Invitrogen

MIF Polyclonal Antibody

Advanced Verification

This Antibody was verified by Cell treatment to ensure that the antibody binds to the antigen stated.

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Questions & Answers

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MIF Antibody in Immunocytochemistry (ICC/IF) — Immunofluorescence analysis of Macrophage migration inhibitory factor was performed using 70% confluent log phase U-87 MG cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 15 minutes, and blocked with 2% BSA for 45 minutes at room temperature. The cells were labeled with MIF Polyclonal Antibody (Product # PA5-89525) at 1:100 dilution in 0.1% BSA, incubated at 4 degree celsius overnight and then labeled with Donkey anti-Rabbit IgG (H+L) Highly Cross-Adsorbed Secondary Antibody, Alexa Fluor Plus 488 (Product # A32790), (1:2000 dilution), for 45 minutes at room temperature (Panel a: Green). Nuclei (Panel b:Blue) were stained with ProLong™ Diamond Antifade Mountant with DAPI (Product # P36962). F-actin (Panel c: Red) was stained with Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic and secreted localization. Panel e represents control cells with no primary antibody to assess background. The images were captured at 60X magnification.

MIF Antibody in Immunocytochemistry (ICC/IF) — Immunocytochemistry-Immunofluorescence analysis of MIF was performed in A-549 cells using MIF Polyclonal Antibody (Product # PA5-89525).

MIF Antibody in Western Blot (WB) — Western Blot was performed using Anti-MIF Polyclonal Antibody (Product # PA5-89525) and a 15 kDa band corresponding to Macrophage migration inhibitory factor was observed across cell lines tested and was upregulated in U-87 MG upon VEGF treatment. Whole cell extracts (30 µg lysate) of THP-1 (Lane 1), THP-1 treated with PTI (1X, 4hrs) (Lane 2), U-937 (Lane 3), U-87 MG (Lane 4), U-87 MG treated with Rec. hVEGF (100 ng/mL, 48hrs) (Lane 5), Jurkat (Lane 6), HUVEC (Lane 7), RAW 264.7 (Lane 8), HEK-293 (Lane 9) were electrophoresed using NuPAGE™ 12% Bis-Tris Protein Gel (Product # NP0342BOX). Resolved proteins were then transferred onto a Nitrocellulose membrane (Product # IB23002) by iBlot® 2 Dry Blotting System (Product # IB21001). The Blot was probed with the primary antibody (1:1000 dilution) and detected by chemiluminescence with Goat anti-Rabbit IgG (H+L) Superclonal™ Recombinant Secondary Antibody, HRP (Product # A27036, 1:6000 dilution) using the iBright FL 1000 (Product # A32752). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).

MIF Antibody in Western Blot (WB) — Western Blot analysis of MIF in extracts of various cell lines using MIF Polyclonal Antibody (Product # PA5-89525) at a dilution of 1:1000. A HRP Goat Anti-Rabbit IgG (H+L) secondary antibody was used at a dilution of 1:10,000. Lysates/proteins: 25 µg per lane. Blocking buffer: 3% nonfat dry milk in TBST.

MIF Antibody — Altered expression of proteins upon cell treatment demonstrates antibody specificity. Western blot using MIF Polyclonal Antibody (Product # PA5-89525), shows increase in expression of MIF protein upon VEGF treatment in U-87 MG cell line. {TM}

Product Details

PA5-89525

Applications	Tested Dilution	Publications
Western Blot (WB)	1:500-1:2,000	-
Immunocytochemistry (ICC/IF)	1:10 - 1:100	-

Product Specifications
Species Reactivity	Human, Mouse, Rat
Host/Isotype	Rabbit / IgG
Class	Polyclonal
Type	Antibody
Immunogen	Recombinant fusion protein containing a sequence corresponding to amino acids 1-115 of human MIF View antigen
Conjugate	Unconjugated
Form	liquid
Concentration	0.87 mg/mL
Purification	Antigen affinity chromatography
Storage buffer	PBS, pH 7.3, with 50% glycerol
Contains	0.02% sodium azide
Storage conditions	-20° C, Avoid Freeze/Thaw Cycles
RRID	AB_2805629

