterile water
In a 50 ml tube, add 30ml of sterile water, 300uL of 10% BSA and 120uL of 1N HCL
Filter this mixture
Transfer 25ml of the above media into another 50ml tube.
Using 10ml pipette, transfer some of this media (2-5ml) into the vial containing lyophilized hBMP4
Let it dissolve at RT for 30 minutes. Transfer media back into 50 ml tube containing the remaining 20-23 ml of media and mix gently with pipette.
Aliquot 500uL / eppendorf (50 eppendorfs). Label eppendorfs with date and concentration of hBMP4 (10ug/ml hBMP4).
Store at –20.
Recombinant human BMPR-1A/ALK-3/Fc Chimera (R&D Cat # 315-BR) – 100ug
Using a 1 ml pipette, transfer 1ml of 0.1% BSA in 1X PBS, previously filtered into the vial containing lyophilized hBMPR-1A.
Let it dissolve at RT for 30 minutes. Mix gently.
Aliquot 100uL / eppendorf (10 eppendorfs). Label eppendorfs with date and concentration of hBMPR-1A (100ug / ml hBMPR-1A). Store at –20.
Recombinant human BMPR-1B/ALK-6/Fc Chimera (R&D Cat # 505-PR) – 100ug
Using a 1 ml pipette, transfer 1ml of 0.1% BSA in 1X PBS, previously filtered into the vial containing lyophilized hBMPR-1B.
Let it dissolve at RT for 30 minutes. Mix gently.
3 Aliquot 100uL / eppendorf (10 eppendorfs). Label eppendorfs with date and concentration of hBMPR-1B (100ug /ml BMPR-1B). Store at –20.
Recombinant human Dkk1 (R&D Cat # 1096-DK) – 250ug
In a 5 ml tube, add 2.5 ml of 0.1% BSA in 1X PBS, previously filtered.
Using 5 ml pipette, transfer media into the vial containing lyophilized hDKK1.
Let it dissolve at RT for 30 minutes. Mix gently.
Aliquot 200uL / eppendorf (13 eppendorfs). Label eppendorfs with date and concentration of hDKK1 (100ug /ml hDkk1). Store at –20.
Recombinant human EGF (R&D Cat# 236-EG) – 250ug Reagents: 1M acetic acid and 0.1% BSA in 1XPBS
In a 50 ml tube, add 30ml of 1X PBS containing 0.1% BSA.
To this media add 300ul (10uL/ml) of a 1M acetic acid solution.
Filter this mixture
Transfer 25ml of the above media into another 50ml tube.
Using 10ml pipette, transfer some of this media (2-5ml) into the vial containing lyophilized hEGF
Let it dissolve at RT for 30 minutes. Transfer media back into 50 ml tube containing the remaining 20-23 ml of media and mix gently with pipette.
Aliquot 1000uL / eppendorf (25 eppendorfs). Label eppendorfs with date and concentration of hEGF (10ug/ml hEGF).
Store at –20.
Recombinant human EPO (Amgen – Sinai Pharmacy) – 10,000U/ml
In a 14 ml tube, add 4 ml of 0.1% BSA in 1X PBS, previously filtered.
Remove metal cap. Insert sterile needle attached to a sterile 1ml syringe into the rubber cap of the vial.
4 Aspirate out the 1ml of hEPO and add it to the 4ml of 0.1% BSA in 1X PBS. Mix gently.
Aliquot 1000uL / eppendorf (5 eppendorfs). Label eppendorfs with date and concentration of hEPO (2000U/ml hEPO).
Store at 4 degrees. Do not freeze.
Recombinant human bFGF2 (R&D Cat # 233-FB) – 250ug
In a 50 ml tube, add 30ml of 0.1% BSA in 1X PBS
To the above media, add 300uL of 0.1M DTT
Filter this mixture
Transfer 25ml of the above media into another 50ml tube.
Using 10ml pipette, transfer some of this media (2-5ml) into the vial containing lyophilized human bFGF
Let it dissolve at RT for 30 minutes. Transfer media back into 50 ml tube containing the remaining 20-23 ml of media and mix gently with pipette.
Aliquot 1000uL / eppendorf (25 eppendorfs). Label eppendorfs with date and concentration of human bFGF (10ug/ml bFGF).
Store at –20.
Recombinant human FGF4 (R&D Cat # 235-F4) – 25ug
In a 5 ml tube, add 1 ml of 0.1% BSA in 1X PBS, previously filtered.
Transfer the 1 ml of the above media into another the lyophilized FGF4 vial.
Let it dissolve at RT for 30 minutes. Mix gently with pipette.
Aliquot 200uL / eppendorf (5 eppendorfs). Label eppendorfs with date and concentration of bFGF4 (10ug/ml FGF4).
