M1501-8

JAK2 Mouse Monoclonal Antibody [6-D3]

cat.: M1501-8

Product Type:	Mouse monoclonal IgG2b, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P
Clonality:	Monoclonal
Clone number:	6-D3

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: 130 kDa
Isotype:	IgG2b
Immunogen:	Recombinant protein within Human JAK2 aa 883-1,132 / 1,132.
Positive control:	A549 cell lysate, K-562 cell lysate, TF-1 cell lysate, Jurkat cell lysate, RAW264.7 cell lysate, C6 cell lysate, K-562, RAW264.7, human kidney tissue, human lung carcinoma tissue, mouse kidney tissue, rat kidney tissue.
Subcellular location:	Cytopasm, nucleus
Recommended Dilutions: WB IF-Cell IHC-P	1:2,000 1:50-1:100 1:200
Uniprot #:	SwissProt: O60674 Human
Alternative names:	JAK 2 JAK-2 JAK2 JAK2_HUMAN Janus Activating Kinase 2 Janus kinase 2 (a protein tyrosine kinase) Janus kinase 2 JTK 10 JTK10 kinase Jak2 OTTHUMP00000043260 THCYT3 Tyrosine protein kinase JAK2 Tyrosine-protein kinase JAK2

Images

Fig1: Western blot analysis of JAK2 on different lysates with Mouse anti-JAK2 antibody (M1501-8) at 1/2,000 dilution.

Lane 1: A549-si NT cell lysate
Lane 2: A549-si JAK2 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 130 kDa
Observed band size: 130 kDa

Exposure time: 45 seconds;

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 (M1501-8) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/100,000 dilution was used for 1 hour at room temperature.

Fig2: Western blot analysis of JAK2 on different lysates with Mouse anti-JAK2 antibody (M1501-8) at 1/1,000 dilution.

Lane 1: K-562 cell lysate
Lane 2: TF-1 cell lysate
Lane 3: Jurkat cell lysate
Lane 4: RAW264.7 cell lysate
Lane 5: C6 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 130 kDa
Observed band size: 130 kDa

Exposure time: 40 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 (M1501-8) 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.

Fig3: Immunocytochemistry analysis of K-562 cells labeling JAK2 with Mouse anti-JAK2 antibody (M1501-8) 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-JAK2 antibody (M1501-8) 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.

Fig4: Immunocytochemistry analysis of RAW264.7 cells labeling JAK2 with Mouse anti-JAK2 antibody (M1501-8) 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-JAK2 antibody (M1501-8) 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: Immunohistochemical analysis of paraffin-embedded human kidney tissue with Mouse anti-JAK2 antibody (M1501-8) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (M1501-8) 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.
	Fig6: Immunohistochemical analysis of paraffin-embedded human lung carcinoma tissue with Mouse anti-JAK2 antibody (M1501-8) at 1/1,000 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (M1501-8) at 1/1,000 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.
	Fig7: Immunohistochemical analysis of paraffin-embedded mouse kidney tissue with Mouse anti-JAK2 antibody (M1501-8) at 1/200 dilution. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (M1501-8) 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.

Fig8: Immunohistochemical analysis of paraffin-embedded rat kidney tissue with Mouse anti-JAK2 antibody (M1501-8) at 1/200 dilution.

The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 2 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 (M1501-8) 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”.