Anti-Egr1 antibody [EPR23981-46] (BSA and Azide free) (ab300450)

Rabbit Monoclonal EGR1 antibody. Carrier free. Suitable for IP, ICC/IF, IHC-Fr, IHC-P, WB and reacts with Rat, Mouse, Human samples.

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Images

Immunohistochemistry (Frozen sections) - Anti-Egr1 antibody [EPR23981-46] (BSA and Azide free) (AB300450), expandable thumbnail

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
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	Flow Cyt (Intra)	ICC/IF	IHC-Fr	IHC-P	WB
Human	Expected	Not recommended	Expected	Expected	Tested	Tested
Mouse	Tested	Not recommended	Tested	Tested	Tested	Tested
Rat	Tested	Not recommended	Tested	Not recommended	Tested	Tested

Tested
Tested

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

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

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

Tested
Tested

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

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Tested
Tested

Species	Dilution info	Notes
Species Mouse	Dilution info -	Notes -

Expected
Expected

Species	Dilution info	Notes
Species Human	Dilution info Use at an assay dependent concentration.	Notes -

Not recommended
Not recommended

Species	Dilution info	Notes
Species Rat	Dilution info -	Notes -

Tested
Tested

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

Tested
Tested

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

Target data

See full target information EGR1

Function

Transcriptional regulator (PubMed:20121949). Recognizes and binds to the DNA sequence 5'-GCG(T/G)GGGCG-3'(EGR-site) in the promoter region of target genes (By similarity). Binds double-stranded target DNA, irrespective of the cytosine methylation status (PubMed:25258363, PubMed:25999311). Regulates the transcription of numerous target genes, and thereby plays an important role in regulating the response to growth factors, DNA damage, and ischemia. Plays a role in the regulation of cell survival, proliferation and cell death. Activates expression of p53/TP53 and TGFB1, and thereby helps prevent tumor formation. Required for normal progress through mitosis and normal proliferation of hepatocytes after partial hepatectomy. Mediates responses to ischemia and hypoxia; regulates the expression of proteins such as IL1B and CXCL2 that are involved in inflammatory processes and development of tissue damage after ischemia. Regulates biosynthesis of luteinizing hormone (LHB) in the pituitary (By similarity). Regulates the amplitude of the expression rhythms of clock genes: BMAL1, PER2 and NR1D1 in the liver via the activation of PER1 (clock repressor) transcription. Regulates the rhythmic expression of core-clock gene BMAL1 in the suprachiasmatic nucleus (SCN) (By similarity).

Alternative names

KROX24, ZNF225, EGR1, Early growth response protein 1, EGR-1, AT225, Nerve growth factor-induced protein A, Transcription factor ETR103, Transcription factor Zif268, Zinc finger protein 225, Zinc finger protein Krox-24, NGFI-A

Recommended products

Rabbit Monoclonal EGR1 antibody. Carrier free. Suitable for IP, ICC/IF, IHC-Fr, IHC-P, WB and reacts with Rat, Mouse, Human samples.

Key facts

Isotype: IgG
Host species: Rabbit
Storage buffer: pH: 7.2 - 7.4
Constituents: 100% PBS
Form: Liquid
Clonality: Monoclonal
Immunogen: The exact immunogen used to generate this antibody is proprietary information.
Carrier free: Yes
Clone number: EPR23981-46
Purification technique: Affinity purification Protein A
Concentration

Storage

Shipped at conditions: Blue Ice
Appropriate short-term storage conditions: +4°C
Appropriate long-term storage conditions: +4°C

Notes

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

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.

Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.

This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.

Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

This product is compatible with the Maxpar^® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar^® is a trademark of Fluidigm Canada Inc.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Egr1 also known as early growth response protein 1 is a zinc-finger transcription factor with a molecular weight of approximately 60 kDa. The Egr1 protein plays a significant role in gene expression regulation by binding to specific DNA sequences. This target is highly expressed in a variety of tissues including the brain heart and kidneys. In these tissues Egr1 regulates several cellular processes by activating or repressing target genes. Also Egr1 has monoclonal antibodies available for research purposes providing tools for detailed study in specific cellular contexts.

