Goat Anti-Mouse IgG H&L (HRP) preadsorbed

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

All lanes:

Western blot - Anti-hnRNP A1 antibody [9H10] (<a href='/en-us/products/primary-antibodies/hnrnp-a1-antibody-9h10-ab5832'>ab5832</a>) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

Lane 3:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 4:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 38 kDa

Observed band size: 37 kDa,43 kDa

true

Exposure time: 30s

• IP

Lab

Immunoprecipitation - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

Lane 1 : HEK293 cell lysate overexpressing GFP - 10ug

Lane 2 : Unbound IP supernatant : Anti-GFP antibody (Magnetic beads) + 100ug HEK293 cell lysate overexpressing GFP

Lane 3 : Negative IP : off-target sdAb beads : Anti-GFP antibody (Magnetic beads) + 100ug HEK293 cell lysate overexpressing GFP

Lane 4 : IP : Anti-GFP antibody (Magnetic beads) + 100ug HEK293 cell lysate overexpressing GFP

Lane 5 : HEK293 cell lysate overexpressing GFP - 10ug

Lane 6 : Unbound IP supernatant : anti-GFP antibody (ab290) + Protein A agarose + 100ug HEK293 cell lysate overexpressing GFP

Lane 7 : IP : anti-GFP antibody (ab290) + Protein A agarose+ 100ug HEK293 cell lysate overexpressing GFP

Anti-GFP antibody (Magnetic beads) (ab193983) was used to pull down GFP from HEK293 cell lysate overexpressing GFP. ab290 (Rabbit polyclonal to GFP - ChIP Grade) was used as a positive control. The gel was stained with Optiblot Blue (ab119211).

All lanes:

Immunoprecipitation - Magnetic beads Anti-GFP VHH Single Domain antibody (<a href='/en-us/products/primary-antibodies/magnetic-beads-gfp-vhh-single-domain-antibody-ab193983'>ab193983</a>) at 1/500 dilution

Secondary

All lanes:

Immunoprecipitation - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/20000 dilution

Observed band size: 48 kDa

false

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

All lanes:

Western blot - Anti-ABCA4 antibody [3F4] (<a href='/en-us/products/primary-antibodies/abca4-antibody-3f4-ab77285'>ab77285</a>) at 1/500 dilution

Lane 1:

WERI (Human Retinoblastoma) Whole Cell Lysate at 10 µg

Lane 2:

Y79 (Human retinoblastoma cell line) Whole Cell Lysate at 10 µg

Lane 3:

Rat Retina Tissue Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 256 kDa

Observed band size: 142 kDa,257 kDa,51 kDa

true

Exposure time: 20min

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab7291 overnight at 4°C. Antibody binding was detected using an anti-mouse HRP (ab97040), and visualised using ECL development solution ab133406

All lanes:

Western blot - Anti-alpha Tubulin antibody [DM1A] - Loading Control (<a href='/en-us/products/primary-antibodies/alpha-tubulin-antibody-dm1a-loading-control-ab7291'>ab7291</a>) at 1 µg/mL

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma cell line) whole cell lysate at 10 µg

Lane 2:

NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate at 10 µg

Lane 3:

PC12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 dilution

Predicted band size: 36 kDa,50 kDa

Observed band size: 50 kDa

true

Exposure time: 150s

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

CD45RO contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

All lanes:

Western blot - Anti-CD45RO antibody [UCHL1] (<a href='/en-us/products/primary-antibodies/cd45ro-antibody-uchl1-ab23'>ab23</a>) at 5 µg/mL

Lane 1:

Human tonsil normal tissue lysate - total protein (<a href='/en-us/products/unavailable/human-tonsil-normal-tissue-lysate-total-protein-ab29615'>ab29615</a>) at 20 µg

Lane 2:

Human thymus tissue lysate - total protein (<a href='/en-us/products/unavailable/human-thymus-tissue-lysate-total-protein-ab30146'>ab30146</a>) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 147 kDa

Observed band size: 180 kDa

true

Exposure time: 8min

• WB

Lab

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

All lanes:

Western blot - Anti-Tubulin antibody [DM1A +DM1B] - Loading Control (<a href='/en-us/products/primary-antibodies/tubulin-antibody-dm1a-dm1b-loading-control-ab44928'>ab44928</a>) at 5 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Lane 2:

NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 20 µg

Lane 3:

PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 50 kDa

Observed band size: 53 kDa

true

Exposure time: 2min

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

All lanes:

Western blot - Anti-Factor XIIIa antibody [AC-1A1] (<a href='/en-us/products/primary-antibodies/factor-xiiia-antibody-ac-1a1-ab1834'>ab1834</a>) at 1/250 dilution

All lanes:

Human placenta tissue lysate - total protein at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 83 kDa

Observed band size: 37 kDa,74 kDa,83 kDa

true

Exposure time: 2min

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

Podoplanin protein is extensively glycosylated. This might explain the observation of a band at a higher molecular weight than predicted.

