Immunocytochemistry/ Immunofluorescence: CD14 Antibody (M5E2) [NB100-77758] - Representative immunostaining of CD14 (red) in adult human lung tissue sample with nuclei counterstained blue. This image was submitted by ...read more
Flow Cytometry: CD14 Antibody (M5E2) [NB100-77758] - A surface stain was performed on human peripheral blood monocytes with CD163 (EDHu-1) antibody NB110-40686APC and a matched isotype control NBP2-27287APC. Cells were ...read more
Immunocytochemistry/ Immunofluorescence: CD14 Antibody (M5E2) [NB100-77758] - Human lung tissue was stained with FITC-conjugated CD14 (Green) for overnight. Image from the Alexa Fluor 488 version of this antibody.
Flow Cytometry: CD14 Antibody (M5E2) [NB100-77758] - Human peripheral blood monocytes stained with M5E2 APC
Flow Cytometry: CD14 Antibody (M5E2) [NB100-77758] - Analysis using the FITC conjugate of NB100-77758. Staining of Human peripheral blood monocytes with M5E2 FITC.
Flow Cytometry: CD14 Antibody (M5E2) [NB100-77758] - Analysis using the PE conjugate of NB100-77758. Staining of CD14 in human PBMC using anti-CD14 antibody conjugated with PE. Image from verified customer review.
Flow Cytometry: CD14 Antibody (M5E2) [NB100-77758] - Analysis using PE conjugate of NB100-77758. A cell surface stain was performed on hPBMCs with ILT5 antibody (MM0413-9S32) NBP2-11729 (top image) and a matched isotype ...read more
Dual RNAscope ISH-IHC: CD14 Antibody (M5E2) [NB100-77758] - TNFA mRNA (red) and CD14 protein (green) were detected in formalin-fixed paraffin-embedded tissue sections of human malignant lymph node. ACD's Integrated ...read more
The M5E2 antibody inhibits monocyte activation and cytokine production induced by LPS. Additional reported applications (for the relevant formats) include: immunohistochemical staining of acetone-fixed frozen sections, blocking of LPS stimulation, and immunofluorescence microscopy. Clone M5E2 is not recommended for immunohistochemical staining of formalin-fixed paraffin-embedded sections. This antibody is CyTOF ready. See Simple Western Antibody Database for Simple Western validation: tested in THP-1; antibody dilution of 50 ug/ml; separated by size; detects a band at 54 kDa
Expected to cross react with Chimpanzee, Cynomolgus, Rhesus, Pigtailed Macaque, Capuchin Monkey, Cotton-topped Tamarin, Common Marmoset, Squirrel Monkey.
Packaging, Storage & Formulations
Storage
Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.
Buffer
PBS
Preservative
0.02% Sodium Azide
Concentration
1.0 mg/ml
Purity
Protein G purified
Alternate Names for CD14 Antibody (M5E2) - BSA Free
CD14 antigen
CD14 molecule
CD14
monocyte differentiation antigen CD14
Myeloid cell-specific leucine-rich glycoprotein
Background
CD14 is a 53-55 kD glycosylphosphatidylinositol (GPI)-linked membrane glycoprotein also known as LPS receptor. CD14 is expressed at high levels on monocytes and macrophages, and at lower levels on granulocytes. Some dendritic cell populations such as interfollicular dendritic cells, reticular dendritic cells, and Langerhans cells have also been reported to express CD14. As a high-affinity receptor for LPS, CD14 is involved in the clearance of gram-negative pathogens and in the upregulation of adhesion molecules and cytokines expression in monocytes and neutrophils. The M5E2 antibody inhibits monocyte activation and cytokine production induced by LPS.
Limitations
This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.
