HA601583

Cytokeratin 7 Recombinant Antibody [ST50-05] - Mouse IgG1 (Chimeric)

cat.: HA601583

Product Type:	Recombinant Chimeric Antibody, primary antibodies
Species reactivity:	Human
Applications:	WB, IF-Cell, IHC-P, FC
Clone number:	ST50-05

Form:	Liquid
Storage condition:	Shipped at 4℃. Store at +4℃ short term (1-2 weeks). Store at -20℃ long term.
Storage buffer:	1*PBS (pH7.4), 0.1% BSA, 40% Glycerol, 0.2% Proclean 950.
Concentration:	1ug/ul
Purification:	Protein A affinity purified.
Molecular weight:	Predicted band size: 51 kDa
Immunogen:	Synthetic peptide within Human Cytokeratin 7 aa 18-67.
Positive control:	HeLa cell lysate, A549 cell lysate, SK-OV-3 cell lysate, human breast carcinoma tissue, human lung adenocarcinoma tissue, human liver tissue, HeLa.
Subcellular location:	Cytoplasm.
Recommended Dilutions: WB IF-Cell IHC-P FC	1:5,000 1:100 1:500-1:2,000 1:1,000
Uniprot #:	SwissProt: P08729 Human
Alternative names:	CK 7 CK-7 CK7 Cytokeratin 7 Cytokeratin-7 D15Wsu77e K2C7 K2C7_HUMAN K7 Keratin 7 Keratin 7, type II Keratin type II cytoskeletal 7 Keratin, 55K type II cytoskeletal Keratin, simple epithelial Keratin, simple epithelial type I, K7 Keratin, type II cytoskeletal 7 Keratin-7 Krt2-7 KRT7 MGC11625 MGC129731 MGC3625 Sarcolectin SCL Type II mesothelial keratin K7 Type-II keratin Kb7 Cytokeratin7

Images

Fig1: Western blot analysis of Cytokeratin 7 on different lysates with Mouse anti-Cytokeratin 7 antibody (HA601583) at 1/5,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: MCF7 cell lysate (negative)
Lane 3: A549 cell lysate
Lane 4: SK-OV-3 cell lysate

Lysates/proteins at 20 µg/Lane.

Predicted band size: 51 kDa
Observed band size: 51 kDa

Exposure time: 20 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 (HA601583) at 1/5,000 dilution was used in primary antibody dilution (K1803) at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.

Fig2: Immunocytochemistry analysis of HeLa (positive) and MCF7 (negative) labeling Cytokeratin 7 with Mouse anti-Cytokeratin 7 antibody (HA601583) at 1/100 dilution.

Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 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 Mouse anti-Cytokeratin 7 antibody (HA601583) at 1/100 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) 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 (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.

	Fig3: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue with Mouse anti-Cytokeratin 7 antibody (HA601583) at 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 (HA601583) at 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 lung adenocarcinoma tissue with Mouse anti-Cytokeratin 7 antibody (HA601583) at 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 (HA601583) at 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 liver tissue with Mouse anti-Cytokeratin 7 antibody (HA601583) at 1/2,000 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 (HA601583) 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: Immunohistochemical analysis of paraffin-embedded human colon carcinoma tissue (negative) with Mouse anti-Cytokeratin 7 antibody (HA601583) at 1/2,000 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 (HA601583) 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.

Fig7: Flow cytometric analysis of MCF7 (left, negative) and HeLa (right, positive) cells labeling Cytokeratin 7.

Cells were fixed and permeabilized. Then stained with the primary antibody (HA601583, 1/1,000) (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 (HA1125) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).

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