Recombinant human GM-CSF (R&D Cat # 215-GM) – 10ug
5 In a 14 ml tube, add 10 ml of 0.1% BSA in 1X PBS
Using 10ml pipette, transfer some of this media (1-2 ml) into the vial containing lyophilized hGM-CSF
Let it dissolve at RT for 30 minutes. Transfer media back into 14 ml tube containing the remaining 9-8 ml of media and mix gently with pipette.
Aliquot 500uL / eppendorf (20 eppendorfs). Label eppendorfs with date and concentration of hGM-CSF (1 ug / ml bFGF).
Recombinant human HGF (R&D Cat # 294-HG) – 250ug
In a 5 ml tube, add 2.5 ml of 0.1% BSA in 1X PBS, previously filtered.
Using 5 ml pipette, transfer all this media into the vial containing lyophilized hHGF.
Let it dissolve at RT for 30 minutes. Mix gently.
Aliquot 200uL / eppendorf (13 eppendorfs). Label eppendorfs with date and concentration of hHGF (100ug / ml hHGF). Store at –20.
Recombinant human IGF-I (R&D Cat # 291-G1) – 250ug
In a 14 ml tube, add 10 ml of 0.1% BSA in 1X PBS, previously filtered.
Using 10ml pipette, transfer some (1-2ml) of this media into the vial containing lyophilized hIGF-I.
Let it dissolve at RT for 30 minutes. Transfer media back into 14 ml tube containing the remaining 8-10 ml of media and mix gently with pipette.
Aliquot 500uL / eppendorf (20 eppendorfs). Label eppendorfs with date and concentration of hIGF-I (25ug/ml hIGF-I).
Store at –20.
Recombinant human IGF-II (R&D Cat # 292-G2) – 25 ug
6 In a 5 ml tube, add 2.5 ml of 0.1% BSA in 1X PBS, previously filtered.
Using 1 ml pipette, transfer some (1-2ml) of this media into the vial containing lyophilized hIGF-II.
Let it dissolve at RT for 30 minutes. Transfer media back into 5 ml tube containing the remaining 0.5-1.5 ml of media and mix gently with pipette.
Aliquot 250uL / eppendorf (10 eppendorfs). Label eppendorfs with date and concentration of hIGF-II (10 ug/ml hIGF-II).
Store at –20.
Recombinant human IL-3 (R&D Cat # 203-IL) – 50 ug
In a 14 ml tube, add 5 ml of 0.1% BSA in 1X PBS, previously filtered.
Using 1 ml pipette, transfer some (1-2ml) of this media into the vial containing lyophilized hIL-3.
Let it dissolve at RT for 30 minutes. Transfer media back into 14 ml tube containing the remaining 3-4 ml of media and mix gently with pipette.
Aliquot 500 uL / eppendorf (10 eppendorfs). Label eppendorfs with date and concentration of hIL-3 (10 ug /ml, hIL-3).
Store at –20.
Recombinant human IL-6 (R&D Cat # 206-IL) – 50ug
In a 14 ml tube, add 5 ml of 0.1% BSA in 1X PBS, previously filtered.
Using 10ml pipette, transfer 1ml of this media into the vial containing lyophilized hIL-6
Let it dissolve at RT for 30 minutes. Transfer media back into 14ml tube containing the remaining 4 ml of media and mix gently with pipette.
Aliquot 500uL / eppendorf (10 eppendorfs). Label eppendorfs with date and concentration of hIL-6 (10ug/ml, hIL-6). Store at –20.
Recombinant human IL-11 (R&D Cat # 218-IL) – 250ug
7 In a 14 ml tube, add 5 ml of 0.1% BSA in 1X PBS, previously filtered.
Using 10ml pipette, transfer 1ml of this media into the vial containing lyophilized hIL-11.
Let it dissolve at RT for 30 minutes. Transfer media back into 14ml tube containing the remaining 4 ml of media and mix gently with pipette.
Aliquot 500uL / eppendorf (10 eppendorfs). Label eppendorfs with date and concentration of hIL-11 (50ug/ml, hIL-11). Store at –20.
Recombinant human M-CSF (R&D Cat # 216-MC) – 25ug
In a 5 ml tube, add 2.5 ml of 0.1% BSA in 1X PBS, previously filtered.
Using 1 ml pipette, transfer some of this media (1-2 ml) into the vial containing lyophilized hM-CSF.
Let it dissolve at RT for 30 minutes. Transfer media back into 5 ml tube containing the remaining 0.5-1.5 ml of media and mix gently with pipette.
Aliquot 500uL / eppendorf (5 eppendorfs). Label eppendorfs with date and concentration of hM-CSF (10 ug /ml hM-CSF).
