Antibodies, Proteins, Kits and Reagents for Life Science

304 886 ₸ Product size

100 µl 304 886 ₸

Order now and get it on Tuesday March 02, 2021

Anti-MAPK6/ERK3 antibody [EP1720Y] (ab53277)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP1720Y] to MAPK6/ERK3
Suitable for: WB, IHC-P
Knockout validated
Reacts with: Mouse, Rat, Human

Product name

Anti-MAPK6/ERK3 antibody [EP1720Y]
See all MAPK6/ERK3 primary antibodies
Description

Rabbit monoclonal [EP1720Y] to MAPK6/ERK3
Host species

Rabbit
Specificity

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.

Tested Applications & Species

Application	Species
IHC-P	Human
WB	Mouse Rat Human

See all applications and species data

Immunogen

Synthetic peptide within Human MAPK6/ERK3 aa 1-100 (N terminal). The exact sequence is proprietary.
Database link: Q16659
Positive control
- WB: Wild-type HAP1, A431, C2C12, 2.4G2 and HeLa (ab150035) whole cell lysates, and rat brain tissue lysates. IHC-P: Human kidney carcinoma and breast carcinoma tissues.
General notes
Previously labelled as MAPK6.

The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EP1720Y
Isotype

IgG
Research areas

Western blot - Anti-MAPK6/ERK3 antibody [EP1720Y] (ab53277)

All lanes : Anti-MAPK6/ERK3 antibody [EP1720Y] (ab53277) at 1/1000 dilution

Lane 1 : Wild-type HeLa cell lysate
Lane 2 : MAPK6 knockout HeLa cell lysate
Lane 3 : 2.4G2 cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

Predicted band size: 63 kDa
Observed band size: 90 kDa
why is the actual band size different from the predicted?

Lanes 1-3: Merged signal (red and green). Green - ab53277 observed at 90 kDa. Red - loading control ab8245 observed at 36 kDa.

ab53277 Anti-MAPK6/ERK3 antibody [EP1720Y] was shown to specifically react with MAPK6/ERK3 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab264910 (knockout cell lysate ab257526) was used. Wild-type and MAPK6/ERK3 knockout samples were subjected to SDS-PAGE. ab53277 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated at room temperature for 2.5 hours at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MAPK6/ERK3 antibody [EP1720Y] (ab53277)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human kidney carcinoma tissue sections labeling MAPK6/ERK3 with Purified ab53277 at 1:100 dilution (13.52 µg/ml). Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0). Purified)ImmunoHistoProbe one step HRP Polymer (ready to use) was used for detection. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Western blot - Anti-MAPK6/ERK3 antibody [EP1720Y] (ab53277)

All lanes : Anti-MAPK6/ERK3 antibody [EP1720Y] (ab53277) at 1/1000 dilution

Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : MAPK6 knockout HAP1 whole cell lysate
Lane 3 : A431 whole cell lysate
Lane 4 : MCF7 whole cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 63 kDa
Observed band size: 90 kDa why is the actual band size different from the predicted?

Lanes 1 - 4: Merged signal (red and green). Green - ab53277 observed at 90 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab53277 was shown to react with MAPK6 in HAP1 wild-type cells in Western blot. Loss of signal was observed when MAPK6 knockout sample was used. HAP1 wild-type and MAPK6 knockout whole cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween^®) before incubation with ab53277 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 800CW) preabsorbed (ab216772) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Western blot - Anti-MAPK6/ERK3 antibody [EP1720Y] (ab53277)

All lanes : Anti-MAPK6/ERK3 antibody [EP1720Y] (ab53277) at 1/1000 dilution (Purified)

Lane 1 : HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates
Lane 2 : Rat brain lysates
Lane 3 : C2C12 (Mouse myoblasts myoblast) whole cell lysates

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 63 kDa
Observed band size: 105 kDa why is the actual band size different from the predicted?

The molecular weight observed is consistent with what has been described in PMID:30642948 and 30166347
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-MAPK6/ERK3 antibody [EP1720Y] (ab53277)

Unpurified ab53277 (1:50) staining human MAPK6/ERK3 in human breast carcinoma tissue by immunohistochemistry using paraffin embedded tissue.

Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
Anti-MAPK6/ERK3 antibody [EP1720Y] (ab53277)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com