Anti-DCTN3 antibody [EPR5097]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-DCTN3 antibody [EPR5097] (AB124674)

Immunofluorescence staining of BxPC-3 cells with purified ab124674 at a working dilution of 1 in 100, counter-stained with DAPI. The secondary antibody was Alexa Fluor^® 488 goat anti rabbit (ab150077), used at a dilution of 1 in 500. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative control is shown in bottom right hand panel - for the negative control, purified ab124674 was used at a dilution of 1/200 followed by an Alexa Fluor^® 594 goat anti-mouse antibody (ab15120) at a dilution of 1/500.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DCTN3 antibody [EPR5097] (AB124674)

Immunohistochemical staining of DCTN3 in paraffin embedded human breast tissue by unpurified ab124674 at 1/100 dilution.

Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-DCTN3 antibody [EPR5097] (AB124674)

Immunohistochemical staining of paraffin embedded human pancreas with purified ab124674 at a working dilution of 1 in 100. The secondary antibody used is a HRP polymer for rabbit IgG. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.

• WB

Lab

Western blot - Anti-DCTN3 antibody [EPR5097] (AB124674)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-DCTN3 antibody [EPR5097] (ab124674) at 1/10000 dilution

Lane 1:

293T cell lysate at 10 µg

Lane 2:

SH-SY5Y cell lysate at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

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• WB

Unknown

Western blot - Anti-DCTN3 antibody [EPR5097] (AB124674)

All lanes:

Western blot - Anti-DCTN3 antibody [EPR5097] (ab124674) at 1/1000 dilution

Lane 1:

293T cell lysates at 10 µg

Lane 2:

Mouse muscle lysates at 10 µg

Lane 3:

HeLa cell lysates at 10 µg

Lane 4:

BxPC 3 cell lysates at 10 µg

Lane 5:

SH-SY5Y cell lysates at 10 µg

Lane 6:

Rat muscle lysates at 10 µg

Secondary

All lanes:

Goat anti-Rabbit HRP at 1/2000 dilution

Predicted band size: 21 kDa

false

• WB

Lab

Western blot - Anti-DCTN3 antibody [EPR5097] (AB124674)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-DCTN3 antibody [EPR5097] (ab124674) at 1/2000 dilution

All lanes:

Mouse muscle at 10 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

false

• WB

Lab

Western blot - Anti-DCTN3 antibody [EPR5097] (AB124674)

Blocking buffer : 5% NFDM/TBST

Dilution buffer : 5% NFDM/TBST

All lanes:

Western blot - Anti-DCTN3 antibody [EPR5097] (ab124674) at 1/10000 dilution

All lanes:

Rat muscle at 20 µg

Secondary

All lanes:

HRP goat anti-rabbit (H+L) at 1/1000 dilution

Predicted band size: 21 kDa

Observed band size: 21 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-DCTN3 antibody [EPR5097] (AB124674)

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.