All lanes:

Western blot - Anti-Podoplanin antibody [18H5] - BSA and Azide free (<a href='/en-us/products/primary-antibodies/podoplanin-antibody-18h5-bsa-and-azide-free-ab10288'>ab10288</a>) at 5 µg/mL

Lane 1:

Human placenta tissue lysate - total protein (<a href='/en-us/products/unavailable/human-placenta-tissue-lysate-total-protein-ab29745'>ab29745</a>) at 20 µg

Lane 2:

Human skeletal muscle tissue lysate - total protein (<a href='/en-us/products/unavailable/human-skeletal-muscle-tissue-lysate-total-protein-ab29330'>ab29330</a>) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 16 kDa

Observed band size: 35 kDa

true

Exposure time: 20min

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

All lanes:

Western blot - Anti-hnRNP L antibody [4D11] (<a href='/en-us/products/primary-antibodies/hnrnp-l-antibody-4d11-ab6106'>ab6106</a>) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

Lane 4:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 64 kDa

Observed band size: 60 kDa,64 kDa,68 kDa

true

Exposure time: 1min

• WB

Lab

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

This blot was produced using a 4-12% Bis-Tris gel under the MOPS buffer system under denaturing, reducing conditions. The gel was run at 200V for 60 minutes before being transferred onto a nitrocellulose membrane at 30V for 70 minutes. After transfer, the membrane was blocked for an hour in 3% milk before being incubated overnight at 4°C with mouse monoclonal [H4B4] to LAMP2 (ab25631; diluted 1 : 5000). Antibody binding was detected using peroxidase labelled goat anti-mouse IgG (ab97040; diluted 1 : 50000) for an hour at room temperature and visualised using ECL development solution.

All lanes:

Western blot - Anti-LAMP2 antibody [H4B4] - Lysosome Marker (<a href='/en-us/products/primary-antibodies/lamp2-antibody-h4b4-lysosome-marker-ab25631'>ab25631</a>) at 1/500 dilution

Lane 1:

HeLa cell lysate at 20 µg

Lane 2:

Jurkat cell lysate at 20 µg

Lane 3:

Human liver lysate at 20 µg

Lane 4:

Human liver membrane fraction lysate at 20 µg

Lane 5:

Human skeletal muscle lysate at 20 µg

Lane 6:

Human brain lysate at 20 µg

Lane 7:

Human kidney lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/50000 dilution

Predicted band size: 45 kDa

Observed band size: 100 kDa,110 kDa,120 kDa

false

Exposure time: 20min

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-NR2F2 antibody [H7147] (<a href='/en-us/products/primary-antibodies/nr2f2-antibody-h7147-ab41859'>ab41859</a>) at 1 µg/mL

Lane 1:

Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 46 kDa

Observed band size: 49 kDa,72 kDa

true

Exposure time: 4min

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

ab89119 recognizes the recombinant full length soluble Human Amphiregulin protein (ab104355) which has an expected molecular weight of ~12 kDa.

All lanes:

Western blot - Anti-Amphiregulin antibody [MM0089-8K16] (<a href='/en-us/products/primary-antibodies/amphiregulin-antibody-mm0089-8k16-ab89119'>ab89119</a>) at 1/500 dilution

All lanes:

Western blot - Recombinant human Amphiregulin protein (<a href='/en-us/products/proteins-peptides/recombinant-human-amphiregulin-protein-ab104355'>ab104355</a>) at 0.1 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 28 kDa

true

Exposure time: 8min

• WB

Project

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

All lanes:

Western blot - Anti-Vimentin antibody [V9] - Cytoskeleton Marker (<a href='/en-us/products/primary-antibodies/vimentin-antibody-v9-cytoskeleton-marker-ab8069'>ab8069</a>) at 1 µg/mL