Recombinant human Netrin (R&D Cat # 1254-N4) – 25 ug
In a 5 ml tube, add 2.5 ml of 0.1% BSA in 1X PBS, previously filtered.
Using 1 ml pipette, transfer 1 ml of this media into the vial containing lyophilized hNetrin.
Let it dissolve at RT for 30 minutes. Transfer media back into 5 ml tube containing the remaining 1.5 ml of media and mix gently with pipette.
Aliquot 250 uL / eppendorf (10 eppendorfs). Label eppendorfs with date and concentration of hNetrin (10 ug /ml hNetrin).
Store at –20.
Recombinant human Osteopontin (R&D Cat # 1433-OP) – 50 ug
8 In a 14 ml tube, add 5 ml of 0.1% BSA in 1X PBS, previously filtered.
Using 1 ml pipette, transfer 1 ml of this media into the vial containing lyophilized hOsteopontin.
Let it dissolve at RT for 30 minutes. Transfer media back into 14 ml tube containing the remaining 4 ml of media and mix gently with pipette.
Aliquot 500 uL / eppendorf (10 eppendorfs). Label eppendorfs with date and concentration of hOsteopontin (10 ug /ml hOsteopontin).
Store at –20.
Recombinant human SCF (R&D Cat # 255-SC) – 50ug
In a 5 ml tube, add 1 ml of 0.1% BSA in 1X PBS, previously filtered.
Using 1 ml p1000, transfer 1ml of this media into the vial containing lyophilized hSCF.
Let it dissolve at RT for 30 minutes. Mix gently.
Aliquot 500uL / eppendorf (2 eppendorfs). Label eppendorfs with date and concentration of hSCF (50ug/ml hSCF).
Store at –20.
Recombinant human Sonic Hedgehog (R&D Cat # 1845-SH) – 50 ug
Using p1000, transfer 250ul of 1X PBS containing 0.1% BSA into the vial containing lyophilized hShh.
Let it dissolve at RT for 30 minutes and mix gently with pipette.
Aliquot 50 uL / eppendorf (5 eppendorfs). Label eppendorfs with date and concentration of hShh (200 ug/ml hShh).
Store at –20.
Recombinant human TPO (R&D Cat # 288-TP) – 250 ug
9 In a 50 ml tube, add 25ml of 0.1% BSA in 1X PBS, previously filtered.
Using 10ml pipette, transfer some of this media (2-5ml) into the vial containing lyophilized TPO
Let it dissolve at RT for 30 minutes. Transfer media back into 50 ml tube containing the remaining 20-23 ml of media and mix gently with pipette.
Aliquot 1000uL / eppendorf (25 eppendorfs). Label eppendorfs with date and concentration of hTPO (10ug/ml hTPO).
Recombinant human VEGF 165 (R&D Cat # 293-VE) – 50 ug
In a 14 ml tube, add 10 ml of 0.1% BSA in 1X PBS, previously filtered.
Using 10ml pipette, transfer some (1-2ml) of this media into the vial containing lyophilized hVEGF.
Let it dissolve at RT for 30 minutes. Transfer media back into 14 ml tube containing the remaining 8-10 ml of media and mix gently with pipette.
Aliquot 500uL / eppendorf (20 eppendorfs). Label eppendorfs with date and concentration of hVEGF (5ug/ml hVEGF). Store at –20.
10 MOUSE CYTOKINES
Recombinant mouse DLL4 (R&D Cat # 1389-D4) – 50 ug
In a 5 ml tube, add 2.5 ml of 0.1% BSA in 1X PBS, previously filtered.
Using a p1000, transfer 1 ml of this media into the vial containing lyophilized mDLL4.
Let it dissolve at RT for 30 minutes. Transfer media back into 5 ml tube containing the remaining 1.5 ml of media and mix gently with pipette.
Aliquot 500uL / eppendorf (5 eppendorfs). Label eppendorfs with date and concentration of mDLL4 (20ug/ml mDLL4).
Recombinant mouse EGF (R&D Cat# 2028-EG) – 200ug
Add 2 ml of 1X PBS containing 0.1% BSA to the vial containing lyophilized mEGF.
Let it dissolve at RT for 30 minutes. Mix gently.
Aliquot 500 uL / eppendorf (4 eppendorfs). Label eppendorfs with date and concentration of mEGF (100ug/ml mEGF).
Store at –20.
Recombinant mouse GM-CSF (R&D Cat # 415-ML) – 25ug
In a 5 ml tube, add 2.5 ml of 0.1% BSA in 1X PBS, previously filtered.
Using a p1000, transfer 1 ml of this media into the vial containing lyophilized mGM-CSF.
