HA722093

CD30 Recombinant Rabbit Monoclonal Antibody [PSH04-12]

cat.: HA722093

Product Type:	Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity:	Human
Applications:	WB, IF-Cell, IHC-P, IF-Tissue
Clonality:	Monoclonal
Clone number:	PSH04-12

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:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 64 kDa
Isotype:	IgG
Immunogen:	Recombinant protein within human CD30 aa 1-385 / 595.
Positive control:	HDLM-2 cell lysate, HUT 102 cell lysate, HDLM-2, human anaplastic large cell lymphoma tissue, human embryonal carcinoma tissue, human tonsil tissue.
Subcellular location:	Cell membrane; Cytoplasm.
Recommended Dilutions: WB IF-Cell IHC-P IF-Tissue	1:1,000 1:100 1:1,500 1:100
Uniprot #:	SwissProt: P28908 Human
Alternative names:	CD 30 CD30 CD30 antigen CD30L receptor Cytokine receptor CD30 D1S166E KI 1 KI 1 antigen Ki-1 antigen KI1 Lymphocyte activation antigen CD30 TNFRSF 8 Tnfrsf8 TNR8_HUMAN Tumor necrosis factor receptor superfamily member 8

Images

Fig1: Western blot analysis of CD30 on different lysates with Rabbit anti-CD30 antibody (HA722093) at 1/1,000 dilution.

Lane 1: HDLM-2 cell lysate
Lane 2: HUT 102 cell lysate
Lane 3: U-937 cell lysate (negative)
Lane 4: HL-60 cell lysate (negative)
Lane 5: Daudi cell lysate (negative)
Lane 6: HeLa cell lysate (negative)

Lysates/proteins at 20 µg/Lane.

Predicted band size: 64 kDa
Observed band size: 75-120 kDa

Exposure time: 1 minute 2 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 (HA722093) 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 HDLM-2 (positive) and HeLa (negative) labeling CD30 with Rabbit anti-CD30 antibody (HA722093) at 1/100 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-CD30 antibody (HA722093) at 1/100 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 anaplastic large cell lymphoma tissue with Rabbit anti-CD30 antibody (HA722093) at 1/1,500 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 (HA722093) at 1/1,500 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 embryonal carcinoma tissue with Rabbit anti-CD30 antibody (HA722093) at 1/1,500 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 (HA722093) at 1/1,500 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 tonsil tissue with Rabbit anti-CD30 antibody (HA722093) at 1/1,500 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 (HA722093) at 1/1,500 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: Immunofluorescence analysis of paraffin-embedded human embryonal carcinoma tissue labeling CD30 with Rabbit anti-CD30 antibody (HA722093) at 1/100 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 10% negative goat serum for 1 hour at room temperature, washed with PBS, and then probed with the primary antibody (HA722093, green) at 1/100 dilution overnight at 4 ℃, washed with PBS. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclei were counterstained with DAPI (blue).

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