Anti-JunB antibody [EPR6518]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-JunB antibody [EPR6518] (AB128878)

Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling JunB with purified ab128878 at 1 : 200 dilution (9.1 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) 1 : 200 (2.5 μg/ml). Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) was used as the secondary antibody at 1 : 1000 (2 μg/ml) dilution. DAPI (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-JunB antibody [EPR6518] (AB128878)

Unpurified ab128878, at a 1/250 dilution, staining JunB in MCF7 (Human breast adenocarcinoma cell line) cells by Immunofluorescence.

• IP

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Immunoprecipitation - Anti-JunB antibody [EPR6518] (AB128878)

ab128878 (purified) at 1 : 100 dilution (2 μg) immunoprecipitating JunB in HeLa whole cell lysates.

Lane 1 (input) : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates 10ug
Lane 2 (+) : ab128878 & HeLa whole cell lysates
Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab128878 in HeLa whole cell lysate

For western blotting, VeriBlot for IP Detection Reagent (HRP) (ab131366) was used at 1 : 1000 dilution.

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-JunB antibody [EPR6518] (ab128878)

Predicted band size: 112 kDa,31 kDa,36 kDa,38 kDa,40 kDa,55 kDa

Observed band size: 45 kDa

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• WB

Lab

Western blot - Anti-JunB antibody [EPR6518] (AB128878)

All lanes:

Western blot - Anti-JunB antibody [EPR6518] (ab128878) at 1/1000 dilution

Lane 1:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Lane 2:

Mouse uterus lysates at 15 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: 36 kDa

Observed band size: 43 kDa

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• WB

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Western blot - Anti-JunB antibody [EPR6518] (AB128878)

All lanes:

Western blot - Anti-JunB antibody [EPR6518] (ab128878) at 1/1000 dilution

Lane 1:

MCF-7 lysate at 10 µg

Lane 2:

Human tonsil lysate at 10 µg

Lane 3:

Human uterus cancer lysate at 10 µg

Lane 4:

HeLa lysate at 10 µg

Lane 5:

HACAT lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 36 kDa

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• WB

Lab

Western blot - Anti-JunB antibody [EPR6518] (AB128878)

All lanes:

Western blot - Anti-JunB antibody [EPR6518] (ab128878) at 1/200 dilution

Lane 1:

Rat lung lysates at 15 µg

Lane 2:

Rat spleen lysates at 15 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: 36 kDa

Observed band size: 43 kDa

false

• WB

Lab

Western blot - Anti-JunB antibody [EPR6518] (AB128878)

All lanes:

Western blot - Anti-JunB antibody [EPR6518] (ab128878) at 1/1000 dilution

All lanes:

Human uterus lysates at 15 µg

Secondary

All lanes:

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

Predicted band size: 36 kDa

Observed band size: 43 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-JunB antibody [EPR6518] (AB128878)

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.