HA722137

p57 Kip2 Recombinant Rabbit Monoclonal Antibody [JE02-08]

cat.: HA722137

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IF-Cell, IHC-P
Clonality:	Monoclonal
Clone number:	JE02-08

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 32 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide within Human p57 Kip2 aa 31-80 / 316.
Positive control:	HeLa cell lysate, HeLa treated with 50nM dexamethasone for 16 hours cell lysate, SH-SY5Y cell lysate, K-562 cell lysate, 293T cell lysate, Mouse brain tissue lysate, Rat brain tissue lysate, HeLa cells treated with 50nM dexamethasone for 16 hours, human placenta tissue, human brain tissue, mouse brain tissue, rat brain tissue.
Subcellular location:	Nucleus.
Recommended Dilutions: WB IF-Cell IHC-P	1:1,000-1:5,000 1:250 1:200
Uniprot #:	SwissProt: P49918 Human \| P49919 Mouse Entrez Gene: 246060 Rat
Alternative names:	Beckwith Wiedemann syndrome BWCR BWS CDKI CDKN 1C CDKN1C CDN1C_HUMAN Cyclin dependent kinase inhibitor 1C Cyclin dependent kinase inhibitor p57 Cyclin-dependent kinase inhibitor 1C Cyclin-dependent kinase inhibitor p57 KIP 2 KIP2 p57 p57 Kip 2 p57KIP2 WBS

Images

Fig1: Western blot analysis of p57 Kip2 on different lysates with Rabbit anti-p57 Kip2 antibody (HA722137) at 1/1,000 dilution.

Lane 1: HeLa cell lysate (20 µg/Lane)
Lane 2: HeLa treated with 50nM dexamethasone for 16 hours cell lysate (20 µg/Lane)
Lane 3: SH-SY5Y cell lysate (20 µg/Lane)
Lane 4: K-562 cell lysate (20 µg/Lane)
Lane 5: 293T cell lysate (20 µg/Lane)
Lane 6: Mouse brain tissue lysate (40 µg/Lane)
Lane 7: Rat brain tissue lysate (40 µg/Lane)

Predicted band size: 32 kDa
Observed band size: 50 kDa

Exposure time: Lane 1-3: 59 seconds; Lane 4-7: 3 minutes; 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 (HA722137) at 1/1,000 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.

Fig2: Western blot analysis of p57 Kip2 on different lysates with Rabbit anti-p57 Kip2 antibody (HA722137) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HeLa treated with 50nM dexamethasone for 16 hours cell lysate
Lane 3: HeLa treated with 50nM dexamethasone for 16 hours cell lysate, then the membrane treated with λpp for 1 hour

Lysates/proteins at 15 µg/Lane.

Predicted band size: 32 kDa
Observed band size: 50 kDa

Exposure time: 59 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 (HA722137) at 1/5,000 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.

	Fig3: Immunocytochemistry analysis of HeLa cells treated with or without 50nM dexamethasone for 16 hours labeling p57 Kip2 with Rabbit anti-p57 Kip2 antibody (HA722137) at 1/250 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-p57 Kip2 antibody (HA722137) at 1/250 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human placenta tissue with Rabbit anti-p57 Kip2 antibody (HA722137) 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 (HA722137) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human brain tissue with Rabbit anti-p57 Kip2 antibody (HA722137) 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 (HA722137) 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 mouse brain tissue with Rabbit anti-p57 Kip2 antibody (HA722137) 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 (HA722137) 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.

Fig7: Immunohistochemical analysis of paraffin-embedded rat brain tissue with Rabbit anti-p57 Kip2 antibody (HA722137) 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 (HA722137) 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”.