Lane 1:

HeLa (Human epithelial cell line from cervix adenocarcinoma) Whole Cell Lysate at 10 µg

Lane 2:

MOLT4 (Human lymphoblastic leukemia cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 53 kDa

Observed band size: 57 kDa

true

Exposure time: 20min

• WB

Lab

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

We believe that the band observed at approximately 33-kDa is a C-terminal cleavage fragment of URP2/Kindlin-3, which has been described in the literature (PMID 23012377). Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

All lanes:

Western blot - Anti-URP2/Kindlin-3 antibody [3D6] (<a href='/en-us/products/primary-antibodies/urp2-kindlin-3-antibody-3d6-ab173416'>ab173416</a>) at 5 µg/mL

Lane 1:

Human spleen tissue lysate - total protein (<a href='/en-us/products/unavailable/human-spleen-tissue-lysate-total-protein-ab29699'>ab29699</a>) at 20 µg

Lane 2:

Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate at 20 µg

Lane 3:

K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate at 20 µg

Lane 4:

THP1 (Human acute monocytic leukemia cell line) Whole Cell Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

Predicted band size: 75 kDa

Observed band size: 33 kDa,76 kDa

true

Exposure time: 4min

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-HSD3B1 antibody [3C11-D4] (<a href='/en-us/products/primary-antibodies/hsd3b1-antibody-3c11-d4-ab55268'>ab55268</a>) at 1 µg/mL

All lanes:

Human placenta tissue lysate - total protein (<a href='/en-us/products/unavailable/human-placenta-tissue-lysate-total-protein-ab29745'>ab29745</a>) at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa,73 kDa

true

Exposure time: 4min

• WB

Lab

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

All lanes:

Western blot - Anti-Cyclin A2 antibody [E23.1] (<a href='/en-us/products/primary-antibodies/cyclin-a2-antibody-e231-ab38'>ab38</a>) at 5 µg/mL

Lane 1:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 20 µg

Lane 2:

U2OS (Human osteosarcoma cell line) Whole Cell Lysate at 20 µg

Lane 3:

PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 20 µg

Lane 4:

NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

Predicted band size: 49 kDa

Observed band size: 50 kDa

true

Exposure time: 20min

• WB

Project

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

Abcam recommends using milk (5%) as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-arginine and sodium azide (ab13575).

All lanes:

Western blot - Anti-Cytochrome C antibody [7H8.2C12] (<a href='/en-us/products/primary-antibodies/cytochrome-c-antibody-7h82c12-ab13575'>ab13575</a>) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

Lane 3:

Human heart tissue lysate - total protein (<a href='/en-us/products/unavailable/human-heart-tissue-lysate-total-protein-ab29431'>ab29431</a>) at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 11 kDa

Observed band size: 14 kDa

true

Exposure time: 3min

• WB

Project

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

All lanes:

Western blot - Anti-Nucleophosmin antibody [3A9F1] (<a href='/en-us/products/primary-antibodies/nucleophosmin-antibody-3a9f1-ab86712'>ab86712</a>) at 10 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

U2OS (Human osteosarcoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 32 kDa

Observed band size: 100 kDa,150 kDa,37 kDa,55 kDa

true

Exposure time: 1min

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

This image was generated using the ascites version of the product.

All lanes:

Western blot - Anti-HNF-4-alpha antibody [K9218] (<a href='/en-us/products/primary-antibodies/hnf-4-alpha-antibody-k9218-ab41898'>ab41898</a>) at 1 µg/mL

All lanes:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 53 kDa

Observed band size: 108 kDa,37 kDa,53 kDa

true

Exposure time: 4min

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

The band observed at 52 kDa could potentially be a cleaved form of Angiopoietin 2/ANG2 due to the presence of an 18 amino acid signal peptide.

All lanes:

Western blot - Anti-Angiopoietin 2/ANG2 antibody [MM0020-1F29] (<a href='/en-us/products/primary-antibodies/angiopoietin-2-ang2-antibody-mm0020-1f29-ab56301'>ab56301</a>) at 1 µg/mL

All lanes:

Human liver tissue lysate - total protein (<a href='/en-us/products/unavailable/human-liver-tissue-lysate-total-protein-ab29889'>ab29889</a>) at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 56 kDa

Observed band size: 47 kDa,52 kDa,65 kDa

true

Exposure time: 4min

• WB

Project

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

The predicted molecular weight of Acid Sphingomyelinase protein is 70 kDa. However, the protein sequence contains a 46-residue signal sequence at the amino-terminal, which could explain the band observed.

