Anti-CD16 antibody [EPR22409-124] - BSA and Azide free

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD16 antibody [EPR22409-124] - BSA and Azide free (AB252908)

Immunohistochemical analysis of paraffin-embedded human spleen tissue labeling CD16 with ab246222 at 1/500 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on red pulp of human spleen (PMID : 26512139; 29692344). Counter stained with hematoxylin.

Secondary antibody only control : Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).

Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 minutes.

The section was incubated with ab246222 for 10 minutes at 37°C. The immunostaining staining was performed on a Leica Biosystems BOND^® RX instrument.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab246222).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-CD16 antibody [EPR22409-124] - BSA and Azide free (AB252908)

Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized human PBMCs (human peripheral blood mononuclear cells) or Raji (human Burkitt's lymphoma cell line) cell line labeling CD16 with ab246222 at 1/100 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in subsets of human PBMC cells. 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 antibody at 1/1000 dilution.

Negative control : Raji (PMID : 11207281).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab246222).

• Flow Cyt

Supplier Data

Flow Cytometry - Anti-CD16 antibody [EPR22409-124] - BSA and Azide free (AB252908)

Negative control : Raji. (PMID : 11207281).

Raji (human Burkitt's lymphoma cell line) were stained with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)(Left) or ab246222 (Right) followed by a Goat anti rabbit IgG (Dylight ^® 488) at 1/2000 dilution.

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 (ab246222).

• Flow Cyt

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Flow Cytometry - Anti-CD16 antibody [EPR22409-124] - BSA and Azide free (AB252908)

Human PBMCs (human peripheral blood mononuclear cells) were stained with Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)(Left) or ab246222 (Right) followed by a Goat anti rabbit IgG (Dylight ^® 488) at 1/2000 dilution. They were then stained with Alexa Fluor^® 647-conjugated anti-CD14. The expression pattern is consistent with what described in the literature. (PMID : 21738687)

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 (ab246222).

• IP

Unknown

Immunoprecipitation - Anti-CD16 antibody [EPR22409-124] - BSA and Azide free (AB252908)

CD16 was immunoprecipitated from 0.35 mg of human spleen lysate with ab246222 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab246222 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/5000 dilution.

Lane 1 : Human spleen lysate 10 μg (Input).

Lane 2 : ab246222 IP in human spleen lysate.

Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab246222 in human spleen lysate.

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

Exposure time : 5 seconds.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab246222).

All lanes:

Immunoprecipitation - Anti-CD16 antibody [EPR22409-124] (<a href='/en-us/products/primary-antibodies/cd16-antibody-epr22409-124-ab246222'>ab246222</a>)

Predicted band size: 29 kDa

Observed band size: 50-70 kDa

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