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Antibodies

Rabbit Anti-Phospho-RIP3 (S232) Antibody [JE44-62], Recombinant Rabbit monoclonal Antibody

Product Sizes
100uL
HA721428-HA721428
About this Product
SKU:
HA721428
Application:
WB, IF-Cell
Buffer:
1*TBS (pH7.4), 0.05% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
translate.label.attr.clone:
JE44-62
Clonality:
Recombinant Rabbit monoclonal Antibody
Concentration:
1 mg/mL.
Control:
L-929 treated with 20μM Z-VAD for 3.5 hours add 100nM SM-164 and 20ng/ml TNF-α for 3 hours whole cel
Format:
Liquid
Host:
Rabbit
Immunogen:
Synthetic phospho-peptide corresponding to residues surrounding Ser232 of Human RIP3.
Isotype:
IgG
Molecular Weight:
Predicted band size: 53 kDa
Purification:
Protein A affinity purified.
Reactivities:
Mouse, Rat
Shipping Conditions:
Blue Ice
Storage Conditions:
Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Supplier:
Huabio
Western blot analysis of Phospho-RIP3 (S232) on different lysates with Rabbit anti-Phospho-RIP3 (S232) antibody (HA721428) at 1/1,000 dilution. Lane 1: L-929 whole cell lysate Lane 2: L-929 treated with 20μM Z-VAD for 3.5 hours, add 100nM SM-164 and 20ng/ml TNF-α for 3 hours whole cell lysate Lane 3: L-929 treated with λpp for 1 hour whole cell lysate Lane 4: L-929 treated with 20μM Z-VAD for 3.5 hours, add 100nM SM-164 and 20ng/ml TNF-α for 3 hours, treated with λpp for 1 hour whole cell lysate Lysates/proteins at 20 µg/Lane. Predicted band size: 53 kDa Observed band size: 53 kDa Exposure time: 20 seconds; 4-20% SDS-PAGE gel. Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA721428) at 1/1,000 dilution was used in 5% NFDM/TBST at room temperature for 2 hours. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1:100,000 dilution was used for 1 hour at room temperature.
Immunocytochemistry analysis of L-929 cells treated with or without 20μM Z-VAD for 2.75 hours, add 100nM SM-164 and 20ng/ml TNF-α for 2.25 hours, then treated with λpp for 1 hour labeling Phospho-RIP3 (S232) with Rabbit anti-Phospho-RIP3 (S232) antibody (HA721428) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 10 minutes at 37 ℃, permeabilized with 0.05% Triton X-100 in PBS for 20 minutes, and then blocked with 2% negative goat serum for 30 minutes at room temperature. Cells were then incubated with Rabbit anti-Phospho-RIP3 (S232) antibody (HA721428) at 1/100 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor[TM] 488, HA1121) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI. Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor[TM] 594, HA1126) was used as the secondary antibody at 1/1,000 dilution.