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MAP1LC3A / MAP1LC3B

Cat Number: AB-83557
Size: 100 ul
Clone: POLY
Concentration: 1mg/ml
Host: Rb
Isotype: IgG
Immunogen: Recombinant fusion protein containing a sequence corresponding to amino acids 1-121 of human LC3A
Reactivity: Hu, Ms, Rt
Applications:

Western Blot: 1: 1000
Immunofluorescence: 1:50-200

Molecular: 14-16kDa
Purification: Purified
Synonyms: MAP1LC3B;ATG8F;LC3B;MAP1A/1BLC3;MAP1LC3B-a;MAP1LC3A;ATG8E;LC3;LC3A;MAP1ALC3;MAP1BLC3;LC3A/LC3B
Background:

MAP1A and MAP1B are microtubule-associated proteins which mediate the physical interactions between microtubules and components of the cytoskeleton. MAP1A and MAP1B each consist of a heavy chain subunit and multiple light chain subunits. The protein encoded by this gene is one of the light chain subunits and can associate with either MAP1A or MAP1B. Two transcript variants encoding different isoforms have been found for this gene. The expression of variant 1 is suppressed in many tumor cell lines, suggesting that may be involved in carcinogenesis.

Form: Liquid
Buffer: PBS with 0.02% sodium azide, 50% glycerol pH 7.3
Storage: At -20°C for one year. After reconstitution, at 4°C for one month. It can also be aliquotted and stored frozen at -20°C for a longer time. Avoid repeated freezing and thawing.
AB-83557-1.jpg

Western blot analysis of extracts of various cell lines, using LC3A/LC3B antibody at 1:500 dilution.
Secondary antibody: HRP Goat Anti-Rabbit IgG (H+L) at 1:10000 dilution.
Lysates/proteins: 25ug per lane.
Blocking buffer: 3% nonfat dry milk in TBST.
Detection: ECL West Pico
Exposure time: 20s.

AB-83557-2.jpg

Immunofluorescence analysis of HeLa cells using LC3A/LC3B antibody

AB-83557-3.jpg

Product:
MAP1LC3A/MAP1LC3B
Journal:
Acta Pharmacologica Sinica
Application:
WB, IF
IF:4.01
Species:Mus musculus
PMID:31316178
Title: LW-213, a newly synthesized flavonoid, induces G2/M phase arrest and apoptosis in chronic myeloid leukemia.

Western Blot Protocol All steps are carried out at room temperature unless otherwise indicated.

A. SDS-PAGE
1. Prepare 15% SDS-PAGE gel.
2. Prepare samples in microfuge tubes. Add 4X SDS sample buffer so the total protein amount is 25 μg per sample
3. Flick microfuge tubes to mix samples, and then heat to 95-100°C for 5 minutes.
4. Set up electrophoresis apparatus and immerse in 1X electrophoretic buffer.
Remove gel combs and cleanse wells of any residual stacking gel.
5. Load samples and protein markers onto the gel using gel loading tips. Set
electrophoresis power pack to 80V (through the stacking gel). Increase it to 120V
when the protein front reaches the separation gel.

B. Protein Transfer
1. Wet transfers are performed under 200mA constant current for 40min with an ice pack and at 4°C to mitigate the heat produced.
2. Sequentially assemble the transfer constituents and ensure no bubbles lie between any of the layers. Apply wet transfer systems according to the manufacturer’s
instructions.

C. Immunoblotting 2. After transfer, wash the membrane twice with distilled water, and using a pencil, mark bands of the MW ladder on the membrane.
3. Block with 1X TBST containing 3% nonfat dry milk with constant rocking for 1 hour .
4. Dilute primary antibody in blocking solution with a starting dilution ratio of 1:500.Incubate the membrane with primary antibody overnight at 4°C, on a bench-top rocker.
5. Wash membrane three times with 1X TBST for 10 minutes each.
6. Incubate the membrane with a suitable HRP-conjugated secondary antibody,diluted in 1X TBST. Incubate for 1 hour with constant rocking.
7. Wash membrane three times with 1X TBST for 10 minutes each.

D. Signal Detection
1. Prepare ECL substrate according to the manufacturer’s instructions.
2. Incubate the membrane completely with substrate for 20 seconds (adjust time for more sensitive ECL substrates).
3. Expose the membrane to autoradiography film in a dark room or read using a chemiluminescence imaging system.