ER Stress Antibody Sampler Kit
cat.: HAK21037
Product Type: Antibody Sampler Kit
Species reactivity: Human
Applications: WB
Form: Liquid
Storage condition: Store at +4℃ after thawing. Aliquot store at -20℃. Avoid repeated freeze / thaw cycles.
Storage buffer: 1*TBS (pH7.4), 0.1% BSA, 40% Glycerol. Preservative: 0.05% Sodium Azide.
Uniprot #: SwissProt: P11021 Human | P20029 Mouse | P06761 Rat | P27824 Human | P35564 Mouse | P35565 Rat | Q96HE7 Human | Q8R180 Mouse | Q8R4A1 Rat | O75460 Human | Q9EQY0 Mouse | P07237 Human | P09103 Mouse | P04785 Rat | P35638 Human | P35639 Mouse | Q62857 Rat | Q9NZJ5 Human
Entrez Gene: 498013 Rat
Images
HAK21037_1.jpg Fig1: Western blot analysis of GRP78 / BIP on different lysates with Mouse anti-GRP78 / BIP antibody (HA601076) at 1/1,000 dilution.

Lane 1: MCF7 cell lysate (15 µg/Lane)
Lane 2: HepG2 cell lysate (15 µg/Lane)
Lane 3: Mouse brain tissue lysate (30 µg/Lane)
Lane 4: U-87 MG cell lysate (30 µg/Lane)
Lane 5: RAW264.7 cell lysate (30 µg/Lane)
Lane 6: RAW264.7 treated with 300nM Thapsigargin for 18 hours cell lysate (30 µg/Lane)
Lane 7: Mouse liver tissue lysate (30 µg/Lane)
Lane 8: Rat liver tissue lysate (30 µg/Lane)
Lane 9: Rat pancreas tissue lysate (30 µg/Lane)

Predicted band size: 72 kDa
Observed band size: 72 kDa

Exposure time: Lane 1-3: 11 seconds; Lane 4-9: 4 seconds; ECL: K1801;

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 (HA601076) at 1/1,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Mouse IgG - HRP Secondary Antibody (HA1006) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21037_2.png Fig2: Immunocytochemistry analysis of Hela cells labeling GRP78 / BIP with Mouse anti-GRP78 / BIP antibody (HA601076) at 1/200 dilution.

Cells were fixed in 100% methanol for 10 minutes, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes, and then blocked with 1% BSA for 30 minutes at room temperature. Cells were then incubated with Mouse anti-GRP78 / BIP antibody (HA601076) at 1/200 dilution in 2% negative goat serum overnight at 4 ℃. Goat Anti-Mouse IgG H&L (iFluor™ 488, HA1125) was used as the secondary antibody at 1/1,000 dilution. Nuclear DNA was labelled in blue with DAPI.

beta Tubulin (ET1602-4, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Rabbit IgG H&L (iFluor™ 594, HA1122) were used as the secondary antibody at 1/1,000 dilution.
HAK21037_3.jpg Fig3: Western blot analysis of Calnexin on different lysates with Rabbit anti-Calnexin antibody (ET1611-86) at 1/2,000 dilution.

Lane 1: HeLa cell lysate
Lane 2: HepG2 cell lysate
Lane 3: MCF7 cell lysate
Lane 4: PANC-1 cell lysate

Lysates/proteins at 15 µg/Lane.

Predicted band size: 68 kDa
Observed band size: 100 kDa

Exposure time: 2 minutes;

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 (ET1611-86) at 1/2,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21037_4.jpg Fig4: Western blot analysis of DDIT3 on different lysates with Rabbit anti-DDIT3 antibody (ET1703-05) at 1/5,000 dilution.

Lane 1: SW480 cell lysate (20 µg/Lane)
Lane 2: HeLa cell lysate (20 µg/Lane)

Predicted band size: 19 kDa
Observed band size: 27 kDa

Exposure time: 3 minutes;

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 (ET1703-05) at 1/5,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature.
HAK21037_5.jpg Fig5: Western blot analysis of PERK on different lysates with Rabbit anti-PERK antibody (HA721510) at 1/1,000 dilution.

Lane 1: MCF7 cell lysate (20 µg/Lane)
Lane 2: HepG2 cell lysate (20 µg/Lane)
Lane 3: HEK-293 cell lysate (20 µg/Lane)
Lane 4: HeLa cell lysate (20 µg/Lane)
Lane 5: A549 cell lysate (20 µg/Lane)
Lane 6: U-87 MG cell lysate (20 µg/Lane)
Lane 7: K-562 cell lysate (16 µg/Lane)

Predicted band size: 125 kDa
Observed band size: 140 kDa

Exposure time: 2 minutes;

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 (HA721510) 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.
Note: All products are “FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE”.