Anti-IGF2BP2/IMP-2 antibody [EPR6741(B)]

• IHC-P

Unknown

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IGF2BP2/IMP-2 antibody [EPR6741(B)] (AB124930)

ab124930 (unpurified), at 1/50 dilution, staining IGF2BP2/IMP-2 in formalin-fixed, paraffin-embedded Human colon tissue by immunohistochemistry.

Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-IGF2BP2/IMP-2 antibody [EPR6741(B)] (AB124930)

Immunocytochemistry/ Immunofluorescence analysis of HepG2 (Human liver hepatocellular carcinoma cell line) cells labeling IGF2BP2/IMP-2 with purified ab124930 at 1/250 dilution (3.8μg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with ab195889, an anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at 1/200 (2.5 μg/ml). ab150077, a Goat anti-rabbit IgG (Alexa Fluor^® 488) was used as the secondary antibody at 1/1000 dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-IGF2BP2/IMP-2 antibody [EPR6741(B)] (AB124930)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung carcinoma tissue sections labeling IGF2BP2/IMP-2 with purified ab124930 at 1/200 dilution (4.8 μg/ml). Heat mediated antigen retrieval was performed using EDTA Buffer, PH9. Hematoxylin was used to counter stain. ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody at 1/0 dilution. PBS instead of the primary antibody was used as the negative control.

• WB

Supplier Data

Western blot - Anti-IGF2BP2/IMP-2 antibody [EPR6741(B)] (AB124930)

Blocking and diluting buffer : 5% NFDM/TBST.

ab205606 was used as a loading control at 1/1000 dilution.

ab179807 was used at 1/1000 dilution.

Lanes 1 - 3:

Western blot - Anti-IGF2BP2/IMP-2 antibody [EPR6741(B)] (ab124930) at 1/1000 dilution

Lanes 1 - 3:

Western blot - Anti-IMP3 antibody [EPR12021-114] (<a href='/en-us/products/primary-antibodies/imp3-antibody-epr12021-114-ab179807'>ab179807</a>) at 1/1000 dilution

Lane 1:

C-Myc/DDK-tagged full length Human IGF2BP1 recombinant protein at 0.01 µg

Lane 2:

C-Myc/DDK-tagged full length Human IGF2BP2 recombinant protein at 0.01 µg

Lane 3:

N-His-tagged full length Human IGF2BP3 recombinant protein at 0.01 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: 66 kDa

Observed band size: 75 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-IGF2BP2/IMP-2 antibody [EPR6741(B)] (AB124930)

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

The double bands are consistent with what has been described in PMID 23177649.

All lanes:

Western blot - Anti-IGF2BP2/IMP-2 antibody [EPR6741(B)] (ab124930) at 1/5000 dilution

Lane 1:

293T (Human embryonic kidney epithelial cell) whole cell lysates at 20 µg

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 3:

HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysates at 20 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: 66 kDa

Observed band size: 60 kDa,62 kDa

false

Exposure time: 60s

• WB

Lab

Western blot - Anti-IGF2BP2/IMP-2 antibody [EPR6741(B)] (AB124930)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/1000000 dilution.

All lanes:

Western blot - Anti-IGF2BP2/IMP-2 antibody [EPR6741(B)] (ab124930) at 1/1000 dilution

Lane 1:

Human adrenal gland tissue lysate at 20 µg

Lane 2:

Mouse adrenal gland tissue lysate 20ug at 20 µg

Lane 3:

Mouse brain tissue lysate at 20 µg

Lane 4:

Mouse heart tissue lysate at 20 µg

Secondary

All lanes:

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

Observed band size: 70 kDa,15 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-IGF2BP2/IMP-2 antibody [EPR6741(B)] (AB124930)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-IGF2BP2/IMP-2 antibody [EPR6741(B)] (ab124930) at 1/2000 dilution

Lane 1:

293T (Human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 2:

SH-SY5Y (Human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Rat brain lysate at 20 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: 66 kDa

Observed band size: 66 kDa

false

• WB

Lab

Western blot - Anti-IGF2BP2/IMP-2 antibody [EPR6741(B)] (AB124930)

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-IGF2BP2/IMP-2 antibody [EPR6741(B)] (ab124930) at 1/10000 dilution

All lanes:

Mouse heart lysate 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: 66 kDa

Observed band size: 66 kDa

false

• OI-RD Scanning

Unknown

OI-RD Scanning - Anti-IGF2BP2/IMP-2 antibody [EPR6741(B)] (AB124930)

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.