HA722541

PHAPI2 / APRIL Recombinant Rabbit Monoclonal Antibody [JE35-49]

cat.: HA722541

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human
Applications:	WB, IF-Cell, IHC-P, FC
Clonality:	Monoclonal
Clone number:	JE35-49

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: 29 kDa
Isotype:	IgG
Immunogen:	Synthetic peptide.
Positive control:	293T cell lysate, Jurkat cell lysate, LNCaP cell lysate, Jurkat, human colon cancer tissue, human liver cancer tissue, human prostate tissue, human spleen tissue.
Subcellular location:	Nucleus, Cytoplasm.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:1,000 1:250 1:1,000 1:1,000
Uniprot #:	SwissProt: Q92688 Human
Alternative names:	Acidic (leucine rich) nuclear phosphoprotein 32 family member B Acidic leucine rich nuclear phosphoprotein 32 family member B Acidic leucine-rich nuclear phosphoprotein 32 family member B Acidic nuclear phosphoprotein 32 family member B Acidic protein rich in leucines AN32B_HUMAN ANP 32B ANP32B APRIL OTTHUMP0000006377 PAL 31 PAL31 PHAPI 2 PHAPI2 PHAPI2 protein PHAPI2a Proliferation related acidic leucine rich protein Putative HLA-DR-associated protein I-2 Silver stainable protein SSP 29 Silver stainable protein SSP29 Silver-stainable protein SSP29 Ssp 29 Ssp29

Images

Fig1: Western blot analysis of PHAPI2 / APRIL on different lysates with Rabbit anti-PHAPI2 / APRIL antibody (HA722541) at 1/1,000 dilution.

Lane 1: 293T cell lysate
Lane 2: Jurkat cell lysate
Lane 3: LNCaP cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 29 kDa
Observed band size: 29 kDa

Exposure time: 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 (HA722541) 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: Immunocytochemistry analysis of Jurkat cells labeling PHAPI2 / APRIL with Rabbit anti-PHAPI2 / APRIL antibody (HA722541) 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-PHAPI2 / APRIL antibody (HA722541) 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.

	Fig3: Immunohistochemical analysis of paraffin-embedded human colon cancer tissue with Rabbit anti-PHAPI2 / APRIL antibody (HA722541) 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 (HA722541) 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.
	Fig4: Immunohistochemical analysis of paraffin-embedded human liver cancer tissue with Rabbit anti-PHAPI2 / APRIL antibody (HA722541) 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 (HA722541) 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.
	Fig5: Immunohistochemical analysis of paraffin-embedded human prostate tissue with Rabbit anti-PHAPI2 / APRIL antibody (HA722541) 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 (HA722541) 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.

Fig6: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-PHAPI2 / APRIL antibody (HA722541) 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 (HA722541) 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: Flow cytometric analysis of Jurkat cells labeling PHAPI2 / APRIL.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA722541, 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”.