Antibodies, Proteins, Kits and Reagents for Life Science

288 134 ₸ Product size

100 µl 288 134 ₸

Order now and get it on Tuesday March 02, 2021

Anti-CD14 antibody [EPR21847] (ab221678)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EPR21847] to CD14
Suitable for: IP, Flow Cyt, ICC/IF, WB
Reacts with: Mouse

Product name

Anti-CD14 antibody [EPR21847]
See all CD14 primary antibodies
Description

Rabbit monoclonal [EPR21847] to CD14
Host species

Rabbit

Tested Applications & Species

Application	Species
Flow Cyt	Mouse
ICC/IF	Mouse
IP	Mouse
WB	Mouse

See all applications and species data

Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: J774A.1 and RAW264.7 whole cell lysate; mouse lymph node and placenta lysates. ICC/IF: J774A.1 and RAW 264.7 cells. Flow cyt: RAW 264.7 cells. IP: RAW 264.7 whole cell lysate.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR21847
Isotype

IgG
Research areas

Flow Cytometry - Anti-CD14 antibody [EPR21847] (ab221678)

Flow cytometric analysis of RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cell line labeling CD14 with ab221678 at 1/500 dilution (red) compared with a Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells incubated with secondary antibody only) (blue). Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)(ab150077) at 1/2000 dilution was used as the secondary antibody.

Gated on viable cells.
Immunoprecipitation - Anti-CD14 antibody [EPR21847] (ab221678)

CD14 was immunoprecipitated from 0.35 mg RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate with ab221678 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab221678 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1: RAW 264.7 (Mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate 10 μg (Input).
Lane 2: ab221678 IP in RAW 264.7 whole cell lysate(+).
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab221678 in RAW 264.7 whole cell lysate (-).

Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 8 seconds

The molecular mass observed is consistent with the literature (PMID: 9502426; PMID:7513013)
Western blot - Anti-CD14 antibody [EPR21847] (ab221678)

All lanes : Anti-CD14 antibody [EPR21847] (ab221678) at 1/1000 dilution

Lane 1 : J774A.1 (mouse reticulum cell sarcoma monocyte macrophage), whole cell lysate at 10 µg
Lane 2 : Mouse lymph node lysate at 20 µg
Lane 3 : RAW264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage), whole cell lysate at 10 µg
Lane 4 : Mouse placenta lysate at 10 µg

Secondary
Lanes 1-3 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution
Lane 4 : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Developed using the ECL technique.

Predicted band size: 39 kDa
Observed band size: 50-55 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

Exposure time: Lane 1: 6 seconds; Lane 2: 3 minutes; Lane 3: 10 seconds; Lane 4: 81 seconds.

The molecular mass observed is consistent with the literature (PMID: 9502426; PMID: 7513013).
Immunocytochemistry/ Immunofluorescence - Anti-CD14 antibody [EPR21847] (ab221678)

Immunofluorescent analysis of 100% methanol-fixed RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) cells labeling CD14 with ab221678 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)(ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining on RAW 264.7 cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (re

Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488)(ab150077) secondary at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-CD14 antibody [EPR21847] (ab221678)

Immunofluorescent analysis of 100% methanol-fixed J774A.1 (mouse reticulum cell sarcoma monocyte macrophage) cells labeling CD14 with ab221678 at 1/1000 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous and cytoplasmic staining on J774A.1 cell line. The nuclear counter stain is DAPI (blue).

Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) (ab195889) at 1/200 dilution (red).

Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary at 1/1000 dilution.
Anti-CD14 antibody [EPR21847] (ab221678)

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