All lanes:

Western blot - Anti-Acid sphingomyelinase antibody [mAbcam74281] (<a href='/en-us/products/primary-antibodies/acid-sphingomyelinase-antibody-mabcam74281-ab74281'>ab74281</a>) at 5 µg/mL

Lane 1:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

A431 (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 3:

MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 70 kDa

Observed band size: 65 kDa

true

Exposure time: 1min

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

NFAT1 contains an exstensive number of potential phosphorylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.

All lanes:

Western blot - Anti-NFAT1 antibody [25A10.D6.D2] (<a href='/en-us/products/primary-antibodies/nfat1-antibody-25a10d6d2-ab2722'>ab2722</a>) at 1 µg/mL

All lanes:

Human spleen tissue lysate - total protein (<a href='/en-us/products/unavailable/human-spleen-tissue-lysate-total-protein-ab29699'>ab29699</a>) at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 100 kDa

Observed band size: 150 kDa,62 kDa

true

Exposure time: 4min

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

The 120 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to Ctip1 / BCL-11A.

All lanes:

Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

true

Exposure time: 20min

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

All lanes:

Western blot - Anti-PAX7 antibody [mAbcam61067] (<a href='/en-us/products/primary-antibodies/pax7-antibody-mabcam61067-ab61067'>ab61067</a>) at 2 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Lane 2:

NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 56 kDa

Observed band size: 37 kDa,40 kDa,55 kDa,65 kDa,70 kDa

true

Exposure time: 2min

• WB

Project

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

All lanes:

Western blot - Anti-PP2A alpha + beta (methyl L309) antibody [2A10] (<a href='/en-us/products/primary-antibodies/pp2a-alpha-beta-methyl-l309-antibody-2a10-ab66597'>ab66597</a>) at 10 µg/mL

Lane 1:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 20 µg

Lane 2:

PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

Predicted band size: 36 kDa

Observed band size: 36 kDa

true

Exposure time: 20min

• WB

Project

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

All lanes:

Western blot - Anti-PROX1 antibody [5G10] (<a href='/en-us/products/primary-antibodies/prox1-antibody-5g10-ab33219'>ab33219</a>) at 10 µg/mL

Lane 1:

Western blot - Hep G2 nuclear extract lysate (<a href='/en-us/products/tissue-lysates/hep-g2-nuclear-extract-lysate-ab14660'>ab14660</a>) at 20 µg

Lane 2:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate (negative control) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 83 kDa

Observed band size: 83 kDa

true

Exposure time: 20min

• WB

Project

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

We recommend using 3% milk as the blocking agent for Western blot.

All lanes:

Western blot - Anti-beta Catenin antibody [12F7] (<a href='/en-us/products/primary-antibodies/beta-catenin-antibody-12f7-ab22656'>ab22656</a>) at 2 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Lane 2:

DU 145 (Human prostate carcinoma cell line) Whole Cell Lysate at 20 µg

Lane 3:

A549 (Human lung adenocarcinoma epithelial cell line) Whole Cell Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 85 kDa

Observed band size: 85 kDa

true

Exposure time: 12min

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

All lanes:

Western blot - Anti-Calbindin antibody [AF2E5] (<a href='/en-us/products/primary-antibodies/calbindin-antibody-af2e5-ab75524'>ab75524</a>) at 1/500 dilution

Lane 1:

Human kidney tissue lysate - total protein (<a href='/en-us/products/unavailable/human-kidney-tissue-lysate-total-protein-ab30203'>ab30203</a>) at 10 µg

Lane 2:

Kidney (Rat) Tissue Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 30 kDa

Observed band size: 30 kDa

true

Exposure time: 90s

• WB

Project

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

All lanes:

Western blot - Anti-alpha Tubulin (acetyl K40) antibody [6-11B-1] (<a href='/en-us/products/primary-antibodies/alpha-tubulin-acetyl-k40-antibody-6-11b-1-ab24610'>ab24610</a>) at 5 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Lane 2:

NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 20 µg

Lane 3:

PC12 (Rat adrenal pheochromocytoma cell line) Whole Cell Lysate at 20 µg

Lane 4:

