Cytokeratin 18 Mouse Monoclonal Antibody [A4B3]
cat.: HA600021
Product Type: Mouse monoclonal IgG2a, primary antibodies
Species reactivity: Human, Mouse, Rat
Applications: WB, IHC-P, FC, IF-Cell
Clonality: Monoclonal
Clone number: A4B3
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 1%BSA, 50%Glycerol. Preservative: 0.05% Sodium Azide.
Concentration: 2ug/ul
Purification: Protein G affinity purified.
Molecular weight: Predicted band size: 48 kDa
Isotype: IgG2a
Immunogen: Recombinant protein within human Cytokeratin 18 aa 1-150.
Positive control: SK-Br-3 cell lysate, A549 cell lysate, A431 cell lysate, MCF7 cell lysate, mouse liver tissue lysate, rat liver tissue lysate, HeLa, human liver tissue, human skin tissue, human prostate carcinoma tissue, human breast carcinoma tissue, MCF-7.
Subcellular location: Nucleus matrix, Cytoplasm, perinuclear region, Nucleus, nucleolus.
Recommended Dilutions:
  WB
  IHC-P
  FC
  IF-Cell
1:500-1:2,000
1:100-1:500
1:50-1:100
1:100
Uniprot #: SwissProt: P05783 Human | P05784 Mouse | Q5BJY9 Rat
Alternative names: Cell proliferation inducing gene 46 protein Cell proliferation inducing protein 46 Cell proliferation-inducing gene 46 protein CK 18 CK-18 CK18 CYK 18 CYK18 Cytokeratin 18 Cytokeratin endo B Cytokeratin-18 K 18 K18 K1C18_HUMAN KA18 Keratin 18 Keratin 18, type I Keratin D keratin, type I cytoskeletal 18 Keratin-18 Krt18
Images
HA600021_1.jpg Fig1: Western blot analysis of Cytokeratin 18 on different lysates with Mouse anti-Cytokeratin 18 antibody (HA600021) at 1/1,000 dilution.

Lane 1: SK-Br-3 cell lysate (20 µg/Lane)
Lane 2: A549 cell lysate (20 µg/Lane)
Lane 3: A431 cell lysate (20 µg/Lane)
Lane 4: MCF7 cell lysate (20 µg/Lane)
Lane 5: Mouse liver tissue lysate (40 µg/Lane)
Lane 6: Rat liver tissue lysate (40 µg/Lane)

Predicted band size: 48 kDa
Observed band size: 40-50 kDa

Exposure time: Lane 1-4: 11 seconds; Lane 5-6: 2 minutes 24 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 (HA600021) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
HA600021_2.jpg Fig2: Immunocytochemistry analysis of HeLa cells labeling Cytokeratin 18 with Mouse anti-Cytokeratin 18 antibody (HA600021) at 1/100 dilution.

Cells were fixed in 100% precooled methanol 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 Mouse anti-Cytokeratin 18 antibody (HA600021) 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.
HA600021_3.jpg Fig3: Immunohistochemical analysis of paraffin-embedded human liver tissue using anti-Cytokeratin 18 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA600021, 1/400) for 30 minutes 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.
HA600021_4.jpg Fig4: Immunohistochemical analysis of paraffin-embedded human skin tissue using anti-Cytokeratin 18 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA600021, 1/400) for 30 minutes 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.
HA600021_5.jpg Fig5: Immunohistochemical analysis of paraffin-embedded human prostate carcinoma tissue using anti-Cytokeratin 18 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA600021, 1/400) for 30 minutes 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.
HA600021_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human breast carcinoma tissue using anti-Cytokeratin 18 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 8.0-8.4) for 20 minutes.The tissues were blocked in 5% BSA for 30 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA600021, 1/400) for 30 minutes 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.
HA600021_7.jpg Fig7: Flow cytometric analysis of Cytokeratin 18 was done on MCF-7 cells. The cells were fixed, permeabilized and stained with the primary antibody (HA600021, 1/50) (red). After incubation of the primary antibody at room temperature for an hour, the cells were stained with a Alexa Fluor 488-conjugated Goat anti-Mouse IgG Secondary antibody at 1/1000 dilution for 30 minutes.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”.