Product Specific Information

Immunogen sequence: MPMFIVNTNV PRASVPDGFL SELTQQLAQA TGKPPQYIAV HVVPDQLMAF GGSSEPCALC SLHSIGKIGG AQNRSYSKLL CGLLAERLRI SPDRVYINYY DMNAANVGWN NSTFA; Positive Samples: HL-60, THP-1, SW620, Mouse kidney, Mouse lung, Mouse heart; Cellular Location: Cytoplasm

Target Information

MIF (macrophage migration inhibitory factor) was one of the first cytokine activities to be discovered and was initially described as a T cell-derived factor that inhibits the random migration of macrophages (Weiser 1989). Recently, MIF was rediscovered as a pituitary hormone that act as the counterregulatory hormone for glucocorticoid action within the immune system (Bernhagen 1993, Mitchell 1995). MIF was released from macrophages and T cells in response to physiological concentrations of glucocorticoids. The secreted MIF counter-regulates the immunosuppressive effects of steroids on immune cell activation and cytokine production (Bucala 1998). MIF plays a critical role in the host control of inflammation and immunity.

For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.

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Bioinformatics

Protein Aliases: Delayed early response protein 6; DER6; GIF; Glutathione-binding 13 kDa protein; Glycosylation-inhibiting factor; HGNC:7097; L-dopachrome isomerase; L-dopachrome tautomerase; Macrophage migration inhibitory factor; MGC107654; MIF; Phenylpyruvate tautomerase

Gene Aliases: GIF; GLIF; MIF; MMIF

UniProt ID: (Human) P14174, (Mouse) P34884, (Rat) P30904

Entrez Gene ID: (Human) 4282, (Mouse) 17319, (Rat) 81683

Molecular Function: decarboxylase

prostaglandin biosynthetic process negative regulation of mature B cell apoptotic process inflammatory response cell surface receptor signaling pathway cell aging cell proliferation negative regulation of gene expression positive regulation of protein kinase A signaling carboxylic acid metabolic process DNA damage response, signal transduction by p53 class mediator positive regulation of B cell proliferation positive regulation of lipopolysaccharide-mediated signaling pathway negative regulation of cellular protein metabolic process negative regulation of myeloid cell apoptotic process positive regulation of peptidyl-serine phosphorylation positive regulation of phosphorylation regulation of macrophage activation negative regulation of apoptotic process positive regulation of MAP kinase activity negative regulation of DNA damage response, signal transduction by p53 class mediator innate immune response positive regulation of fibroblast proliferation positive regulation of cytokine secretion positive regulation of peptidyl-tyrosine phosphorylation leukocyte migration positive chemotaxis positive regulation of prostaglandin secretion involved in immune response positive regulation of myeloid leukocyte cytokine production involved in immune response protein homotrimerization positive regulation of ERK1 and ERK2 cascade negative regulation of cell cycle arrest positive regulation of arachidonic acid secretion negative regulation of cell aging negative regulation of intrinsic apoptotic signaling pathway in response to DNA damage by p53 class mediator positive regulation of chemokine (C-X-C motif) ligand 2 production positive regulation of protein phosphorylation brain renin-angiotensin system immune system process positive regulation of acute inflammatory response positive regulation of adaptive immune response spermatogenesis positive regulation of smooth muscle cell migration regulation of cell proliferation drinking behavior positive regulation of potassium ion transport positive regulation of glycolytic process positive regulation of glucose import positive regulation of axon regeneration positive regulation of immune response hair follicle development brain development aging metabolic process response to heat response to mechanical stimulus response to hormone response to inorganic substance response to estradiol response to lipopolysaccharide response to progesterone response to menaquinone response to insulin response to vitamin E wound healing response to drug skin development response to glucocorticoid cellular response to hydrogen peroxide cellular response to hypoxia

Process(es)

dopachrome isomerase activity receptor binding cytokine activity cytokine receptor binding protein binding chemoattractant activity phenylpyruvate tautomerase activity isomerase activity protein complex binding