Anti-CD14 antibody [EPR21847] - BSA and Azide free

• IHC-Fr

Lab

Immunohistochemistry (Frozen sections) - Anti-CD14 antibody [EPR21847] - BSA and Azide free (AB231852)

This data was developed using ab221678 ,the same antibody clone in a different buffer formulation.

Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse spleen tissue labeling CD14 with ab221678 at 1/50 (10.62 ug/ml) dilution followed by ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor ® 488) at 1/1000 dilution (Green). Positive staining on mouse spleen. is observed. The nuclear counterstain was DAPI (Blue).

Secondary antibody control : Secondary antibody is ab150077 Goat Anti-Rabbit IgG H&L (Alexa Fluor ®; 488) at 1000 dilution.

Heat mediated antigen retrieval using sodium citrate buffer (10mM citrate pH 6.0 + 0.05% Tween-20).

• Flow Cyt

Lab

Flow Cytometry - Anti-CD14 antibody [EPR21847] - BSA and Azide free (AB231852)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab221678).

Flow cytometry staining of C57 BL/6 mouse bone marrow cells with ab221678 (right) or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (left). Cells were incubated for 30 min on ice in 1x PBS containing 10μg/ml anti CD16/CD32 and 10% normal goat serum to block FC receptors and non-specific protein-protein interaction followed by the antibody ab221678 or Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (1x 106 in 100 μl at 10.0 μg/ml (1/215)) for 30min on ice. The cells were simultaneously stained with Ly6G.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min on ice

Acquisition of >30000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter. Events were gated on viable cells.

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD14 antibody [EPR21847] - BSA and Azide free (AB231852)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab221678).

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD14 antibody [EPR21847] - BSA and Azide free (AB231852)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab221678).

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-CD14 antibody [EPR21847] - BSA and Azide free (AB231852)

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.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab221678).

• IP

Supplier Data

Immunoprecipitation - Anti-CD14 antibody [EPR21847] - BSA and Azide free (AB231852)

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)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab221678).

All lanes:

Immunoprecipitation - Anti-CD14 antibody [EPR21847] (<a href='/en-us/products/primary-antibodies/cd14-antibody-epr21847-ab221678'>ab221678</a>)

Predicted band size: 40 kDa

Observed band size: 50-55 kDa

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