Anti-CD14 antibody [EPR3653]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [EPR3653] (AB133335)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human colon tissue sections labeling CD14 with purified ab133335 at 1/2000 dilution (0.04 μg/ml). Heat mediated antigen retrieval was performed using ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control : PBS instead of the primary antibody. Hematoxylin was used as a counterstain.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [EPR3653] (AB133335)

Immunohistochemical analysis of Formalin-fixed, paraffin- embedded Human tonsil tissue labelling CD14 with ab133335 (unpurified) at 1/500 dilution. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [EPR3653] (AB133335)

Tissue Microarrays stained for "Anti-CD14 antibody [EPR3653]" using "ab133335"in immunohistochemical analysis. This table provides a detailed overview of positive (tick mark) and negative (cross mark) staining per sample type tested. The sections were pre-treated using Heat mediated antigen retrieval using Bond™ Epitope Retrieval Solution 2 (pH 9.0) for 20 minutes. The sections were incubated with ab133335 for 30 mins at room temperature followed by a ready to use Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). The immunostaining was performed on a Leica Biosystems BOND® RX instrument.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [EPR3653] (AB133335)

Immunohistochemical analysis of Formalin-fixed, paraffin- embedded Human placenta tissue labelling CD14 with ab133335 (unpurified) at 1/500 dilution. Heat mediated antigen retrieval was performed before commencing with IHC staining protocol.

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-CD14 antibody [EPR3653] (AB133335)

Immunohistochemical analysis of Formalin-fixed, paraffin- embedded human tonsil tissue labelling CD14 with ab133335 (unpurified) at 1/500 dilution. No blocking step performed. Anti-Rabbit HRP polymer was used as the secondary detection system. Heat-mediated antigen retrieval was performed using EDTA based pH 9.0 buffer.

• WB

Lab

Western blot - Anti-CD14 antibody [EPR3653] (AB133335)

Blocking/Diluting buffer and concentration : 5% NFDM/TBST

All lanes:

Western blot - Anti-CD14 antibody [EPR3653] (ab133335) at 1/1000 dilution

Lane 1:

Human tonsil lysates prepared in RIPA lysis method at 20 µg

Lane 2:

Human tonsil lysates prepared in 1%SDS Hot lysis method at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Predicted band size: 40 kDa

false

Exposure time: 45s

• WB

Lab

Western blot - Anti-CD14 antibody [EPR3653] (AB133335)

Blocking and diluting buffer : 5% NFDM/TBST

The expression level in HeLa and U-937 are low (PMID : 9886426 and 15730927)

All lanes:

Western blot - Anti-CD14 antibody [EPR3653] (ab133335) at 1/1000 dilution

Lane 1:

Human tonsil tissue lysate prepared in 1% SDS Hot lysis method at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma) whole cell lysate at 20 µg

Lane 3:

U-937 (Human histiocytic lymphoma) whole cell lysate at 20 µg

Secondary

Lane 1:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Lanes 2 - 3:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 40 kDa

Observed band size: 53 kDa

false

Exposure time: 3min

• WB

Unknown

Western blot - Anti-CD14 antibody [EPR3653] (AB133335)

All lanes:

Western blot - Anti-CD14 antibody [EPR3653] (ab133335) at 1/1000 dilution

Lane 1:

PBMC cell lysate prepared in 1% SDS Hot lysis method at 10 µg

Lane 2:

Human tonsil tissue lysate prepared in 1%SDS Hot lysis method at 10 µg

Lane 3:

SW480 cell lysate prepared in 1%SDS Hot lysis method at 10 µg

Secondary

All lanes:

Goat Anti-rabbit HRP at 1/2000 dilution

Predicted band size: 40 kDa

Observed band size: 53 kDa

false

• WB

Lab

Western blot - Anti-CD14 antibody [EPR3653] (AB133335)

Blocking and diluting buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-CD14 antibody [EPR3653] (ab133335) at 1/1000 dilution

All lanes:

His-Tagged Human CD14 (aa20 to 345) recombinant protein at 0.015 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 40 kDa

Observed band size: 43 kDa

false

Exposure time: 1s

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-CD14 antibody [EPR3653] (AB133335)

We have systematically measured KD (the equilibrium dissociation constant between the antibody and its antigen), of more than 840 recombinant antibodies to assess not only their individual KD values but also to see the average affinity of antibody. Based on the comparison with published literature values for mouse monoclonal antibodies, Recombinant antibodies appear to be on average 1-2 order of magnitude higher affinity.