Biological function summary

Egr1 acts as an immediate-early response gene. It gets rapidly upregulated in response to growth factors stress signals and other extracellular stimuli. As part of the cellular signaling networks Egr1 interacts with other transcription factors to control the expression of genes involved in cell differentiation proliferation and apoptosis. It does not typically form part of a protein complex but may work in concert with other signaling molecules to influence diverse cellular pathways and maintain cellular homeostasis.

Pathways

Egr1 is involved in the MAPK/ERK and PI3K/AKT signaling pathways both key to regulating cellular growth and survival. It functions as a bridging factor that links extracellular signals to transcriptional changes within the nucleus. In the MAPK/ERK pathway Egr1 works alongside proteins like Ras and MEK to promote early gene transcription in response to mitogenic signals. In the PI3K/AKT pathway Egr1 can influence downstream targets reinforcing the pathway's mechanisms in determining cell fate.

Associated diseases and disorders

Egr1 is linked to cancer and cardiovascular diseases. Abnormal Egr1 expression and activity have been observed in various cancers where it can act as an oncogene or tumor suppressor depending on the context. Abnormal interactions with proteins such as p53 may contribute to tumorigenesis. In cardiovascular diseases Egr1 influences pathological states by regulating genes associated with inflammation and vascular remodeling. The protein’s dysregulation plays a part in atherosclerosis connecting with inflammatory cytokines to promote disease progression.

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:
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12 product images

Immunohistochemistry (Frozen sections) - Anti-Egr1 antibody [EPR23981-46] (BSA and Azide free) (ab300450)
This data was developed using Anti-Egr1 antibody [EPR23981-46] ab300449, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse liver (fresh) tissue labeling Egr1 with Anti-Egr1 antibody [EPR23981-46] ab300449 at 1/100 (5.48 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Negative control (PMID: 10961876)No staining on mouse liver is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution. .
Immunohistochemistry (Frozen sections) - Anti-Egr1 antibody [EPR23981-46] (BSA and Azide free) (ab300450)
This data was developed using Anti-Egr1 antibody [EPR23981-46] ab300449, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of 4% PFA-fixed, 0.2% Triton X-100 permeabilized frozen Mouse cortex (fresh) tissue labeling Egr1 with Anti-Egr1 antibody [EPR23981-46] ab300449 at 1/100 (5.48 ug/ml) dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution (Green). Positive staining on mouse cortex is observed. The nuclear counterstain was DAPI (Blue). Secondary antibody control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 2 ug/mL dilution. .
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Egr1 antibody [EPR23981-46] (BSA and Azide free) (ab300450)
This data was developed using Anti-Egr1 antibody [EPR23981-46] ab300449, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Mouse kidney tissue labeling Egr1 with Anti-Egr1 antibody [EPR23981-46] ab300449 at 1/2000 (0.274 ug/ml) followed by a ready to use LeicaDS9800 (BOND® Polymer Refine Detection) was used. Negative control: No staining on mouse kidney (PMID: 10961876). The section was incubated with Anti-Egr1 antibody [EPR23981-46] ab300449 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND® Polymer Refine Detection) was used.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Egr1 antibody [EPR23981-46] (BSA and Azide free) (ab300450)
This data was developed using Anti-Egr1 antibody [EPR23981-46] ab300449, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Rat hippocampus tissue labeling Egr1 with Anti-Egr1 antibody [EPR23981-46] ab300449 at 1/2000 (0.274 ug/ml) followed by a ready to use LeicaDS9800 (BOND® Polymer Refine Detection) was used. Positive staining on rat hippocampus. The section was incubated with Anti-Egr1 antibody [EPR23981-46] ab300449 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND® Polymer Refine Detection) was used.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Egr1 antibody [EPR23981-46] (BSA and Azide free) (ab300450)
This data was developed using Anti-Egr1 antibody [EPR23981-46] ab300449, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Mouse hippocampus tissue labeling Egr1 with Anti-Egr1 antibody [EPR23981-46] ab300449 at 1/2000 (0.274 ug/ml) followed by a ready to use LeicaDS9800 (BOND® Polymer Refine Detection) was used. Positive staining on mouse hippocampus. The section was incubated with Anti-Egr1 antibody [EPR23981-46] ab300449 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND® Polymer Refine Detection) was used.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Egr1 antibody [EPR23981-46] (BSA and Azide free) (ab300450)
This data was developed using Anti-Egr1 antibody [EPR23981-46] ab300449, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling Egr1 with Anti-Egr1 antibody [EPR23981-46] ab300449 at 1/500 (1.096 ug/ml) followed by a ready to use LeicaDS9800 (BOND® Polymer Refine Detection) was used. Positive staining on human cerebrum (PMID: 11106242). The section was incubated with Anti-Egr1 antibody [EPR23981-46] ab300449 for 30 mins at room temperature.The immunostaining was performed on a Leica Biosystems BOND® RX instrument Counterstained with Hematoxylin. Secondary antibody only control: Secondary antibody is a ready to use LeicaDS9800 (BOND® Polymer Refine Detection) was used.Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution2) for 20 mins
Immunoprecipitation - Anti-Egr1 antibody [EPR23981-46] (BSA and Azide free) (ab300450)
This data was developed using Anti-Egr1 antibody [EPR23981-46] ab300449, the same antibody clone in a different buffer formulation.

