bsm-52259R [Primary Antibody]
ERK1/2 Recombinant Antibody
www.biossusa.com
[email protected]
800.501.7654 [DOMESTIC]
+1.781.569.5821 [INTERNATIONAL]
DATASHEET

Host: Rabbit

Target Protein: ERK1/2

Clonality: Recombinant

Isotype: IgG

Swiss Prot: P27361, P28482

Source: Recombinant Human ERK1 + ERK2 between 150 amino acids to C-terminus

Purification: Purified by Protein A.

Storage Buffer: 0.01M TBS(pH7.4) with 1% BSA, 0.02% Proclin300 and 50% Glycerol.

Storage: Store at -20°C for 12 months.

Background:

p44/42 MAP Kinase(Thr202); ERK (extracellular signal regulated kinase), also known as MAPK (mitogen activated protein kinase) has two closely related isoforms of 44 kDa and 42 kDa, respectively. These kinases belong to a family of serine/threonine kinases that are activated upon treatment of cells with a large variety of stimuli including mitogens, hormones, growth factors, cytokines, and bioactive peptides. Cell stimulation induces the activation of a signaling cascade, the downstream effects of which have been linked to the regulation of cell growth and differentiation as well as the cytoskeleton. ERK1 and ERK2 are phosphorylated within the activation loop on both a Threonine and a Tyrosine residue (within a Thr-Glu-Tyr motif) by MEKs (MAPK/ERK kinases), thereby greatly elevating the activity of ERK1&2.

Size: 100ul

Concentration: Lot dependent

Cross Reactive Species: Human
Mouse
Rat
Zebrafish

For research use only. Not intended for diagnostic or therapeutic use.

PRODUCT SPECIFIC PUBLICATIONS
  • Yudan Zhao. et al. COX-2 is required to mediate crosstalk of ROS-dependent activation of MAPK/NF-_B signaling with pro-inflammatory response and defense-related NO enhancement during challenge of macrophage-like cell line with Giardia duodenalis. PLOS NEGLECT TROP D. 2022 Apr;16(4):e0010402Read more>>
  • Jia Li. et al. Study of the Mechanism of Antiemetic Effect of <em>Lavandula angustifolia</em> Mill. Essential Oil Based on Ca<sup>2+</sup>/CaMKII/ERK1/2 Pathway. DRUG DES DEV THER. 2022 Jul;16:2407-2422Read more>>
  • Xing-Peng Di. et al. YAP/Smad3 promotes pathological extracellular matrix microenviroment-induced bladder smooth muscle proliferation in bladder fibrosis progression. MedComm. 2022 Sep;3(4):e169Read more>>
  • Honghong Zhan. et al. Oxybaphus himalaicus Mitigates Lipopolysaccharide-Induced Acute Kidney Injury by Inhibiting TLR4/MD2 Complex Formation. ANTIOXIDANTS-BASEL. 2022 Dec;11(12):2307Read more>>
  • Yiming Bi. et al. -Sitosterol Suppresses LPS-Induced Cytokine Production in Human Umbilical Vein Endothelial Cells via MAPKs and NF-<i></i>B Signaling Pathway. EVID-BASED COMPL ALT. 2023 Jan 03;2023:924109Read more>>
  • Lu Yi. et al. The protective role of chicken cathelicidin-1 against Streptococcus suis serotype 2 in vitro and in vivo. VET RES. 2023 Dec;54(1):1-13Read more>>
  • Myeongjoo Son. et al. Olive Flounder By-Product Prozyme2000P Hydrolysate Ameliorates Age-Related Kidney Decline by Inhibiting Ferroptosis. INT J MOL SCI. 2024 Jan;25(9):4668Read more>>
VALIDATION IMAGES

Lane 1: Hela; Lane 2: SW480; Lane 3: HCT116; Lane 4: PC12 lysate probed with ERK1/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:1000 overnight at 4°C followed by a conjugated secondary antibody for 60 minutes at 37°C.


Flow cytometric analysis of HeLa cells with ERK1/2 (3A12) Monoclonal Antibody (bsm-52259R) at a 1:50 dilution (red) compared with an unlabeled control (cells without incubation with primary antibody; black).


