EM1901-32

Cytokeratin 14 Mouse Monoclonal Antibody [A2C11]

cat.: EM1901-32

Product Type:	Mouse monoclonal IgG1, primary antibodies
Species reactivity:	Human
Applications:	WB, IHC-P, IF-Cell
Clonality:	Monoclonal
Clone number:	A2C11

Form:	Liquid
Storage condition:	Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer:	1*PBS (pH7.4), 0.2% BSA, 50% Glycerol. Preservative: 0.05% Sodium Azide.
Concentration:	2ug/ul
Purification:	Protein G affinity purified.
Molecular weight:	Predicted band size: 52 kDa
Isotype:	IgG1
Immunogen:	Synthetic peptide within Human Cytokeratin 14 aa 423-472 / 472.
Positive control:	A431 cell lysate, A431, human lung squamous carcinoma tissue, human skin tissue, human tonsil tissue.
Subcellular location:	Cytoplasm, Nucleus.
Recommended Dilutions: WB IHC-P IF-Cell	1:500-1:2,000 1:1:1,000 1:100
Uniprot #:	SwissProt: P02533 Human
Alternative names:	CK 14 CK-14 ck14 Cytokeratin 14 Cytokeratin-14 Cytokeratin14 Dowling Meara EBS3 EBS4 Epidermolysis bullosa simplex K14 K1C14_HUMAN Keratin 14 (epidermolysis bullosa simplex, Dowling-Meara, Koebner) Keratin 14 Keratin Keratin type I cytoskeletal 14 Keratin, type I cytoskeletal 14 Keratin-14 Keratin14 Koebner Krt 14 Krt14 NFJ OTTHUMP00000164624 type I cytoskeletal 14

Images

Fig1: Western blot analysis of Cytokeratin 14 on different lysates with Mouse anti-Cytokeratin 14 antibody (EM1901-32) at 1/1,000 dilution.

Lane 1: A431 cell lysate
Lane 2: MCF7 cell lysate (negative)
Lane 3: PANC-1 cell lysate (negative)

Lysates/proteins at 20 µg/Lane.

Predicted band size: 52 kDa
Observed band size: 52 kDa

Exposure time: 1 minute 59 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 (EM1901-32) 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.

Fig2: Immunocytochemistry analysis of A431 (positive) and PANC-1 (negative) labeling Cytokeratin 14 with Rabbit anti-Cytokeratin 14 antibody (EM1901-32) 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-Cytokeratin 14 antibody (EM1901-32) 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 lung squamous carcinoma tissue with Mouse anti-Cytokeratin 14 antibody (EM1901-32) at 1/1,000 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 (EM1901-32) 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 skin tissue with Mouse anti-Cytokeratin 14 antibody (EM1901-32) at 1/1,000 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 (EM1901-32) 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 tonsil tissue with Mouse anti-Cytokeratin 14 antibody (EM1901-32) at 1/1,000 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 (EM1901-32) 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.

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