*Low Rating Note: Novus Innovators Program - new species or application used on a primary antibody. Dewaxing to water: 1, paraffin sections in turn sliced into 15 min - dimethylbenzene xylene Ⅰ Ⅱ 15 min - xylene III Ⅰ 5 min - 15 min - anhydrous ethanol anhydrous ethanol Ⅱ 5 min 5 min - 75% - 85% alcohol alcohol 5 min - the distilled water to wash. 2, antigen repair: biopsy under full antigen repair citrate buffer (PH6.0) repair box in a have a certain amount of water pressure cooker, induction cooker heated to stomatal began to jet, stop heating, release the pressure, the slice on the repair box, induction cooker heated to stomatal began to jet, 5 min after close the induction cooker, the process should prevent excessive evaporation buffer, do not dry.After natural cooling, the slides were placed in PBS (PH7.4) and shaken on the decolorizing shaper for 3 times, 5min each. 3. Break endogenous peroxidase: the slides were placed in 3% hydrogen peroxide solution and incubated at room temperature and away from light for 25 min. The slides were placed in PBS (PH7.4) and shaken on a decolorizing shaper for 3 times for 5min each. Serum sealing: 3%BSA was added to cover the tissue evenly in the chemochemical ring, and the tissue was sealed at room temperature for 30min.(Primary antibody is sealed with rabbit serum from goat and BSA from other sources.) 4. Primary antibody was added: the blocking solution was gently removed, the primary antibody prepared with PBS in a certain proportion was added to the sections, and the sections were placed flat in a wet box and incubated overnight at 4°C.(Add a small amount of water in the wet box to prevent evaporation of antibodies) 5. Secondary antibody was added: the slides were placed in PBS (PH7.4) and washed by shaking on the decolorizing shaker for 3 times, 5min each.After the sections were slightly shaken and dried, the tissues were dripped with secondary antibody (HRP-labeled) of the species corresponding to the primary antibody and incubated at room temperature for 50min. 7. DAB color development: The slides were placed in PBS (PH7.4) and shaken on the decoloring shaker for 3 times, 5min each.DAB color developing solution newly prepared was added in the circle after the sections were slightly dried. The color developing time was controlled under the microscope. The positive color was brownish yellow, and the color developing was stopped after the sections were washed under tap water. 8. Restaining nuclei: restaining with hematoxylin for about 3min; washing with tap water; differentiation with hematoxylin differentiation solution for several seconds; washing with tap water; flushing with hematoxylin returning blue solution; washing with running water. , dehydration and 9: the slice in turn into 5 min 5 min 75% alcohol - 85% alcohol, anhydrous ethanol Ⅰ Ⅱ 5 min - the anhydrous ethanol dehydration in 5 min - xylene Ⅰ 5 min is transparent, the slice out of xylene is a bit dry, neutral rubber sealing piece. Microscope examination, image acquisition and analysis.
The whole blood cells were lysed to get rid of the RBC. The washed cell were then incubated with the Antibody. They were washed and then incubated with a conjugated secondary. Allowed to sit for 20mins and then washed again and read on the BD Fortessa.
Product General Protocols
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FAQs for CD14 Antibody (NB100-77758). (Showing 1 - 2 of 2 FAQs).
We have an inquiry of your product Cat. ID # NB100-2735, NB600-1185, NB100-77758, NB100-78016, NB100-77855. We are interested in this product and would like to make sure that there is no any other protein added in the buffer. I saw the storage buffer in datasheet doesn't has any other protein but still would like to confirm with you again. Would you please help us to check and provide the purification method and buffer of the antibody?
Thank you for your inquiry. Catalog numbers NB100-77758, NB100-78016 and NB100-77855 are in a buffer containing PBS and sodium azide. NB600-1185 is in a buffer containing PBS, 150mM NaCl and sodium azide. NB100-2735 is whole tissue culture supernatant and may therefore contain other proteins.
Can this antibody be used for testing frozen and not fixed human (CNS, brain) tissue?
The CD14 Antibody (M5E2) antibody with catalog number NB100-77758 is covered by the Novus Guarantee to detect human protein in frozen samples by IHC. The recommended dilution range for IHC-Frozen is 1:10-1:2000. According to UniProt, CD14 is expressed strongly on the surface of monocytes, weakly on the surface of granulocytes, and is also expressed by most tissue macrophages. The following PubMed reference discusses the expression and release of CD14 in astrocytic brain tumors.
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CD14 - TLR4 is my friend in battle against infections! CD14 is a well-characterized cell-activating receptor for lipopolysaccharide-binding protein (LBP) and peptidoglycan. It is an important modulator for lipopolysaccharide (LPS)-dependent signaling and is a component of the multi-protein complex contain... Read full blog post.
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