Brain (Mouse) Tissue Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 50 kDa

Observed band size: 140 kDa,25 kDa,35 kDa,55 kDa

false

Exposure time: 150s

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

All lanes:

Western blot - Anti-PAX5 antibody [mAbcam52588] (<a href='/en-us/products/primary-antibodies/pax5-antibody-mabcam52588-ab52588'>ab52588</a>) at 1 µg/mL

Lane 1:

Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate (blocked with 1% milk) at 20 µg

Lane 2:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate (blocked with 1% Milk) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

Predicted band size: 42 kDa

Observed band size: 45 kDa,50 kDa

true

Exposure time: 20min

• WB

Lab

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

All lanes:

Western blot - Anti-Y14 antibody [4C4] (<a href='/en-us/products/primary-antibodies/y14-antibody-4c4-ab5828'>ab5828</a>) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

Lane 3:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 4:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 19 kDa

Observed band size: 21 kDa,53 kDa

true

Exposure time: 8min

• WB

Project

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

Blocked using 3% milk.

All lanes:

Western blot - Anti-HSD3B2 antibody [373CT9.1.3] (<a href='/en-us/products/primary-antibodies/hsd3b2-antibody-373ct913-ab75710'>ab75710</a>) at 1 µg/mL

All lanes:

Human adrenal normal tissue lysate - total protein (<a href='/en-us/products/unavailable/human-adrenal-normal-tissue-lysate-total-protein-ab29249'>ab29249</a>) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 42 kDa

Observed band size: 110 kDa,35 kDa,40 kDa

true

Exposure time: 20min

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

The band observed at 26 kDa could potentially be a cleaved form of NDUFS3 due to the presence of a 36 amino acid transit peptide.

All lanes:

Western blot - Anti-NDUFS3 antibody [17D95] (<a href='/en-us/products/primary-antibodies/ndufs3-antibody-17d95-ab14711'>ab14711</a>) at 1 µg/mL

Lane 1:

Human brain tissue lysate - total protein (<a href='/en-us/products/unavailable/human-brain-tissue-lysate-total-protein-ab29466'>ab29466</a>) at 10 µg

Lane 2:

Human heart tissue lysate - total protein (<a href='/en-us/products/unavailable/human-heart-tissue-lysate-total-protein-ab29431'>ab29431</a>) at 10 µg

Lane 3:

Heart (Mouse) Tissue Lysate at 10 µg

Lane 4:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 30 kDa

Observed band size: 26 kDa,57 kDa

true

Exposure time: 1min

• WB

Project

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

Abcam recommends using milk as the blocking agent when running western blots of ab80039. This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% milk before being incubated with ab80039 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-AMPK alpha 1 + AMPK alpha 2 antibody [34.2] (<a href='/en-us/products/primary-antibodies/ampk-alpha-1-ampk-alpha-2-antibody-342-ab80039'>ab80039</a>) at 1 µg/mL

Lane 1:

Heart (Rat) Tissue Lysate at 10 µg

Lane 2:

Human brain tissue lysate - total protein (<a href='/en-us/products/unavailable/human-brain-tissue-lysate-total-protein-ab29466'>ab29466</a>) at 10 µg

Lane 3:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 4:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

Predicted band size: 64 kDa

Observed band size: 62 kDa,75 kDa,80 kDa

true

Exposure time: 20min

• WB

Project

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

This data was developed using the same antibody clone in a different buffer formulation containing PBS, L-Arginine and sodium azide (ab26).

All lanes:

Western blot - Anti-p53 antibody [PAb 240] (<a href='/en-us/products/primary-antibodies/p53-antibody-pab-240-ab26'>ab26</a>) at 5 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Lane 2:

Hela Whole Cell Lysate - Bleomycin Treated (40U/ml) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 43 kDa

true

Exposure time: 4min

• WB

Ap

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 3% Milk before being incubated with ab11315 overnight at 4°C. Antibody binding was detected using an anti-mouse antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-beta IV Tubulin antibody [ONS.1A6] (<a href='/en-us/products/primary-antibodies/beta-iv-tubulin-antibody-ons1a6-ab11315'>ab11315</a>) at 5 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Lane 2:

HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 20 µg

Lane 3:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 20 µg

Lane 4:

