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Microtubule-Associated Protein 1 Light Chain 3 alpha (MAP1LC3A) (AA 25-121), (Internal Region) antibody

Details for Product No. ABIN250723, Supplier: Log in to see
Antigen
  • 1010001H21Rik
  • 4922501H04Rik
  • Anx3
  • ATG8E
  • b3gnt5
  • fb12f12
  • Lc3
  • LC3
  • LC3a
  • LC3A
  • LRRGT00047
  • MAP1ALC3
  • MAP1BLC3
  • Map1lc3
  • map1lc3a
  • MAP1LC3A
  • MGC69006
  • MGC89867
  • wu:fb12f12
  • wu:fb60g11
  • wu:fe50g08
  • zgc:56434
  • zgc:77094
Epitope
AA 25-121, Internal Region
36
30
28
24
23
18
14
14
14
12
12
11
10
8
8
7
7
6
6
5
5
4
3
3
3
2
2
2
2
2
2
2
2
2
1
1
1
1
1
1
1
1
1
1
1
1
Reactivity
Dog (Canine), Fish, Human, Mouse (Murine), Rat (Rattus), Zebrafish (Danio rerio)
286
80
74
17
11
6
5
2
1
1
1
1
1
Host
Rabbit
249
31
5
4
Clonality
Polyclonal
Conjugate
Un-conjugated
23
19
18
18
13
13
2
2
2
1
1
1
1
1
1
1
1
1
Application
ELISA, Flow Cytometry (FACS), Immunocytochemistry (ICC), Immunofluorescence (IF), Immunohistochemistry (IHC), Immunohistochemistry (Frozen Sections) (IHC (fro)), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP), Simple Western (SimWes), Southern Blotting (SB), Western Blotting (WB)
227
144
112
61
36
27
17
14
14
12
10
6
3
Supplier
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Immunogen A synthetic peptide made to an internal portion of the human LC3 protein sequence (between residues 25-121). [UniProt# Q9GZQ8].
Specificity This antibody detects both LC3A and LC3B.
Cross-Reactivity (Details) Predicted to react with Xenopus,bovine based on 100% sequence homology.
Purification Immunogen affinity purified
Alternative Name LC3/MAP1LC3A (MAP1LC3A Antibody Abstract)
Background Gene Symbol: MAP1LC3A
UniProt Q9GZQ8
Application Notes Western Blot 2 μg/mL, Simple Western 1:50, Flow Cytometry, ELISA, Immunohistochemistry 1:200-1:400, Immunocytochemistry/Immunofluorescence 1:100-1:300, Immunoprecipitation 20 μg/ 500 μg of lysate, Immunohistochemistry-Paraffin 1:200-1:400, Immunohistochemistry-Frozen, Southern Blot
Comment

The antibodies are intended for use in vitro experiments only. Our antibodies have not been tested nor are recommended for use in vivo.

Protocol Western Blot protocol specific for LC3 Antibody Western Blot Protocol
1. Perform SDS-PAGE (4-12 %) on samples to be analyzed, loading 40 µg of total protein per lane.
. Transfer proteins to Nitrocellulose according to the instructions provided by the manufacturer of the transfer apparatus.
. Rinse membrane with dH2O and then stain the blot using ponceau S for 1-2 minutes to access the transfer of proteins onto the nitrocellulose membrane. Rinse the blot in water to remove excess stain and mark the lane locations and locations of molecular weight markers using a pencil.
. Rinse the blot in TBS for approximately 5 minutes.
. Block the membrane using 5 % non-fat dry milk + 1 % BSA in TBS for 1 hour at room temperature (RT).
. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1 % Tween] 3 times for 10 minutes each.
. Dilute the rabbit anti-LC3 primary antibody in blocking buffer and incubate 1 hour at RT.
. Rinse the membrane in dH2O and then wash the membrane in wash buffer [TBS + 0.1 % Tween] 3 times for 10 minutes each.
. Apply the diluted rabbit-IgG HRP-conjugated secondary antibody in blocking buffer (as per manufacturer's instructions) and incubate 1 hour at room temperature.
. Wash the blot in wash buffer [TBS + 0.1 % Tween] 3 times for 10 minutes each (this step can be repeated as required to reduce background).
. Apply the detection reagent of choice in accordance with the manufacturers instructions (we used BioFX Super Plus ECL). Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2 %, provided it does not interfere with antibody-antigen binding.I. Deparaffinization: A. Treat slides with Xylene: 3 changes for 5 minutes each. Drain slides for 10 seconds between changes.B. Treat slides with 100 % Reagent Alcohol: 3 changes for 5 minutes each. Drain slides for 10 seconds between changes.II. Quench Endogenous Peroxidase: To Prepare 200 mL of Quenching Solution: Add 3 mL of 30 % Hydrogen Peroxide to 200 mL of Methanol.**Use within 4 hours of preparationA.Place slides in peroxidase quenching solution: 15-30 minutes.III. Retrieve Epitopes: A. Preheat Citrate
Restrictions For Research Use only
Format Liquid
Concentration 1.0 mg/mL
Buffer PBS
Buffer contains: 0.02 % Sodium Azide
Preservative Sodium azide
Precaution of Use This product contains Sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Avoid freeze-thaw cycles
Storage -20 °C,-80 °C
Storage Comment Aliquot and store at -20°C or -80°C. Avoid freeze-thaw cycles.
Supplier Images
Western Blotting (WB) image for anti-Microtubule-Associated Protein 1 Light Chain 3 alpha (MAP1LC3A) (AA 25-121), (Internal Region) antibody (ABIN250723) anti-Microtubule-Associated Protein 1 Light Chain 3 alpha (MAP1LC3A) (AA 25-121), (Internal Region) antibody
Immunohistochemistry (IHC) image for anti-Microtubule-Associated Protein 1 Light Chain 3 alpha (MAP1LC3A) (AA 25-121), (Internal Region) antibody (ABIN250723) Staining of human brain, cerebral cortex, cell crocesses in gray matter using ABIN250...
Product cited in: Lee, Finkel: "Regulation of autophagy by the p300 acetyltransferase." in: The Journal of biological chemistry, Vol. 284, Issue 10, pp. 6322-8, 2009

Lee, Cao, Mostoslavsky, Lombard, Liu, Bruns, Tsokos, Alt, Finkel: "A role for the NAD-dependent deacetylase Sirt1 in the regulation of autophagy." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 105, Issue 9, pp. 3374-9, 2008

Swerdlow, McColl, Rong, Lam, Gupta, Distelhorst: "Apoptosis inhibition by Bcl-2 gives way to autophagy in glucocorticoid-treated lymphocytes." in: Autophagy, Vol. 4, Issue 5, pp. 612-20, 2008

Zhang, Yu, Pan, Hu, Hao, Cai, Zhu, Yu, Xie, Ma, Yuan: "Small molecule regulators of autophagy identified by an image-based high-throughput screen." in: Proceedings of the National Academy of Sciences of the United States of America, Vol. 104, Issue 48, pp. 19023-8, 2007

Kabeya, Mizushima, Ueno, Yamamoto, Kirisako, Noda, Kominami, Ohsumi, Yoshimori: "LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing." in: The EMBO journal, Vol. 19, Issue 21, pp. 5720-8, 2000

Background publications Mizushima, Yamamoto, Hatano, Kobayashi, Kabeya, Suzuki, Tokuhisa, Ohsumi, Yoshimori: "Dissection of autophagosome formation using Apg5-deficient mouse embryonic stem cells." in: The Journal of cell biology, Vol. 152, Issue 4, pp. 657-68, 2001