Egr1 was immunoprecipitated from 0.35 mg PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate with Anti-Egr1 antibody [EPR23981-46] ab300449 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-Egr1 antibody [EPR23981-46] ab300449. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/5,000 dilution

Lane 1: PC-12 (rat adrenal gland pheochromocytoma cell) starved overnight, then treated with 100ng/ml NGF and 10uM MG-132 for 2 hours, whole cell lysate 10 μg (Input).
Lane 2: PC-12 starved overnight, then treated with 100ng/ml NGF and 10uM MG-132 for 2 hours, whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Egr1 antibody [EPR23981-46] ab300449 in PC-12 starved overnight, then treated with 100ng/ml NGF and 10uM MG-132 for 2 hours, whole cell lysate (-).

Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

Lysate was freshly made and used immediately to minimize protein degradation.

The bands beneath the target band (80 kDa) are likely to be degraded target fragments.
Lanes 1 - 2: Immunoprecipitation - Anti-Egr1 antibody [EPR23981-46] (Anti-Egr1 antibody [EPR23981-46] ab300449) at 1/30 dilution
Lane 3: Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730)
Lane 1: PC-12 (rat adrenal gland pheochromocytoma cell) starved overnight, then treated with 100ng/ml NGF and 10uM MG-132 for 2 hours, whole cell lysate at 10 µg
Lane 2: PC-12 starved overnight, then treated with 100ng/ml NGF and 10uM MG-132 for 2 hours, whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Egr1 antibody [EPR23981-46] ab300449 in PC-12 starved overnight, then treated with 100ng/ml NGF and 10uM MG-132 for 2 hours, whole cell lysate
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Predicted band size: 57 kDa
Observed band size: 80 kDa
Exposure time: 15s
Immunoprecipitation - Anti-Egr1 antibody [EPR23981-46] (BSA and Azide free) (ab300450)
This data was developed using Anti-Egr1 antibody [EPR23981-46] ab300449, the same antibody clone in a different buffer formulation.

Egr1 was immunoprecipitated from 0.35 mg NIH/3T3 (mouse embryonic fibroblast) whole cell lysate with Anti-Egr1 antibody [EPR23981-46] ab300449 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-Egr1 antibody [EPR23981-46] ab300449. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/5,000 dilution

Lane 1: NIH/3T3 (mouse embryonic fibroblast) starved overnight, then treated with 10ng/ml EGF and 10uM MG-132 for 2 hours, whole cell lysate 10 μg (Input).
Lane 2: NIH/3T3 starved overnight, then treated with 10ng/ml EGF and 10uM MG-132 for 2 hours, whole cell lysate (+).
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Egr1 antibody [EPR23981-46] ab300449 in NIH/3T3 starved overnight, then treated with 10ng/ml EGF and 10uM MG-132 for 2 hours, whole cell lysate (-).

Blocking/Dilution buffer and concentration: 5% NFDM/TBST.

