Anti-PTBP1 antibody [EPR9049(B)]

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-PTBP1 antibody [EPR9049(B)] (AB134950)

Immunofluorescent analysis of HeLa cells labelling PTBP1 with ab134950 at 1/100 dilution.

• WB

Lab

Western blot - Anti-PTBP1 antibody [EPR9049(B)] (AB134950)

Lanes 1 - 2 : Merged signal (red and green). Green - ab134950 observed at 57 kDa. Red - loading control, ab9484, observed at 37 kDa.

ab134950 was shown to specifically react with PTBP1 in wild-type HAP1 cells as signal was lost in PTBP1 knockout cells. Wild-type and PTBP1 knockout samples were subjected to SDS-PAGE. ab134950 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed ab216776 secondary antibodies at 1/10000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PTBP1 antibody [EPR9049(B)] (ab134950) at 1/1000 dilution

Lane 1:

Wild-type HAP1 whole cell lysate at 20 µg

Lane 2:

PTBP1 knockout HAP1 whole cell lysate at 20 µg

Predicted band size: 57 kDa

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• WB

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Western blot - Anti-PTBP1 antibody [EPR9049(B)] (AB134950)

All lanes:

Western blot - Anti-PTBP1 antibody [EPR9049(B)] (ab134950) at 1/1000 dilution

Lane 1:

Daudi cell lysate at 10 µg

Lane 2:

HeLa cell lysate at 10 µg

Lane 3:

Jurkat cell lysate at 10 µg

Lane 4:

BXPC3 cell lysate at 10 µg

Secondary

All lanes:

HRP labelled goat anti-rabbit at 1/2000 dilution

Predicted band size: 57 kDa

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• WB

Lab

Western blot - Anti-PTBP1 antibody [EPR9049(B)] (AB134950)

Lanes 1- 2 : Merged signal (red and green). Green - ab134950 observed at 57 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.

ab134950 was shown to react with PTBP1 in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line ab265155 (knockout cell lysate ab257614) was used. Wild-type HeLa and PTBP1 knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab134950 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-PTBP1 antibody [EPR9049(B)] (ab134950) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

PTBP1 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human PTBP1 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-ptbp1-knockout-hela-cell-line-ab265155'>ab265155</a>)

Predicted band size: 57 kDa

Observed band size: 57 kDa

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• OI-RD Scanning

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OI-RD Scanning - Anti-PTBP1 antibody [EPR9049(B)] (AB134950)

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.