Name: Anti-BRAF antibody [EP152Y]
SKU: ab33899
Rating: 4 (12 reviews)

Anti-BRAF antibody [EP152Y] (ab33899) is a rabbit monoclonal antibody that is used to detect BRAF in Western Blot, IP, IHC-P. Suitable for Human, Mouse, Rat samples.

- Specificity confirmed with BRAF knockout cell line validation

Images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRAF antibody [EP152Y] (AB33899), expandable thumbnail

Publications

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal

Immunogen

The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application

Product promiseTestedExpectedPredictedNot recommended

	IP	WB	IHC-P	ICC
Human	Tested	Tested	Tested	Not recommended
Mouse	Expected	Tested	Expected	Not recommended
Rat	Expected	Tested	Expected	Not recommended

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/20	Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info 1/1000 - 1/5000	Notes -
Species Rat	Dilution info 1/1000 - 1/5000	Notes -
Species Human	Dilution info 1/1000 - 1/5000	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Human	Dilution info 1/100	Notes The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat. Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Expected
Expected

Species	Dilution info	Notes
Species Mouse, Rat	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Mouse, Rat, Human	Dilution info -	Notes -

Associated Products

Select an associated product type

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Target data

See full target information BRAF

Function

Protein kinase involved in the transduction of mitogenic signals from the cell membrane to the nucleus (Probable). Phosphorylates MAP2K1, and thereby activates the MAP kinase signal transduction pathway (PubMed:21441910, PubMed:29433126). Phosphorylates PFKFB2 (PubMed:36402789). May play a role in the postsynaptic responses of hippocampal neurons (PubMed:1508179).

Alternative names

BRAF1, RAFB1, BRAF, Serine/threonine-protein kinase B-raf, Proto-oncogene B-Raf, p94, v-Raf murine sarcoma viral oncogene homolog B1

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Clone number: EP152Y
Purification technique: Affinity purification Protein A
Specificity: The mouse and rat recommendation is based on the WB results. We do not guarantee IHC-P for mouse and rat.
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage duration: 1-2 weeks
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: -20°C
Aliquoting information: Upon delivery aliquot
Storage information: Avoid freeze / thaw cycle

Notes

What is this antibody validated in?

Anti-BRAF antibody [EP152Y] (ab33899) is a rabbit recombinant monoclonal antibody and is validated for use in Western Blot (WB), Immunoprecipitation (IP), Immunohistochemistry (IHC-P), in Human, Mouse, Rat samples.

What is the molecular weight of BRAF?

Anti-BRAF [EP152Y] (ab33899) specifically detects a band for BRAF (UniProt: P15056) at a molecular weight of 85kDa.

Recommended positive controls

WB: HAP1, PC-12 and HeLa whole cell lysate (ab150035); Mouse brain tissue lysate.IHC-P: Human prostate and breast cancer tissue; Rat brain tissue.IP: HeLa cells.

Trusted by the scientific community

Anti-BRAF [EP152Y] (ab33899) was first used in a scientific publication in 2006 and has been cited over 30 times in peer-reviewed journals.

Reviewed by scientists

Anti-BRAF [EP152Y] (ab33899) has over 10 independent reviews from customers.

Trial sizes available!

Test your antibody or perform pre-screening before committing to a larger quantity. Sold in 10µl. Discover our selection of trial-size antibodies. Specificity confirmed

The specificity of Anti-BRAF antibody [EP152Y] (ab33899) has been confirmed by Western blot testing in BRAF Knockout HAP1 cells.

Other related products

We have a range of other formats of antibody clone [EP152Y] also available for your convenience:
ab33899, Carrier free - ab189351, Alexa Fluor® 647 - ab309081, Alexa Fluor® 488 - ab309244

Patented technology
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

What are the advantages of a recombinant monoclonal antibody?
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 batch production

For more information, read more on recombinant antibodies.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

