Western blot abreview for Anti-Collagen I antibody

Good
Abreviews
Application
Western blot
Sample
Salmo salar Tissue lysate - whole (Skin)
Gel Running Conditions
Reduced Denaturing (6%)
Loading amount
10 µg
Treatment
500 mM acetic acid
Specification
Skin
Blocking step
No blocking step used for 1 hour(s) and 0 minute(s) · Temperature: 22°C

Other product details

Incubation time
18 hour(s) and 0 minute(s) · Temperature: 4°C · Diluent: PBS-T
Dilution
1/10000

Secondary antibody

Dilution
1/10000
Name
Non-Abcam antibody was used: goat anti-rabbit IgG antibody HRP (Vector Labs)
Host species: Rabbit
Clonality: Polyclonal
Conjugation: Horse Radish Peroxidase

Detection

Detection method
ECL
Negative control
Second blot treated with PBS-T instead of primary antibody (ab34710)
Exposure
1 minute(s) and 0 second(s)
Bands
Specific: 127, 136 kDa Non-specific: 70, 90, 100, >180 kDa
Positive control
No positive control

Additional data

Additional Notes
ad TREATMENT) Cooked skin (gelatin) dissolved in 500 mM acetic acid. Dialyzed against water after separation from unsoluable components. (Gelatin was treated with acetic acid to solubilize it) ad BLOCKING AGENT) 1 hour treatment with PBS-T (no BSA or else) ad BLOCKING TEMPERATURE) Room temperature: 22°C ad BANDS) bands at 136 kDa and 127 kDa refer to the collagen type-I alpha-1 chain and alpha-2 chain (mass spectrometry analysis). Bands above 180 kDa may represent oligomeric collagen type-I. ad POSITIVE CONTROL) MCF-7 lysate was planned as positive control, but the lysate appeared as smear on the gel. Thus, the positive control was left. ad IMAGE) all samples: Cooked skin (gelatin) from Salmo salar 1) gelatin, estimated amount: ~10 µg (prim ab: ab34710) 2) extract, 10 µg, treated with 500 mM acetic acid (prim ab: ab34710) 3) gelatin, estimated amount: ~10 µg (negative control) 4) extract, 10 µg, treated with 500 mM acetic acid 10 µg (negative control) lanes 1 and 3: loaded with ~10 µg protein (gelatin directly loaded onto gel)

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Submitted Jun 20 2018

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