Endoplasmic Reticulum Fraction Western Blot Cocktail
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(2 Publications)
- WB
Supplier Data
Western blot - Endoplasmic Reticulum Fraction Western Blot Cocktail (AB139415)
Developed using the ECL technique, performed under reducing conditions and exposed for 3 minutes. All blocking and antibody incubation steps performed in 5% milk, 20 mM Tris-HCl, 0.1% TWEEN-20.
Samples :
1 : Marker
2 : Human heart homogenate lysate - 20 µg
3 : HeLa cell lysate - 20 µg
4 : Mouse heart homogenate lysate - 20 µg
5 : NIH3T3 cell lysate - 20 µg
6 : Rat heart homogenate lysate - 20 µg
7 : H9C2 cell lysate - 20 µg
Primary antibody : ER Fraction WB cocktail at 1/250.
Secondary antibody : HRP conjugated secondary antibody cocktail at 1/2500.
Predicted/Observed band sizes :
GRP78 : 78 kDa/75 kDa
GAPDH : 37 kDa/37 kDa
Histone H3 (dimethyl K9) : 17 kDa/17 kDa
false
- WB
Supplier Data
Western blot - Endoplasmic Reticulum Fraction Western Blot Cocktail (AB139415)
Developed using the ECL technique; Performed under reducing conditions; Exposure time : 5 mins. All blocking and antibody incubation steps were performed in 5% milk, 20 mM Tris-HCl, 0.1% TWEEN-20
Samples
Lane 1 : Marker
Lanes 2-5 : Hela Whole Cell Lysate – 20 µg
Primary antibody :
Lane 1 : None
Lane 2 : Anti-GRP78 antibody – Endoplasmic Reticulum Marker
Lane 3 : Anti- GAPDH antibody – Cytosolic Marker
Lane 4 : Anti- Histone H3 (di methyl K9) antibody – Nuclear Marker
Lane 5 : Assembled Endoplasmic Reticulum Antibody Cocktail
Secondary : HRP conjugated secondary antibody cocktail at 1/2500
Predicted GRP78 band size : 78 kDa
Observed band size : 78 kDa
Predicted GAPDH band size : 37 kDa
Observed band size : 37 kDa
Predicted Histone H3 (di methyl K9) band size : 17 kDa
Observed band size : 17 kDa
false
- WB
Supplier Data
Western blot - Endoplasmic Reticulum Fraction Western Blot Cocktail (AB139415)
HeLa cell lysates were prepared using the Membrane Fractionation Kit (ab139409), blots were developed using the ECL technique, performed under reducing conditions and exposed for 5 minutes. A 4-15% gradient acrylamide gel was used and blocking and antibody incubation steps were performed in 5% milk, 20 mM Tris-HCl, 0.1% TWEEN-20.
Sample lanes :
Lane 1 – Marker
Lane 2 - HeLa whole cell lysate - 20 µL
Lane 3 - HeLa cytosolic fraction lysate - 20 µL
Lane 4 - HeLa membrane fraction lysate - 20 µL
Lane 5 - HeLa nuclear fraction lysate - 20 µL
Primary antibody : ER Fraction WB cocktail at 1/250 dilution
Secondary antibody : HRP conjugated secondary cocktail at 1/2500.
Predicted/observed band sizes :
GRP78 : 78 kDa/78 kDa
GAPDH : 37 kDa/37 kDa
Histone H3 (di methyl K9) : 17 kDa/17 kDa
false
Reactivity data
Product details
ab139415 contains 3 mAbs each targeting a specific organelle marker. The presence of endoplasmic reticulum is detected by GRP78; cytosol by Anti-GAPDH; and nucleus by Anti-Histone H3 (di methyl K9). This cocktail is suitable for determining the purity of organelle isolates prior to further characterization.
This product is particularly valuable to researchers working in organelle proteomics. Mass spectrometry is frequently used in this field to determine the protein content of targeted organelle isolates. These isolates are obtained using differential centrifugation, density gradient fractionation, biochemical enrichment, or affinity purification. Unfortunately, the various methods of purification available for organelle isolation are imperfect and leave behind contaminants from undesired regions of the cell. These contaminants are inevitable, but being aware of which contaminants are present is crucial for analysis of mass spectrometry results. The high sensitivity and species cross reactivity of the antibodies in this cocktail will quickly and easily reveal impurities caused by imperfect sample preparation.
Properties and storage information
Shipped at conditions
Appropriate short-term storage conditions
Appropriate long-term storage conditions
Supplementary information
This supplementary information is collated from multiple sources and compiled automatically.
Biological function summary
GRP78 BiP participates in maintaining cellular homeostasis by regulating the unfolded protein response and aiding in the assembly of multimeric protein complexes. GRP78 interacts closely with various other proteins to mitigate stress in the endoplasmic reticulum such as protein disulfide isomerase. GAPDH is central to energy production and intersects with various cellular processes including autophagy and nuclear functions. Histone H3 contributes to chromatin structure and gene expression functioning as part of the histone octamer complex.
Pathways
GRP78 BiP functions in the endoplasmic reticulum stress pathway connecting with key proteins like ATF6 during the unfolded protein response. GAPDH plays a role in the glycolytic pathway linking to enzymes such as hexokinase and pyruvate kinase facilitating energy production. Histone H3 is important in the DNA replication and repair pathway interacting with proteins such as histone acetyltransferases that modify chromatin structure and gene expression.
Target data
Publications (2)
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PloS one 15:e0237930 PubMed32841274
2020
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Species
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Bio-protocol 9:e3379 PubMed33654875
2019
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Unspecified application
Species
Unspecified reactive species
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