13 product images

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRAF antibody [EP152Y] (ab33899)
ab33899 (unpurified) staining B Raf cells from human prostate tissue by immunohistochemistry (formalin/PFA-fixed paraffin-embedded sections). Cells were formaldehyde fixed and permeabilized in PBS-Tween 20 prior to blocking in 70% serum for 10 minutes at 25°C. The primary antibody was diluted 1/250 and incubated with the sample for 1 hour at 25°C. A biotin conjugated goat polyclonal to mouse Ig was used as the secondary.
Western blot - Anti-BRAF antibody [EP152Y] (ab33899)
Lanes 1- 2: Merged signal (red and green). Green - ab33899 observed at 90 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) observed at 37 kDa.
ab33899 was shown to react with BRAF in wild-type HeLa cells in western blot. Loss of signal was observed when knockout cell line Human BRAF knockout HeLa cell line ab265373 (knockout cell lysate Human BRAF knockout HeLa cell lysate ab257078) was used. Wild-type HeLa and BRAF knockout HeLa cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab33899 and Anti-GAPDH antibody [6C5] - Loading Control (Anti-GAPDH antibody [6C5] - Loading Control ab8245) 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) preadsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye^®680RD) preadsorbed (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.
All lanes: Western blot - Anti-BRAF antibody [EP152Y] (ab33899) at 1/1000 dilution
Lane 1: Wild-type HeLa cell lysate at 20 µg
Lane 2: BRAF knockout HeLa cell lysate at 20 µg
Performed under reducing conditions.
Predicted band size: 84 kDa
Observed band size: 90 kDa
Immunoprecipitation - Anti-BRAF antibody [EP152Y] (ab33899)
ab33899 (Purified) at 1/20 dilution (20 μg/ml) immunoprecipitating BRAF in HeLa whole cell lysate.
Lane 1 (input): HeLa (human cervix adenocarcinoma epithelial cell) whole cell lysate 10 μg
Lane 2 (+): ab33899 & HeLa whole cell lysate
Lane 3 (-): Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab33899 in HeLa whole cell lysate
For western blotting, ab33899 at 1/500 and VeriBlot for IP secondary antibody (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) was used as the secondary antibody at 1/1000 dilution.
Blocking and diluting buffer: 5% NFDM /TBST .
All lanes: Immunoprecipitation - Anti-BRAF antibody [EP152Y] (ab33899)
Predicted band size: 84 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRAF antibody [EP152Y] (ab33899)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human breast cancer tissue sections labeling BRAF with purified ab33899 at 1/100 dilution (4.77 μg/ml). Heat mediated antigen retrieval was performed using heat mediated antigen retrieval using Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) ab93684 (Tris/EDTA buffer, pH 9.0). ImmunoHistoProbe one step HRP Polymer (ready to use) was used as the secondary antibody. Negative control: PBS instead of the primary antibody. Hematoxylin was used as a counterstain.
Western blot - Anti-BRAF antibody [EP152Y] (ab33899)
Lanes 1 - 3: Merged signal (red and green). Green - ab33899 (unpurified) observed at 90 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
ab33899 was shown to recognize B Raf in wild-type HAP1 cells as signal was lost at the expected MW in B Raf knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and B Raf knockout samples were subjected to SDS-PAGE. ab33899 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-BRAF antibody [EP152Y] (ab33899) at 1/1000 dilution
Lane 1: Wild-type HAP1 whole cell lysate at 40 µg
Lane 2: BRAF knockout HAP1 whole cell lysate at 40 µg
Lane 3: HeLa whole cell lysate at 40 µg
Predicted band size: 84 kDa
Western blot - Anti-BRAF antibody [EP152Y] (ab33899)
All lanes: Western blot - Anti-BRAF antibody [EP152Y] (ab33899) at 1/5000 dilution
All lanes: PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysates at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 84 kDa
Observed band size: 87 kDa
Western blot - Anti-BRAF antibody [EP152Y] (ab33899)
All lanes: Western blot - Anti-BRAF antibody [EP152Y] (ab33899) at 1/2000 dilution
