MIF Recombinant Rabbit Monoclonal Antibody [PSH06-49]
cat.: HA722674
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IF-Cell, FC
Clonality: Monoclonal
Clone number: PSH06-49
Form: Liquid
Storage condition: Shipped at 4℃. Store at +4℃ short term (1-2 weeks). It is recommended to aliquot into single-use upon delivery. Store at -20℃ long term.
Storage buffer: PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 12 kDa
Isotype: IgG
Immunogen: Recombinant protein within human MIF aa 1-115.
Positive control: Jurkat cell lysate, THP-1 cell lysate, HL-60 cell lysate, U-87 MG cell lysate, HEK-293 cell lysate, A549 cell lysate, HepG2 cell lysate, HUVEC cell lysate, Neuro-2a cell lysate, NIH/3T3 cell lysate, C6 cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, Jurkat.
Subcellular location: Secreted, Cytoplasm.
Recommended Dilutions:
  WB
  IF-Cell
  FC
1:2,000-1:10,000
1:100
1:1,000
Uniprot #: SwissProt: P14174 Human | P34884 Mouse | P30904 Rat
Alternative names: GIF GLIF Glycosylation inhibiting factor Glycosylation-inhibiting factor L-dopachrome isomerase L-dopachrome tautomerase Macrophage migration inhibitory factor (glycosylation-inhibiting factor) Macrophage migration inhibitory factor MIF MIF protein MIF_HUMAN MMIF Phenylpyruvate tautomerase
Images
HA722674_1.jpg Fig1: Western blot analysis of MIF on different lysates with Rabbit anti-MIF antibody (HA722674) at 1/2,000 dilution.

Lane 1: Jurkat cell lysate (20 µg/Lane)
Lane 2: THP-1 cell lysate (20 µg/Lane)
Lane 3: HL-60 cell lysate (20 µg/Lane)
Lane 4: U-87 MG cell lysate (20 µg/Lane)
Lane 5: HEK-293 cell lysate (20 µg/Lane)
Lane 6: A549 cell lysate (20 µg/Lane)
Lane 7: HepG2 cell lysate (20 µg/Lane)
Lane 8: HUVEC cell lysate (20 µg/Lane)
Lane 9: Neuro-2a cell lysate (20 µg/Lane)
Lane 10: NIH/3T3 cell lysate (20 µg/Lane)
Lane 11: C6 cell lysate (20 µg/Lane)
Lane 12: Mouse brain tissue lysate (40 µg/Lane)
Lane 13: Rat brain tissue lysate (40 µg/Lane)

Predicted band size: 12 kDa
Observed band size: 12 kDa

Exposure time: 6 seconds; ECL: K1801;
4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722674) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA722674_2.jpg Fig2: Immunocytochemistry analysis of Jurkat cells labeling MIF with Rabbit anti-MIF antibody (HA722674) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-MIF antibody (HA722674) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. Nuclear DNA was labelled in blue with DAPI.

Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.
HA722674_3.jpg Fig3: Western blot analysis of MIF on different lysates with Rabbit anti-MIF antibody (HA722674) at 1/2,000 dilution.

Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-MIF KD cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 12 kDa
Observed band size: 12 kDa

Exposure time: 6 seconds; ECL: K1801;

4-20% SDS-PAGE gel.

Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA722674) at 1/2,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HA722674_4.jpg Fig4: Flow cytometric analysis of Jurkat cells labeling MIF.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA722674, 1/1,000) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.