CD14 Recombinant Rabbit Monoclonal Antibody [SC69-02]
cat.: HA750232
Product Type: Recombinant Rabbit monoclonal IgG, primary antibodies
Species reactivity: Human, Mouse
Applications: WB, IF-Cell, IHC-P, FC
Clonality: Monoclonal
Clone number: SC69-02
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*PBS (pH7.4).
Concentration: 1ug/ul
Purification: Protein A affinity purified.
Molecular weight: Predicted band size: 40 kDa
Isotype: IgG
Immunogen: Synthetic peptide within Human CD14 aa 310-335 / 375.
Positive control: THP-1 cell lysate, RAW264.7 cell lysate, NIH/3T3 cell lysates, A549, NCCIT, NIH/3T3, LO2, human tonsil tissue, human liver tissue, human colon carcinoma tissue, human spleen tissue, human uterus tissue, human lymph nodes tissue, human cervical cancer.
Subcellular location: Cell membrane, Secreted, Golgi apparatus, Membrane raft.
Recommended Dilutions:
  WB
  IF-Cell
  IHC-P
  FC
1:5,000
1:50-1:200
1:200-1:800
1:1,000
Uniprot #: SwissProt: P08571 Human | P10810 Mouse
Alternative names: CD 14 CD_antigen=CD14 CD14 CD14 antigen CD14 molecule CD14_HUMAN LPS-R Mo2 Monocyte differentiation antigen CD14 Monocyte differentiation antigen CD14 urinary form Monocyte differentiation antigen CD14, membrane-bound form Myeloid cell specific leucine rich glycoprotein Myeloid cell-specific leucine-rich glycoprotein
Images
HA750232_1.jpg Fig1: Western blot analysis of CD14 on different lysates with Rabbit anti-CD14 antibody (HA750232) at 1/5,000 dilution.

Lane 1: THP-1 cell lysate
Lane 2: RAW264.7 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 40 kDa
Observed band size: 60 kDa

Exposure time: 1 minute 40 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 (HA750232) at 1/5,000 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.
HA750232_2.jpg Fig2: Western blot analysis of CD14 on different lysates with Rabbit anti-CD14 antibody (HA750232) at 1/1,000 dilution.

Lane 1: SW480-si NT cell lysate
Lane 2: SW480-si CD14 cell lysate

Lysates/proteins at 10 µg/Lane.

Predicted band size: 40 kDa
Observed band size: 50-60 kDa

Exposure time: 7 seconds; ECL: merk
4-20% SDS-PAGE gel.

ET1610-85 was shown to specifically react with CD14 in SW480-si NT cells. Weakened band was observed when SW480-si CD14 sample was tested. SW480-si NT and SW480-si CD14 samples were subjected to SDS-PAGE. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM in TBST for 1 hour at room temperature. The primary antibody (ET1610-85, 1/1,000) and Loading control antibody (Rabbit anti-GAPDH, ET1601-4, 1/10,000) were used in 5% BSA at room temperature for 2 hours. Goat Anti-rabbit IgG-HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
HA750232_3.jpg Fig3: Immunocytochemistry analysis of THP-1 cells labeling CD14 with Rabbit anti-CD14 antibody (HA750232) 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-CD14 antibody (HA750232) 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.
HA750232_4.jpg Fig4: Immunocytochemistry analysis of NIH/3T3 cells labeling CD14 with Rabbit anti-CD14 antibody (HA750232) 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-CD14 antibody (HA750232) 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.
HA750232_5.jpg Fig5: Flow cytometric analysis of human peripheral blood cells labelling CD14 (HA750232).
HA750232_6.jpg Fig6: Immunohistochemical analysis of paraffin-embedded human liver tissue with Rabbit anti-CD14 antibody (HA750232) at 1/800 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 (HA750232) at 1/800 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.
HA750232_7.jpg Fig7: Immunohistochemical analysis of paraffin-embedded human lymph nodes tissue with Rabbit anti-CD14 antibody (HA750232) at 1/800 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 (HA750232) at 1/800 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.
HA750232_8.jpg Fig8: Immunohistochemical analysis of paraffin-embedded human spleen tissue with Rabbit anti-CD14 antibody (HA750232) at 1/200 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 (HA750232) at 1/200 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”.