Fresh savings just popped up! 25% off 3 or more products. | Start Saving >>
64459
Autophagy Atg8 Family Antibody Sampler Kit

Autophagy Atg8 Family Antibody Sampler Kit #64459

Western Blotting Image 1

Western blot analysis of extracts from HeLa and NIH/3T3 cells, untreated or chloroquine-treated (50 μM, overnight), using LC3A (D50G8) XP® Rabbit mAb.

Learn more about how we get our images
Western Blotting Image 2

Western blot analysis of extracts from various cell lines, untreated (-) or treated overnight with chloroquine (50 μM) (+), using LC3B (D11) XP® Rabbit mAb (upper) or LC3B Antibody #2775 (lower).

Learn more about how we get our images
Western Blotting Image 3

Western blot analysis of extracts from A172 and HOP-92 cells, untreated (-) or chloroquine-treated (50 μM, overnight; +), using LC3C (D3O6P) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Learn more about how we get our images
Western Blotting Image 4

Western blot analysis of A172 and C2C12 cells, untreated (-) or chloroquine-treated (50 μM, overnight; +), using GABARAP (E1J4E) Rabbit mAb.

Learn more about how we get our images
Western Blotting Image 5

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human GABARAP protein (hGABARAP-Myc/DDK; +), human GABARAPL1 protein (hGABARAPL1-Myc/DDK; +), or human GABARAPL2 protein (hGABARAPL2-Myc/DDK; +) using GABARAPL1 (D5R9Y) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Learn more about how we get our images
Western Blotting Image 6

Western blot analysis of extracts from 293T cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® GABARAPL2 siRNA I # 14239 (+) or SignalSilence® II GABARAPL2 siRNA II #14246 (+), using GABARAPL2 (D1W9T) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The GABARAPL2 (D1W9T) Rabbit mAb confirms silencing of GABARAPL2, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.

Learn more about how we get our images
Western Blotting Image 7

After the primary antibody is bound to the target protein, a complex with HRP-linked secondary antibody is formed. The LumiGLO® is added and emits light during enzyme catalyzed decomposition.

Learn more about how we get our images
IHC-P (paraffin) Image 8

Immunohistochemical analysis of paraffin-embedded human glioblastoma multiforme using LC3A (D50G8) XP® Rabbit mAb.

Learn more about how we get our images
Western Blotting Image 9

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® LC3B siRNA I #6212 (+) or SignalSilence® LC3B siRNA II #6213 (+), using LC3B (D11) XP® Rabbit mAb #3868 and α-Tubulin (11H10) Rabbit mAb #2125. The LC3B (D11) XP® Rabbit mAb confirms silencing of LC3B expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of LC3B siRNA.

Learn more about how we get our images
Western Blotting Image 10

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing full-length human LC3A protein (hLC3A; +), human LC3B protein (hLC3B; +), or human LC3C protein (hLC3C; +), using LC3C (D3O6P) Rabbit mAb.

Learn more about how we get our images
Western Blotting Image 11

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human GABARAP (hGABARAP-Myc/DDK; +), human GABARAPL1 (hGABARAPL1-Myc/DDK; +), or human GABARAPL2 (hGABARAPL2-Myc/DDK; +), using GABARAP (E1J4E) Rabbit mAb.

Learn more about how we get our images
Western Blotting Image 12

Western blot analysis of extracts from HeLa, C2C12, and C6 cells, untreated (-) or treated with chloroquine (50 μM, overnight; +), using GABARAPL1 (D5R9Y) XP® Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Learn more about how we get our images
Western Blotting Image 13

Western blot analysis of extracts from various cell lines using GABARAPL2 (D1W9T) Rabbit mAb.

Learn more about how we get our images
IHC-P (paraffin) Image 14

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using LC3A (D50G8) XP® Rabbit mAb (left) or

Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (right).

Learn more about how we get our images
IHC-P (paraffin) Image 15

Immunohistochemical analysis of paraffin-embedded human astrocytoma using LC3B (D11) XP® Rabbit mAb in the presence of control peptide (left) or antigen-specific peptide (right).

Learn more about how we get our images
IF-IC Image 16

Confocal immunofluorescent analysis of A172 cells, untreated (left) and chloroquine-treated (50 μM, overnight; right), using GABARAP (E1J4E) Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Learn more about how we get our images
Flow Cytometry Image 17

Flow cytometric analysis of HeLa cells using GABARAPL1 (D5R9Y) XP® Rabbit mAb (blue) compared to concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red). Anti-rabbit IgG (H+L), F(ab')2 Fragment (Alexa Fluor® 488 Conjugate) #4412 was used as a secondary antibody.

Learn more about how we get our images
Western Blotting Image 18

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with constructs expressing Myc/DDK-tagged full-length human GABARAP (hGABARAP-Myc/DDK; +), human GABARAPL1 (hGABARAPL1-Myc/DDK; +), or human GABARAPL2 (hGABARAPL2-Myc/DDK; +), using GABARAPL2 (D1W9T) Rabbit mAb.

Learn more about how we get our images
IHC-P (paraffin) Image 19

Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, control (left) or choloroquine-treated (right), using LC3A (D50G8) XP® Rabbit mAb.

Learn more about how we get our images
IHC-P (paraffin) Image 20

Immunohistochemical analysis of paraffin-embedded HeLa cell pellets, control (left) or chloroquine-treated (right), using LC3B (D11) XP® Rabbit mAb.

