M1301-1

Lin28A Mouse Monoclonal Antibody [A4-A6]

cat.: M1301-1

Product Type:	Mouse monoclonal IgG2b, primary antibodies
Species reactivity:	Human, Mouse
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Monoclonal
Clone number:	A4-A6

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4), 0.2% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 23 kDa
Isotype:	IgG2b
Immunogen:	Recombinant protein within human Lin28A aa 1-209/209.
Positive control:	NCCIT cell lysate, JAR cell lysate, F9 cell lysates, NCCIT, F9, mouse heart tissue.
Subcellular location:	Cytoplasm, Nucleus.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:1,000-1:2,000 1:100-1:250 1:200 1:1,000
Uniprot #:	SwissProt: Q9H9Z2 Human \| Q8K3Y3 Mouse
Alternative names:	CSDD1 FLJ12457 LIN 28 Lin 28 homolog A (C. elegans) Lin-28A LIN28 LIN28A LN28A_HUMAN Protein lin-28 homolog A ZCCHC1 zinc finger CCHC domain containing 1 Zinc finger CCHC domain-containing protein 1

Images

Fig1: Western blot analysis of Lin28A on different lysates with Mouse anti-Lin28A antibody (M1301-1) at 1/1,000 dilution.

Lane 1: NCCIT cell lysate
Lane 2: HeLa cell lysate (negative)
Lane 3: JAR cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 23 kDa
Observed band size: 28 kDa

Exposure time: Lane 1-2: 9 seconds; Lane 3: 3 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 (M1301-1) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of Lin28A on F9 cell lysates with Mouse anti-Lin28A antibody (M1301-1) at 1/2,000 dilution.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 23 kDa
Observed band size: 28 kDa

Exposure time: 10 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 (M1301-1) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

	Fig3: Immunocytochemistry analysis of NCCIT cells labeling Lin28A with Mouse anti-Lin28A antibody (M1301-1) at 1/250 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 Mouse anti-Lin28A antibody (M1301-1) at 1/250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.
	Fig4: Immunocytochemistry analysis of F9 cells labeling Lin28A with Mouse anti-Lin28A antibody (M1301-1) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 Mouse anti-Lin28A antibody (M1301-1) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.
	Fig5: Flow cytometric analysis of NCCIT cells labeling Lin28A. Cells were fixed and permeabilized. Then stained with the primary antibody (M1301-1, 1/1,000) (red) compared with Mouse 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-Mouse IgG Secondary antibody (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

Fig6: Immunohistochemical analysis of paraffin-embedded mouse heart tissue with Mouse anti-Lin28A antibody (M1301-1) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH₂O and PBS, and then probed with the primary antibody (M1301-1) at 1/200 dilution for 1 hour at room temperature. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.