Rabbit Loading Control Antibody Panel

• WB

Lab

Western blot - Rabbit Loading Control Antibody Panel (AB199712)

All lanes : Anti-Histone H3 antibody [EPR16987] - Nuclear Loading Control and ChIP Grade (ab176842) at 1 μg

Lane 1 : HeLa (Human epithelial carcinoma 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 : Drosophila embryo nuclear extract (from melanogaster embryos 0-12hr) at 10 μg
Lane 4 : S.cerevisiae (Y190) Whole Cell Lysate at 20 μg
Lane 5 : S.pombe Whole Cell Lysate at 20 μg

Secondary
Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/50000 dilution
Developed using the ECL technique

Performed under reducing conditions.

Predicted band size : 15 kDa
Observed band size : 17 kDa (why is the actual band size different from the predicted?)


Exposure time : 5 seconds

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 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 ab176842 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406.

All lanes:

Western blot - Anti-Histone H3 antibody [EPR16987] - Nuclear Marker and ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h3-antibody-epr16987-nuclear-marker-and-chip-grade-ab176842'>ab176842</a>) at 1 µg

Lane 1:

HeLa (Human epithelial carcinoma 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:

Drosophila embryo nuclear extract (from melanogaster embryos 0-12hr) at 10 µg

Lane 4:

S.cerevisiae (Y190) Whole Cell Lysate at 20 µg

Lane 5:

S.pombe Whole Cell Lysate at 20 µg

Secondary

All lanes:

Peroxidase AffiniPure Goat Anti-Rabbit IgG (H+L) at 1/50000 dilution

Predicted band size: 15 kDa

Observed band size: 17 kDa

true

Exposure time: 5s

• WB

Unknown

Western blot - Rabbit Loading Control Antibody Panel (AB199712)

All lanes : Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (ab76020) at 1/100000 dilution

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysates
Lane 3 : 293 (Human embryonic kidney epithelial cell) whole cell lysates
Lane 4 : A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates

Lysates/proteins at 20 μg per lane.

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution

Predicted band size : 113 kDa
Observed band size : 100 kDa (why is the actual band size different from the predicted?)


Exposure time : 2 minutes

Blocking and diluting buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Sodium Potassium ATPase antibody [EP1845Y] - Plasma Membrane Loading Control (<a href='/en-us/products/primary-antibodies/sodium-potassium-atpase-antibody-ep1845y-plasma-membrane-loading-control-ab76020'>ab76020</a>) at 1/100000 dilution

Lane 1:

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

Lane 2:

MCF-7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 20 µg

Lane 3:

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

Lane 4:

A431 (Human epidermoid carcinoma epithelial cell) whole cell lysates at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/20000 dilution

Predicted band size: 113 kDa

Observed band size: 100 kDa

false

Exposure time: 2min

• WB

Supplier Data

Western blot - Rabbit Loading Control Antibody Panel (AB199712)

All lanes : Anti-alpha Tubulin antibody [EPR13478(B)] - Loading Control (ab176560) at 1/1000 dilution (Unpurified)

Lane 1 : HeLa cell lysate
Lane 2 : Jurkat cell lysate
Lane 3 : A431 cell lysate
Lane 4 : K562 cell lysate

Lysates/proteins at 10 μg per lane.

Developed using the ECL technique

Predicted band size : 50 kDa

Secondary antibodies - anti-rabbit HRP (ab6721)

All lanes:

Western blot - Anti-alpha Tubulin antibody [EPR13478(B)] - Loading Control (<a href='/en-us/products/primary-antibodies/alpha-tubulin-antibody-epr13478b-loading-control-ab176560'>ab176560</a>) at 1/1000 dilution

Lane 1:

HeLa cell lysate at 10 µg

Lane 2:

Jurkat cell lysate at 10 µg

Lane 3:

A431 cell lysate at 10 µg

Lane 4:

K562 cell lysate at 10 µg

Predicted band size: 50 kDa

true

• WB

Supplier Data

Western blot - Rabbit Loading Control Antibody Panel (AB199712)

All lanes : Anti-GAPDH antibody [EPR16891] (ab181602) at 1/50000 dilution

Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
Lane 2 : Xenopus(X. tropicalis) muscle lysates
Lane 3 : UMNSAH/DF-1 (Transformed chicken embyronic fibroblast cells) whole cell lysates
Lane 4 : Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates

Lysates/proteins at 20 μg per lane.

Secondary
Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size : 36 kDa
Observed band size : 36 kDa

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-GAPDH antibody [EPR16891] - Loading Control (<a href='/en-us/products/primary-antibodies/gapdh-antibody-epr16891-loading-control-ab181602'>ab181602</a>) at 1/50000 dilution

Lane 1:

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

Lane 2:

Xenopus tropicalis muscle lysates at 20 µg

Lane 3:

UMNSAH/DF-1 (Transformed chicken embyronic fibroblast cells) whole cell lysates at 20 µg

Lane 4:

Jurkat (Human T cell leukemia cells from peripheral blood) whole cell lysates at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

Predicted band size: 36 kDa

Observed band size: 36 kDa

false

• WB

Supplier Data

Western blot - Rabbit Loading Control Antibody Panel (AB199712)

All lanes : Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control (ab202554) at 1/10000 dilution

Lane 1 : Mouse heart lysate
Lane 2 : Rat heart lysate

Lysates/proteins at 10 μg per lane.

Secondary
Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size : 20 kDa
Observed band size : 17 kDa (why is the actual band size different from the predicted?)


Exposure time : 3 minutes

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-COX IV antibody [EPR9442(ABC)] - Mitochondrial Loading Control (<a href='/en-us/products/primary-antibodies/cox-iv-antibody-epr9442abc-mitochondrial-loading-control-ab202554'>ab202554</a>) at 1/10000 dilution

Lane 1:

Mouse heart lysate at 10 µg

Lane 2:

Rat heart lysate at 10 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

Predicted band size: 19 kDa

Observed band size: 17 kDa

false

Exposure time: 3min