Learn more about how we get our images
IF-IC Image 21

Confocal immunofluorescent analysis of HeLa cells, untreated (left) or treated with chloroquine (50 μM, 24 hr; right), using GABARAPL1 (D5R9Y) XP® Rabbit mAb (green) and β-Actin (8H10D10) Mouse mAb #3700 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Learn more about how we get our images
Western Blotting Image 22

Western blot analysis of extracts from A172 and HeLa cells, untreated (-) or treated overnight with chloroquine (50 μM) (+), using GABARAPL2 (D1W9T) Rabbit mAb (upper) or β-Actin (D6A8) #8457 (lower).

Learn more about how we get our images
Flow Cytometry Image 23

Flow cytometric analysis of HeLa cells using LC3B (D11) XP® Rabbit mAb (blue) compared to a nonspecific negative control antibody (red).

Learn more about how we get our images
IF-IC Image 24

Confocal immunofluorescent analysis of HCT-116 cells, untreated (left) or choroquine-treated (50 uM, overnight; right) using LC3B (D11) XP® Rabbit mAb (green) and β-Catenin (L54E2) Mouse mAb (Alexa Fluor® 555 Conjugate) #5612 (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Learn more about how we get our images
Product Includes Quantity Applications Reactivity MW(kDa) Isotype
LC3A (D50G8) XP® Rabbit mAb 4599 20 µl
  • WB
  • IHC
H M R 14, 16 Rabbit IgG
LC3B (D11) XP® Rabbit mAb 3868 20 µl
  • WB
  • IP
  • IHC
  • IF
  • F
H M R 14, 16 Rabbit IgG
LC3C (D3O6P) Rabbit mAb 14736 20 µl
  • WB
H 14 Rabbit IgG
GABARAP (E1J4E) Rabbit mAb 13733 20 µl
  • WB
  • IF
H M R 14, 16 Rabbit IgG
GABARAPL1 (D5R9Y) XP® Rabbit mAb 26632 20 µl
  • WB
  • IF
  • F
H M R 14, 16 Rabbit IgG
GABARAPL2 (D1W9T) Rabbit mAb 14256 20 µl
  • WB
H M R Mk 14 Rabbit IgG
Anti-rabbit IgG, HRP-linked Antibody 7074 100 µl
  • WB
Goat 

The Autophagy Atg8 Family Antibody Sampler Kit provides an economical means of detecting each of the Atg8 family members. The kit contains enough primary antibody to perform at least two western blot experiments.

Each antibody in the Autophagy Atg8 Family Antibody Sampler Kit detects endogenous levels of its target protein. Cross-reactivity was not observed between family members.

Monoclonal antibodies are produced by immunizing rabbits with synthetic peptides corresponding to the amino terminus of human LC3A, LC3B, LC3C, and GABARAPL1, Arg40 of human GABARAP, and the carboxy terminus of GABARAPL2.

Autophagy is a catabolic process for the autophagosomic-lysosomal degradation of bulk cytoplasmic contents (1,2). Autophagy is generally activated by conditions of nutrient deprivation, but it has also been associated with a number of physiological processes, including development, differentiation, neurodegenerative diseases, infection, and cancer (3).

Atg8 is a ubiquitin-like protein that is critical for autophagosome formation. Atg8 is synthesized as a precursor protein that is processed by the cysteine protease Atg4, followed by lipidation with phosphatidylethanolamine (PE) in a ubiqutin-like conjugation pathway involving Atg7 and Atg3 (4). This processing of Atg8, which is described as a conversion from type-I to type-II forms, is frequently described as a marker for autophagy. The type-II form of Atg8 is incorporated into maturing autophagosomes and leads to the recruitment of additional autophagy components, including cargo receptors like SQSTM1/p62. While yeast has a single Atg8 gene, many eukaryotes have at least six orthologs, including three microtubule-associated protein 1 light chain 3 (MAP1LC3/LC3) family members (LC3A, LC3B, and LC3C) and three GABAA receptor associated protein (GABARAP) family members (GABARAP, GABARAPL1/GEC1, and GABARAPL2/GATE-16). While highly conserved, these various family members can have important differences in their post-translational processing, expression profile, and protein interactions including distinct cargo receptor. This complexity within the Atg8 family is critical for selective mechanisms of autophagy that have been reported (5, 6).

  1. Reggiori, F. and Klionsky, D.J. (2002) Eukaryot. Cell 1, 11-21.
  2. Codogno, P. and Meijer, A.J. (2005) Cell Death Differ. 12 Suppl 2, 1509-18.
  3. Levine, B. and Yuan, J. (2005) J. Clin. Invest. 115, 2679-88.
  4. Ichimura, Y. et al. (2000) Nature 408, 488-92.
  5. Slobodkin, M.R. and Elazar, Z. (2013) Essays Biochem 55, 51-64.
  6. Schaaf, M.B. et al. (2016) FASEB J 30, 3961-3978.
Entrez-Gene Id
11337 , 23710 , 11345 , 84557 , 81631 , 440738
Swiss-Prot Acc.
O95166 , Q9H0R8 , P60520 , Q9H492 , Q9GZQ8 , Q9BXW4
For Research Use Only. Not For Use In Diagnostic Procedures.

Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
XP is a registered trademark of Cell Signaling Technology, Inc.

Upstream / Downstream

pathwayImage

Explore pathways related to this product.