Flow cytometric analysis of SH-SY-5Y cells with ERK1/2 antibody at 1/50 dilution (blue) compared with an unlabelled control (cells without incubation with primary antibody; red). Alexa Fluor 488-conjugated goat anti-rabbit IgG was used as the secondary antibody.


IF(ICC) staining with ERK1/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:100 in HeLa cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.


IF(ICC) staining with ERK1/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:100 in MCF-7 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.


IF(ICC) staining with ERK1/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:100 in NIH/3T3 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.


IF(ICC) staining with ERK1/2 (3A12) Monoclonal Antibody (bsm-52259R) at 1:100 in A549 cells (green). The nuclear counterstain is DAPI (blue). Cells were fixed in paraformaldehyde, permeabilized with 0.25% Triton X100/PBS.


Lane 1: Mouse Cerebrum lysates; Lane 2: Mouse Pancreas lysates; Lane 3: Mouse Large intestine lysates; Lane 4: Mouse Lymph node lysates; Lane 5: Rat Cerebrum lysates; Lane 6: Rat Pancreas lysates; Lane 7: Rat Large intestine lysates; Lane 8: Rat Lymph node lysates; Lane 9: Human Hela cell lysates; Lane 10: Human SW480 cell lysates; Lane 11: Human MCF-7 cell lysates; Lane 12: Mouse NIH/3T3 cell lysates; Lane 13: Human A549 cell lysates; Lane 14: Human A431 cell lysates probed with ERK1/2 (3A12) Monoclonal Antibody, Unconjugated (bsm-52259R) at 1:1000 dilution and 4˚C overnight incubation. Followed by conjugated secondary antibody incubation at 1:20000 for 60 min at 37˚C.


Hela cells were fixed with 4% PFA for 10min at room temperature,permeabilized with 90% ice-cold methanol for 20 min at -20℃, and incubated in 5% BSA blocking buffer for 30 min at room temperature. Cells were then stained with ERK1/2 (3A12) Monoclonal Antibody(bs-R)at 1:100 dilution in blocking buffer and incubated for 30 min at room temperature, washed twice with 2%BSA in PBS, followed by secondary antibody incubation for 40 min at room temperature. Acquisitions of 20,000 events were performed. Cells stained with primary antibody (green), and isotype control (orange).


Tissue/cell:A549 cell;4% Paraformaldehyde-fixed;Triton X-100 at room temperature for 20 min; Blocking buffer (normal goat serum,C-0005) at 37°C for 20 min; Antibody incubation with (ERK1/2) monoclonal Antibody, Unconjugated (bsm-52259R) 1:100, 90 minutes at 37°C; followed by a FITC conjugated Goat Anti-Rabbit IgG antibody at 37°C for 90 minutes, DAPI (blue, C02-04002) was used to stain the cell nuclei.


Paraformaldehyde-fixed, paraffin embedded Rat colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with ERK1/2 (3A12) Monoclonal Antibody, Unconjugated (bsm-52259R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Rat brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with ERK1/2 (3A12) Monoclonal Antibody, Unconjugated (bsm-52259R) at 1:200 overnight at 4°C, DAB staining.


Paraformaldehyde-fixed, paraffin embedded Mouse brain; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15min; Block endogenous peroxidase by 3% hydrogen peroxide for 20 minutes; Blocking buffer (normal goat serum) at 37°C for 30min; Antibody incubation with ERK1/2 (3A12) Monoclonal Antibody, Unconjugated (bsm-52259R) at 1:200 overnight at 4°C, DAB staining.


25 ug total protein per lane of various lysates (see on figure) probed with ERK1/2 monoclonal antibody, unconjugated (bsm-52259R) at 1:1000 dilution and 4°C overnight incubation. Followed by conjugated secondary antibody incubation at r.t. for 60 min.


Paraformaldehyde-fixed, paraffin embedded Mouse Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human Colon; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human Colon Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human Cervical Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human Breast Cancer; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human Tonsil; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human Cerebrum; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human Breast; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.


Paraformaldehyde-fixed, paraffin embedded Human Placenta; Antigen retrieval by boiling in sodium citrate buffer (pH6.0) for 15 min; Antibody incubation with ERK1/2 Monoclonal Antibody, Unconjugated(bsm-52259R) at 1:250 overnight at 4°C, followed by conjugation to the SP Kit (Rabbit, SP-0023) and DAB (C-0010) staining.