Molecular Characterization

Molecular Characterization: Recombinant hMCP-1 has a calculated MW of 8,685. DTT-reduced protein migrates as a 10 kDa polypeptide. The nonreduced protein migrates at 12 kDa. The expected amino-terminal QPDAI of recombinant hMCP-1 was verified by amino acid sequencing.

Purity

>95% as determined by SDS-PAGE of 6 μg reduced (+) and nonreduced (-) recombinant hMCP-1. All lots are greater than 95% pure.

Bioactivity

The activity of hMCP-1 was determined using a THP-1 cell migration assay. The ED₅₀ of each lot is between 1-11 ng/ml.

Bioactivity

hMCP-1-induced migration of THP-1 cells was assessed. THP-1 cells were incubated in a 96-well transwell plate with increasing concentrations of hMCP-1 in the bottom chamber. After 2 hr, the number of THP-1 cells that migrated to the bottom chamber of the transwell was quantified by measuring DNA content using a fluorescent dye.

Coomassie Gel

The purity of recombinant hMCP-1 was determined by SDS-PAGE of 6 µg reduced (+) and nonreduced (-) recombinant hMCP-1 and staining overnight with Coomassie Blue.

Applications

Optimal concentration for the desired application should be determined by the user.

Endotoxin

Less than 0.01 ng endotoxin/1 μg hMCP-1.

Formulation

With carrier: Lyophilized from a 0.22 μm filtered solution of hMCP-1 in 20 mM Tris, pH 7.2 containing 20 μg BSA per 1 μg hMCP-1. Carrier free: Lyophilized from a 0.22 μm filtered solution of hMCP-1 in 20 mM Tris, pH 7.2.

Background

MCP-1 (CCL2) is the first member of the C-C family of chemokines to be identified (1). C-C chemokines are characterized by two adjacent cysteine residues within the polypeptide, which form an intra-molecular disulfide bond. MCP-1 is a potent chemotactic factor for monocytes/macrophages, T cells and a subset of NK cells (1-4). The MCP-1 receptor, CCR2, is expressed as two splice isoforms, CCR2A and CCR2B, of which CCR2B is the predominant form (1). MCP-1 is secreted by adipocytes and appears to be one of many links between obesity, inflammation, and diabetes (1). MCP-1/CCR2 signaling appears to play a key role in γδ effector T cells recruitment and anti-tumor responses in a murine B16 melanoma model (2). Conversely, CCL2 expression is upregulated in many types of cancer and has been implicated in promoting tumor cell survival, proliferation, and tumor associated inflammation (4).