Lane 1: HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysates at 15 µg
Lane 2: Mouse brain lysates at 15 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 84 kDa
Observed band size: 87 kDa
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-BRAF antibody [EP152Y] (ab33899)
This image shows paraffin embedded human prostate cancer tissue sample stained with ab33899 (unpurified) at 1/250 dilution.
Western blot - Anti-BRAF antibody [EP152Y] (ab33899)
All lanes: Western blot - Anti-BRAF antibody [EP152Y] (ab33899) at 1/5000 dilution
All lanes: HeLa cell lysate
Predicted band size: 84 kDa
Observed band size: 87 kDa
Western blot - Anti-BRAF antibody [EP152Y] (ab33899)
All lanes: Western blot - Anti-BRAF antibody [EP152Y] (ab33899) at 1/1000 dilution
Lane 1: Lysate prepared from mouse brain
Lane 2: Lysate prepared from mouse heart
Lane 3: Lysate prepared from mouse kidney
Lane 4: Lysate prepared from mouse spleen
Lane 5: Lysate prepared from rat brain
Lane 6: Lysate prepared from rat heart
Lane 7: Lysate prepared from rat kidney
Lane 8: Lysate prepared from rat spleen
Predicted band size: 84 kDa
Exposure time: 3min
Western blot - Anti-BRAF antibody [EP152Y] (ab33899)
Blocking and diluting buffer and concentration: 5% NFDM/TBST
The band below 90 kDa in lane 1 may be caused by degradation.
Lysates were freshly made and used immediately to minimize protein degradation.
In Western blot, anti-GAPDH antibody (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) loading control staining at 1/200000 dilution.
Anti-BRAF antibody (ab33899) total protein control staining at 1/1000 dilution.
Exposure time: Lane 1, 2: 3 minutes; Lane 3, 4: 15 seconds
All lanes: Western blot - Anti-BRAF (phospho S446) antibody [EPR28069-63] (Anti-BRAF (phospho S446) antibody [EPR28069-63] ab313793) at 1/1000 dilution
Lane 1: NIH/3T3 (mouse embryonic fibroblast) whole cell lysate (untreated membrane) at 20 µg
Lane 2: NIH/3T3 whole cell lysate (phosphatase treated membrane) at 20 µg
Lane 3: PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate (untreated membrane) at 20 µg
Lane 4: PC-12 whole cell lysate (phosphatase treated membrane) at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 90 kDa
Western blot - Anti-BRAF antibody [EP152Y] (ab33899)
BRAF Western blot staining using rabbit Anti-BRAF antibody
Blocking and diluting buffer and concentration: 5% NFDM/TBST
The bands below 90 kDa in lane 1 may be caused by degradation. Lysates were freshly made and used immediately to minimize protein degradation.
In Western blot, anti- Vinculin antibody (Anti-Vinculin antibody [EPR8185] - Loading Control ab129002) loading control staining at 1/10000 dilution.
Anti-BRAF antibody (ab33899) total protein control staining at 1/1000 dilution.
Exposure time: 3 minutes
All lanes: Western blot - Anti-BRAF (phospho S446) antibody [EPR28069-63] (Anti-BRAF (phospho S446) antibody [EPR28069-63] ab313793) at 1/1000 dilution
Lane 1: Wild-type HAP1(human chronic myelogenous leukemia near-haploid cell line) whole cell lysate (untreated membrane) at 20 µg
Lane 2: BRAF knockout HAP1 whole cell lysate (untreated membrane) at 20 µg
Lane 3: Wild-type HAP1 whole cell lysate (phosphatase treated membrane) at 20 µg
Lane 4: BRAF knockout HAP1 whole cell lysate (phosphatase treated membrane) at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Observed band size: 90 kDa
Exposure time: 3min
Image collected and cropped by CiteAb under a CC-BY license from the publication
Western blot - Anti-BRAF antibody [EP152Y] (ab33899)
BRAF western blot using anti-BRAF antibody [EP152Y] ab33899. Publication image and figure legend from Zhang, H., Zheng, D., et al., 2016, Sci Rep, PubMed 27767059.

ab33899 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab33899 please see the product overview.
Compounds 1, 2, and 9 downregulate RAF-MAPK signaling pathway and snail protein level.RAS-RAF-MEK-ERK cascades and snail protein were analyzed by western blot. TE1 cells were treated with 1, 2, or 9 with various concentrations (0, 10, 20 μM) for 48 h. Sorafenib (SFB) was used as the positive control.