HA500525

WTAP Rabbit Polyclonal Antibody

cat.: HA500525

Product Type:	Rabbit polyclonal IgG, primary antibodies
Species reactivity:	Human, Mouse, Rat
Applications:	WB, IHC-P, IF-Cell
Clonality:	Polyclonal

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	PBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2ug/ul
Purification:	Immunogen affinity purified.
Molecular weight:	Predicted band size: 44 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human WTAP aa 1-300 / 396.
Positive control:	K562 cell lysate, Jurkat cell lysate, Hela cell lysate, HepG2 cell lysate, 293T cell lysate, NIH/3T3 cell lysate, PC-12 cell lysate, human breast carcinoma tissue, human colon tissue, mouse small intestine tissue, rat small intestine tissue, NIH/3T3.
Subcellular location:	Nucleus speckle, Nucleus, nucleoplasm, Cytoplasm.
Recommended Dilutions: WB IHC-P IF-Cell	1:1,000 1:2,000 1:200
Uniprot #:	SwissProt: Q15007 Human \| Q9ER69 Mouse Entrez Gene: 499020 Rat
Alternative names:	DKFZp686F20131 Female-lethal(2)D homolog FL2D_HUMAN hFL(2)D KIAA0105 MGC3925 Mum2 OTTHUMP00000017522 OTTHUMP00000017523 PNAS 132 PNAS-132 PNAS132 Pre mRNA splicing regulator WTAP Pre-mRNA-splicing regulator WTAP Putative pre mRNA splicing regulator female lethal 2D homolog Putative pre mRNA splicing regulator female lethal(2D) Putative pre-mRNA splicing regulator female lethal(2D) homolog putative pre-mRNA splicing regulator female-lethal(2D) Wilms tumor 1 associated protein Wilms tumor 1 associating protein Wilms tumor 1-associating protein Wilms' tumor 1 associating protein Wilms' tumour 1-associating protein WT1 associated protein WT1-associated protein WT1-associating protein wtap

Images

Fig1: Western blot analysis of WTAP on different lysates with Rabbit anti-WTAP antibody (HA500525) at 1/1,000 dilution.

Lane 1: K562 cell lysate (12.5 µg/Lane)
Lane 2: Jurkat cell lysate (10 µg/Lane)
Lane 3: Hela cell lysate (10 µg/Lane)
Lane 4: HepG2 cell lysate (10 µg/Lane)
Lane 5: 293T cell lysate (10 µg/Lane)
Lane 6: NIH/3T3 cell lysate (10 µg/Lane)
Lane 7: PC-12 cell lysate (10 µg/Lane)

Predicted band size: 44 kDa
Observed band size: 55 kDa

Exposure time: 30 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 (HA500525) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:200,000 dilution was used for 1 hour at room temperature.

Fig2: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Rabbit anti-WTAP antibody (HA500525) at 1/2,000 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 (HA500525) at 1/2,000 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.

	Fig3: Immunohistochemical analysis of paraffin-embedded human colon tissue with Rabbit anti-WTAP antibody (HA500525) at 1/2,000 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 (HA500525) at 1/2,000 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded mouse small intestine tissue with Rabbit anti-WTAP antibody (HA500525) at 1/2,000 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 (HA500525) at 1/2,000 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded rat small intestine tissue with Rabbit anti-WTAP antibody (HA500525) at 1/2,000 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 (HA500525) at 1/2,000 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.

Fig6: Immunocytochemistry analysis of NIH/3T3 cells labeling WTAP with Rabbit anti-WTAP antibody (HA500525) at 10μg/mL dilution.

Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-WTAP antibody (HA500525) at 10μg/mL dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. 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.

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