Membrane Fraction WB Cocktail (ab140365)
Key features and details
- Sample type: Cell culture extracts, Cell Lysate, Tissue Extracts
Overview
-
Product name
Membrane Fraction WB Cocktail -
Sample type
Cell culture extracts, Tissue Extracts, Cell Lysate -
Species reactivity
Reacts with: Mouse, Rat, Human -
Product overview
ab140365 contains 5 mAbs each targeting proteins located in different compartments of the cell. The presence of plasma membrane is shown by Anti-Sodium Potassium ATPase; endoplasmic reticulum is detected by GRP78; mitochondrial by ATP5A; cytosol by Anti-GAPDH; and nucleus by Anti-Histone H3 (di methyl K9). This cocktail is suitable for detecting the purity of cellular fractions using kit ab139409.
-
Tested applications
Suitable for: WBmore details
Properties
-
Storage instructions
Store at -20°C. Please refer to protocols. -
Components 200 µl 2500X HRP Conjugated Secondary Antibody Cocktail 1 x 60µl 250X Membrane Fraction Western Blot Cocktail 1 x 200µl -
Research areas
Associated products
-
Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab140365 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
| Application | Abreviews | Notes |
|---|---|---|
| WB |
Use at an assay dependent concentration.
Primary antibody cocktail = 1/250 dilution Secondary antibody cocktail = 1/2500 dilution |
| Notes |
|---|
|
WB
Use at an assay dependent concentration. Primary antibody cocktail = 1/250 dilution Secondary antibody cocktail = 1/2500 dilution |
Images
-
Membrane Fraction Western Blot Cocktail - Component SeperationPerformed under reducing conditions. All blocking and antibody incubation steps were done in 5% milk in TBST. Developed using the ECL technique. Exposure time: 5 mins.
Sample 1: Marker
Samples 2-5: Hela Cell Lysate – 20 µg
Primary:
Lane 1: none
Lane 2: Anti- Sodium Potassium ATPase antibody – Plasma Membrane Marker
Lane 3: Anti-GRP78 antibody – Endoplasmic Reticulum Marker
Lane 4: Anti- ATP5a antibody – Mitochondrial Membrane Marker
Lane 5: Anti- GAPDH antibody – Cytosolic Marker
Lane 6: Anti- Histone H3 (di methyl K9) antibody – Nuclear Marker
Lane 7: Assembled Membrane Antibody Cocktail
Secondary:HRP conjugated secondary antibody cocktail
Predicted/observed band sizes:
Sodium Potassium ATPase = 112 kDa/100 kDa
GRP78 band size: 78 kDa / 75 kDa
ATP5A band size: 60 kDa / 60 kDa
GAPDH band size: 37 kDa / 38 kDa
Histone H3 (di methyl K9) band size: 17 kDa / 17 kDa -
Membrane Fraction Western Blot Cocktail - SpecificityDeveloped by IR scanning. HeLa cell lysate prepared using Kit ab139409. Exposure time: 5 mins . Samples run on a 4-20% gradient acrylamide gel. All blocking and antibody incubation steps were done in 5% milk, 20mM Tris-HCl, 0.1% TWEEN-20
Lane 1: Marker
Lane 2 : HeLa Whole Cell Lysate
Lane 3 : HeLa Cytosolic Fraction Lysate
Lane 4 : HeLa Membrane Fraction Lysate
Lane 5 : HeLa Nuclear Fraction Lysate
Primary antibody: Membrane Fraction Western Blot cocktail
Secondary: IR800 GAM (1/10000) and IR690 GAR (1/10000)
Predicted/Observed band sizes:
Sodium Potassium ATPase: 112 kDa/100 kDa
GRP78: 78 kDa/75 kDa
ATP5A: 60 kDa/60 kDa
GAPDH: 37 kDa/37kDa
Histone H3 (di methyl K9): 17 kDa/17kDa
Notes: Percentage of antibody signal (Y axis) represents the signal present in individual fractions as a proportion of the sum of all three fractions (cytoplasmic, membrane and nuclear) -
Membrane Fraction Western Blot Cocktail - SpecificityDeveloped using ECL technique under reducing conditions. HeLa lysate prepared using the Membrane Fractionation Kit (ab139409). Exposure time: 5 mins. on a 4-20% gradient acrylamide gel, blocking and antibody incubation steps done in 5% milk, 20mM Tris-HCl, 0.1% TWEEN-20
Lane 1: Marker
Lane 2 : HeLa Whole Cell Lysate
Lane 3 : HeLa Cytosolic Fraction Lysate
Lane 4 : HeLa Membrane Fraction Lysate
Lane 5 : HeLa Nuclear Fraction Lysate
Primary antibody: Membrane Fraction Western Blot cocktail
Secondary: HRP conjugated secondary antibody cocktail
Predicted/Observed band sizes:
Sodium Potassium ATPase: 112 kDa/100 kDa
GRP78: 78 kDa/75 kDa
ATP5A: 60 kDa/60 kDa
GAPDH: 37 kDa/37kDa
Histone H3 (di methyl K9): 17 kDa/17kDa
Notes: Percentage of antibody signal represents signal present in individual fractions as a proportion of the sum of all three fractions (cytoplasmic, membrane and nuclear) -
Membrane Fraction Western Blot Cocktail - Cross ReactivitySamples run on a 4-20% gradient acrylamide gel. Performed under reducing conditions. All blocking and antibody incubation steps were done in 5% milk, 20mM Tris-HCl, 0.1% TWEEN-20. Developed using the ECL technique. Exposure time: 5 mins.
Lane 1: Marker
Lane 2: Human heart homogenate Lysate – 20 µg
Lane 3: Hela Cell Lysate – 20 µg
Lane 4: Mouse heart homogenate Lysate - 20 µg
Lane 5: NIH3T3 Cell Lysate – 20 µg
Lane 6: Rat heart homogenate Lysate – 20 µg
Lane 7: H9C2 Cell Lysate – 20 µg
Primary antibody: Membrane Fraction Western blot cocktail at 1/250 dilution
Secondary antibody: HRP conjugated secondary antibody cocktail at 1/2500 dilution
Predicted/observed band sizes:
Sodium Potassium ATPase = 112 kDa/100 kDa
GRP78 band size: 78 kDa / 75 kDa
ATP5A band size: 60 kDa / 60 kDa
GAPDH band size: 37 kDa / 38 kDa
Histone H3 (di methyl K9) band size: 17 kDa / 17 kDa
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
References (2)
ab140365 has been referenced in 2 publications.
- Foryst-Ludwig A et al. Adipose Tissue Lipolysis Promotes Exercise-induced Cardiac Hypertrophy Involving the Lipokine C16:1n7-Palmitoleate. J Biol Chem 290:23603-15 (2015). WB ; Mouse . PubMed: 26260790
- Bohuslavova R et al. Partial deficiency of HIF-1a stimulates pathological cardiac changes in streptozotocin-induced diabetic mice. BMC Endocr Disord 14:11 (2014). WB ; Mouse . PubMed: 24502509