11946      Human TL1A/TNFSF15 (hTL1A)

11996      Mouse His6Interleukin-25/IL-17E (mHis6IL-25)

12296     Human Fibroblast Growth Factor 9 (hFGF9)

12337     Human His6Interleukin-25/IL-17E (hHis6IL-25)

12568     Human His6IGFBP2

12579     Human His6IGFBP3 (hHis6IGFBP3)

12670     MouseHis6Interleukin-21 (mHis6IL-21)

12786     Human Interleukin-6 Receptor α (hIL-6Rα)

12838     Mouse MCP-1

12900     Human MCP-1

3583       Human CD40 Ligand (hCD40L)

3897       Human Brain-Derived Neurotrophic Factor (BDNF)

4697       Human BMP2

4698       Mouse His6 Tumor Necrosis Factor-α (mHis6TNF-α)

5154       Human Latent Transforming Growth Factor β1 (hLatent TGF-β1)

5164       Human Interleukin-28A (hIL-28A/IFN-λ2)

5178       Mouse Tumor Necrosis Factor-α (mTNF-α)

5183       Human Interleukin-29 (hIL-29)

5191       Mouse Granulocyte Macrophage Colony Stimulating Factor (mGM-CSF)

5201       Mouse Interleukin-2 (mIL-2)

5204       Mouse Interleukin-1β (mIL-1β)

5208       Mouse Interleukin-4 (mIL-4)

5211       Mouse Vascular Endothelial Growth Factor-164 (mVEGF164 )

5216       Mouse Interleukin-6 (mIL-6)

5217       Mouse Interleukin-7 (mIL-7)

5218       Human Neuregulin-1 (hNRG-1)

5221       Human β-Nerve Growth Factor (hβ-NGF)

5222       Mouse Interferon-γ (mIFN-γ)

5223       Mouse Stem Cell Factor (mSCF)

5224       Mouse Interleukin-22 (mIL-22)

5227       Mouse Interleukin-17A (mIL-17A)

5228       Mouse Macrophage Colony Stimulating Factor (mM-CSF)

5231       Mouse Transforming Growth Factor β1 (mTGF-β1)

5233       Human BAFF/TNFSF13B (hBAFF)

5234       Human FGF acidic (hFGF acidic)

5235       Human Betacellulin (hBTC)

5236       Human Interleukin-1α (hIL-1α)

5237       Human Neurotrophin-3 (hNT-3)

5238       Human Insulin-like Growth Factor II (hIGF-II)

5242       Mouse Interleukin-13 (mIL-13)

5261       Mouse Interleukin-10 (mIL-10)

5273       Mouse Interleukin-1α (mIL-1α)

5312       Human RANKL/TRANCE/TNFSF11 (hRANKL)

5314       Mouse Vascular Endothelial Growth Factor-120 (mVEGF120 )

5331       Mouse Epidermal Growth Factor (mEGF)

5358       Human Interleukin-10 (hIL-10) (mammalian derived)

5367       Human Oncostatin M (hOSM)

5371       Mouse Oncostatin M (mOSM)

5413       Human His6BAFF/TNFSF13B (hHis6 BAFF)

5414       Mouse Basic Fibroblast Growth Factor (mFGF basic/FGF2)

5452       Human His6Fas Ligand/TNFSF6 (hHis6FasL)

5454       Human Interleukin-2 (hIL-2) (mammalian derived)

5493       Human Epigen

5494       Human Epiregulin

5495       Human Transforming Growth Factor α (hTGF-α)

5592       Human Neurotrophin-4 (hNT-4)

5712       Human Stromal Cell-derived Factor 1β/CXCL12 (hSDF1β)

5717       Human Cystatin C (hCystatin C)

5719       Human Leptin/OB (hLeptin)

5725       Mouse His6Interleukin-27 (mHis6IL-27)

5874       Rat Vascular Endothelial Growth Factor-164 (rVEGF164 )

5924       Human Interleukin-5 (hIL-5)

6980       Human Erythropoietin (hEPO)

6983       Mouse His6Interleukin-33 (mHis6IL-33)

8012       Human His6Thymic Stromal Lymphopoietin (hHis6TSLP)

8065       Human Vascular Endothelial Growth Factor-165 (hVEGF165 )

8170       Human Interleukin-7 (hIL-7)

8199       Mouse Lipocalin-2 (mLCN2)

8230       Human Lymphotoxin-α/TNF-β/TNFSF1 (hLT-α)

8406       Human Transforming Growth Factor β2 (hTGF-β2)

8425       Human Transforming Growth Factor β3 (hTGF-β3)

8445       Human C-C Motif Chemokine 3 (hCCL3/MIP-1-α)

8460       Human His64­1BB Ligand/TNFSF9 (hHis64-1BBL)

8796       Human Interleukin-28B (hIL-28B/IFN-λ3)

8846       Human Interleukin-17A/F Heterodimer (hIL-17A/F)

8857       Mouse His6Interleukin-6 (mHis6IL-6)

8900       Human Interleukin-1β (hIL-1β)

8901       Human Interferon-γ (hIFN-γ)

8902       Human Tumor Necrosis Factor-α (hTNF-α)

8903       Human Interleukin-10 (hIL-10)

8904       Human Interleukin-6 (hIL-6)

8905       Human Interleukin-13 (hIL-13)

8906       Human Interleukin-17F (hIL-17F)

8907       Human Interleukin-2 (hIL-2)

8908       Human Vascular Endothelial Growth Factor-121 (hVEGF121 )

8910       Human Basic Fibroblast Growth Factor (hFGF basic/FGF2)

8911       Human Leukemia Inhibitory Factor (hLIF)

8912       Human Platelet-Derived Growth Factor BB (hPDGF-BB)

8913       Human Platelet-Derived Growth Factor AA (hPDGF-AA)

8915       Human Transforming Growth Factor β1 (hTGF-β1)

8916       Human Epidermal Growth Factor (hEGF)

8917       Human Insulin-like Growth Factor I (hIGF-I)

8918       Human Interleukin-3 (hIL-3)

8919       Human Interleukin-4 (hIL-4)

8920       Human Interleukin-21 (hIL-21)

8921       Human Interleukin-8 (hIL-8)

8922       Human Granulocyte Macrophage Colony Stimulating Factor (hGM-CSF)

8923       Mouse Interleukin-3 (mIL-3)

8924       Human Fms-related Tyrosine Kinase 3 Ligand (hFLT3L)

8925       Human Stem Cell Factor (hSCF)

8927       Human Interferon-α1 (hIFN-α1)

8928       Human Interleukin-17A (hIL-17A)

8929       Human Macrophage Colony Stimulating Factor (hM-CSF)

8930       Human Granulocyte Colony Stimulating Factor (hG-CSF)

8931       Human Interleukin-22 (hIL-22)

9584       Mouse Interleukin-17B (mIL-17B)

9897       Mouse Insulin-like Growth Factor I (mIGF-I)