HA601025

CD133 Mouse Monoclonal Antibody [A8C2]

cat.: HA601025

Product Type:	Mouse monoclonal IgG1, primary antibodies
Species reactivity:	Human
Applications:	WB, IHC-P, FC
Clonality:	Monoclonal
Clone number:	A8C2

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:	Protein G affinity purified.
Molecular weight:	Predicted band size: 97 kDa
Isotype:	IgG1
Immunogen:	Recombinant protein within human CD133 aa 151-400/865.
Positive control:	NCCIT cell lysates, human kidney tissue lysates, HT-29 cell lysates, HepG2 cell lysates, human colon carcinoma tissue, human breast tissue, human kidney tissue, human pancreas tissue, NCCIT.
Subcellular location:	Endoplasmic reticulum. Plasma membrane. Cell projection.
Recommended Dilutions: WB IHC-P FC	1:500 1:600 1:500-1:1,000
Uniprot #:	SwissProt: O43490 Human
Alternative names:	AC133 Antigen AC133 CD133 CORD12 Hematopoietic stem cell antigen hProminin MCDR2 MSTP061 OTTHUMP00000217744 OTTHUMP00000217745 OTTHUMP00000217746 PROM1 PROM1_HUMAN Prominin I Prominin like 1 Prominin like protein 1 precursor Prominin mouse like 1 Prominin-1 Prominin-like protein 1 Prominin1 PROML1 RP41 STGD4

Images

Fig1: Western blot analysis of CD133 on NCCIT cell lysates with Mouse anti-CD133 antibody (HA601025) at 1/500 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 97 kDa
Observed band size: 130 kDa

Exposure time: 2 minutes;

8% 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 (HA601025) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. 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 CD133 on human kidney tissue lysates with Mouse anti-CD133 antibody (HA601025) at 1/500 dilution.

Lysates/proteins at 20 µg/Lane.

Predicted band size: 97 kDa
Observed band size: 110 kDa

Exposure time: 30 seconds;

6% 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 (HA601025) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.

Fig3: Western blot analysis of CD133 on HT-29 cell lysates with Mouse anti-CD133 antibody (HA601025) at 1/500 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 97 kDa
Observed band size: 110 kDa

Exposure time: 1 minute;

6% 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 (HA601025) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.

Fig4: Western blot analysis of CD133 on HepG2 cell lysates with Mouse anti-CD133 antibody (HA601025) at 1/500 dilution.

Lysates/proteins at 10 µg/Lane.

Predicted band size: 97 kDa
Observed band size: 110 kDa

Exposure time: 2 minutes;

8% 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 (HA601025) at 1/500 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1:100,000 dilution was used for 1 hour at room temperature.

	Fig5: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue with Mouse anti-CD133 antibody (HA601025) at 1/600 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 (HA601025) at 1/600 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 breast tissue with Mouse anti-CD133 antibody (HA601025) at 1/600 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 (HA601025) at 1/600 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 human kidney tissue with Mouse anti-CD133 antibody (HA601025) at 1/600 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 (HA601025) at 1/600 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 human pancreas tissue with Mouse anti-CD133 antibody (HA601025) at 1/600 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 (HA601025) at 1/600 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.

Fig9: Flow cytometric analysis of NCCIT cells labeling CD133.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA601025, 1ug/ml) (red) compared with Mouse 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-Mouse IgG Secondary antibody 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”.