Let it dissolve at RT for 30 minutes. Transfer media back into 5 ml tube containing the remaining 1.5 ml of media and mix gently with pipette.
Aliquot 500uL / eppendorf (5 eppendorfs). Label eppendorfs with date and concentration of mGM-CSF (10 ug/ml mGM-CSF).
11 Recombinant mouse IGF-I (R&D Cat # 791-MG) – 50 ug
In a 5 ml tube, add 2 ml of 0.1% BSA in 1X PBS, previously filtered.
Using a p1000 pipette, transfer 1ml of this media into the vial containing lyophilized mIGF-I.
Let it dissolve at RT for 30 minutes. Transfer media back into 5 ml tube containing the remaining 1 ml of media and mix gently with pipette.
Aliquot 250uL / eppendorf (8 eppendorfs). Label eppendorfs with date and concentration of mIGF-I (25ug/ml mIGF-I). Use at 1uL/ml.
Store at –20.
Recombinant mouse IL-11 (R&D Cat # 418-ML) – 25 ug
In a 14 ml tube, add 5 ml of 0.1% BSA in 1X PBS, previously filtered.
Using a p1000 pipette, transfer 1ml of this media into the vial containing lyophilized mIL-11.
Let it dissolve at RT for 30 minutes. Transfer media back into 14 ml tube containing the remaining 4 ml of media and mix gently with pipette.
Aliquot 500uL / eppendorf (10 eppendorfs). Label eppendorfs with date and concentration of mIL-11 (5 ug/ml mIL-11). Use at 1uL/ml.
Store at –20.
Recombinant mouse LIF (R&D Cat# 449-L) – 50ug
In a 14 ml tube, add 5 ml of 0.1% BSA in 1X PBS, previously filtered.
Using 1 ml pipette, transfer 1 ml into the vial containing lyophilized mLIF.
Let it dissolve at RT for 30 minutes. Transfer media back into 14 ml tube containing the remaining 4 ml of media and mix gently with pipette.
Aliquot 500uL / eppendorf (10 eppendorfs). Label eppendorfs with date and concentration of mLIF (10ug/ml mLIF). Use @ 1uL/ml for mESC maintenance in serum free media. Store at –20.
12 Recombinant mouse M-CSF (R&D Cat # 416-ML) – 50ug
In a 14 ml tube, add 5 ml of 0.1% BSA in 1X PBS, previously filtered.
Using 1 ml pipette, transfer 1 ml) into the vial containing lyophilized mM-CSF.
Let it dissolve at RT for 30 minutes. Transfer media back into 14 ml tube containing the remaining 4 ml of media and mix gently with pipette.
Aliquot 500uL /eppendorf (10 eppendorfs). Label eppendorfs with date and concentration of mM-CSF (10ug/ml mM-CSF).
Recombinant mouse Sonic Hedgehog (R&D Cat # 461-SH) – 25 ugs
Using p1000, transfer 500ul of 1X PBS containing 0.1% BSA into the vial containing lyophilized mShh.
Let it dissolve at RT for 30 minutes and mix gently with pipette.
Aliquot 100 uL /eppendorf (5 eppendorfs). Label eppendorfs with date and concentration of mShh (50 ug/ml mShh).
Store at –20.
Recombinant mouse TPO (R&D Cat # 488-TO) – 50 ug
In a 50 ml tube, add 5ml of 0.1% BSA in 1X PBS, previously filtered.
Using 1 ml pipette, transfer 1 ml into the vial containing lyophilized mTPO
Let it dissolve at RT for 30 minutes. Transfer media back into 5 ml tube containing the remaining 4 ml of media and mix gently with pipette.
Aliquot 500uL /eppendorf (10 eppendorfs). Label eppendorfs with date and concentration of mTPO (10 ug /ml mTPO).
13 Recombinant mouse Wnt 3A (R&D Cat # 1324-WN/CF)
Supplied as a 0.2uM filtered 5ml solution in 1X PBS containing 0.1mM EDTA, 0.5% CHAPS, pH 6.8 containing 200ug of Wnt3A @ a concentration of 40ng/ml.
Using p1000 pipette, aliquot 200uL / eppendorf (25 eppendorfs). Label eppendorfs with date and concentration of mWnt3A (40 ng/ml mWnt3A).
Recombinant mouse Wnt 5A (R&D Cat # 645-WN) – 10 ug
Using p1000 pipette, transfer 1ml of 1X PBS with 0.1% BSA, previously filtered into the lyophilized vial containing mWnt5A.
Let it dissolve at RT for 30 minutes.
Aliquot 100uL / eppendorf (10 eppendorfs). Label eppendorfs with date and concentration of mWnt5A (10 ug / ml mWnt5A).
14