K562 (Human erythromyeloblastoid leukemia cell line) Whole Cell Lysate at 20 µg

Lane 5:

NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate at 20 µg

Lane 6:

U2OS (Human osteosarcoma cell line) Whole Cell Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/10000 dilution

Predicted band size: 50 kDa

Observed band size: 52 kDa

true

Exposure time: 4min

• WB

Project

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

Abcam recommends using milk as the blocking agent. Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

All lanes:

Western blot - Anti-Cathepsin L + V antibody [33/2] (<a href='/en-us/products/primary-antibodies/cathepsin-l-v-antibody-33-2-ab6314'>ab6314</a>) at 1/1000 dilution

Lane 1:

Lung (Human) Tissue Lysate at 20 µg

Lane 2:

Lung (Mouse) Tissue Lysate at 20 µg

Lane 3:

Human kidney tissue lysate - total protein (<a href='/en-us/products/unavailable/human-kidney-tissue-lysate-total-protein-ab30203'>ab30203</a>) at 20 µg

Lane 4:

Kidney (Mouse) Tissue Lysate at 20 µg

Lane 5:

A549 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 38 kDa

Observed band size: 25 kDa,55 kDa

true

Exposure time: 20min

• WB

Project

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

All lanes:

Western blot - Anti-Coilin antibody [IH10] (<a href='/en-us/products/primary-antibodies/coilin-antibody-ih10-ab87913'>ab87913</a>) at 5 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 2:

HeLa (Human epithelial carcinoma cell line) Nuclear Lysate at 10 µg

Lane 3:

Jurkat (Human T cell lymphoblast-like cell line) Whole Cell Lysate at 10 µg

Lane 4:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 5:

HEK293 (Human embryonic kidney cell line) Whole Cell Lysate at 10 µg

Lane 6:

MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate at 10 µg

Lane 7:

Caco 2 (Human colonic carcinoma cell line) Whole Cell Lysate at 10 µg

Lane 8:

SHSY-5Y (Human neuroblastoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 62 kDa

Observed band size: 28 kDa,75 kDa

true

Exposure time: 20min

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

The 120 kDa band observed is comparable to the molecular weight seen with other commercially available antibodies to Ctip1 / BCL-11A.

All lanes:

Western blot - Anti-Ctip1/BCL-11A antibody [14B5] (<a href='/en-us/products/primary-antibodies/ctip1-bcl-11a-antibody-14b5-ab19487'>ab19487</a>) at 1 µg/mL

All lanes:

Raji (Human Burkitt's lymphoma cell line) Whole Cell Lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 91 kDa

Observed band size: 120 kDa,37 kDa,42 kDa,62 kDa

true

Exposure time: 20min

• WB

Project

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

Blocked using 3% milk.

All lanes:

Western blot - Anti-Alpha Skeletal Muscle Actin antibody [337CT 30.10.1] - Loading Control (<a href='/en-us/products/primary-antibodies/alpha-skeletal-muscle-actin-antibody-337ct-30101-loading-control-ab88226'>ab88226</a>) at 5 µg/mL

Lane 1:

Heart (Mouse) Tissue Lysate at 20 µg

Lane 2:

Heart (Rat) Tissue Lysate at 20 µg

Lane 3:

Human skeletal muscle tissue lysate - total protein (<a href='/en-us/products/unavailable/human-skeletal-muscle-tissue-lysate-total-protein-ab29330'>ab29330</a>) at 20 µg

Lane 4:

Skeletal Muscle (Mouse) Tissue Lysate at 20 µg

Lane 5:

Skeletal Muscle (Rat) Tissue Lysate at 20 µg

Lane 6:

TE 671 (Human Rhabdomyosarcoma) Whole Cell Lysate at 20 µg

Lane 7:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 42 kDa

Observed band size: 42 kDa

true

Exposure time: 30s

• WB

Unknown

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (AB97040)

All lanes:

Western blot - Anti-Elastin antibody [BA-4] (<a href='/en-us/products/primary-antibodies/elastin-antibody-ba-4-ab9519'>ab9519</a>) at 0.51 µg/mL

All lanes:

Human skin tissue lysate - total protein (<a href='/en-us/products/unavailable/human-skin-tissue-lysate-total-protein-ab30166'>ab30166</a>) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution

Predicted band size: 68 kDa

Observed band size: 55 kDa,70 kDa

true

Exposure time: 20min