Lysate was freshly made and used immediately to minimize protein degradation.
Lanes 1 - 2: Immunoprecipitation - Anti-Egr1 antibody [EPR23981-46] (Anti-Egr1 antibody [EPR23981-46] ab300449) at 1/30 dilution
Lane 3: Immunoprecipitation - Rabbit IgG, monoclonal [EPR25A] - Isotype Control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730)
Lane 1: NIH/3T3 (mouse embryonic fibroblast) starved overnight, then treated with 10ng/ml EGF and 10uM MG-132 for 2 hours, whole cell lysate at 10 µg
Lane 2: NIH/3T3 starved overnight, then treated with 10ng/ml EGF and 10uM MG-132 for 2 hours, whole cell lysate
Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Egr1 antibody [EPR23981-46] ab300449 in NIH/3T3 starved overnight, then treated with 10ng/ml EGF and 10uM MG-132 for 2 hours, whole cell lysate
Secondary
All lanes: Immunoprecipitation - VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366) at 1/5000 dilution
Predicted band size: 57 kDa
Observed band size: 80 kDa
Exposure time: 24s
Immunocytochemistry/ Immunofluorescence - Anti-Egr1 antibody [EPR23981-46] (BSA and Azide free) (ab300450)
This data was developed using Anti-Egr1 antibody [EPR23981-46] ab300449, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized PC-12 (rat adrenal gland pheochromocytoma cell) cells labelling Egr1 with Anti-Egr1 antibody [EPR23981-46] ab300449 at 1/100 dilution (5.48 μg/ml), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing mainly nuclear staining in PC-12 cells, which were first starved for 18 hours, then treated with EGF (10 ng/ml) and MG-132(10 uM) for 2 h. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used as counterstain at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
Immunocytochemistry/ Immunofluorescence - Anti-Egr1 antibody [EPR23981-46] (BSA and Azide free) (ab300450)
This data was developed using Anti-Egr1 antibody [EPR23981-46] ab300449, the same antibody clone in a different buffer formulation.

Immunofluorescent analysis of 4% Paraformaldehyde-fixed, 0.1% TritonX-100 permeabilized NIH/3T3 (mouse embryonic fibroblast) cells labelling Egr1 with Anti-Egr1 antibody [EPR23981-46] ab300449 at 1/100 dilution (5.48 μg/ml), followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed antibody at 1/1000 dilution (Green).

Confocal image showing mainly nuclear staining in NIH/3T3 cells, which were first starved for 18 hours, then treated with EGF (10 ng/ml) and MG-132(10 uM) for 2 h. Image was taken with a confocal microscope(Leica-Microsystems, TCS SP8).

Alexa Fluor® 594 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker ab195889 Anti-alpha Tubulin mouse monoclonal antibody - Microtubule Marker (Alexa Fluor® 594) was used as counterstain at 1/200 dilution (Red). The Nuclear counterstain was DAPI (Blue).

Secondary antibody only control: Secondary antibody is Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed ab150081 Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution.
Western blot - Anti-Egr1 antibody [EPR23981-46] (BSA and Azide free) (ab300450)
This data was developed using Anti-Egr1 antibody [EPR23981-46] ab300449, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used as loading control for GAPDH.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.
All lanes: Western blot - Anti-Egr1 antibody [EPR23981-46] (Anti-Egr1 antibody [EPR23981-46] ab300449) at 1/1000 dilution
Lane 1: Untreated PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg
Lane 2: PC-12 starved overnight, then treated with 100ng/ml NGF and 10uM MG-132 for 2 hours whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 57 kDa
Observed band size: 80 kDa
Exposure time: 15s
Western blot - Anti-Egr1 antibody [EPR23981-46] (BSA and Azide free) (ab300450)
This data was developed using Anti-Egr1 antibody [EPR23981-46] ab300449, the same antibody clone in a different buffer formulation.

Blocking and diluting buffer and concentration: 5% NFDM/TBST.

Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 was used for GAPDH loading control.

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

The identity of the lower MW band at approximately 20 kDa is unknown.
All lanes: Western blot - Anti-Egr1 antibody [EPR23981-46] (Anti-Egr1 antibody [EPR23981-46] ab300449) at 1/1000 dilution
Lane 1: Untreated THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg
Lane 2: THP-1 treated with 100ng/ml LPS (lipopolysaccharide) for 3 hours whole cell lysate at 20 µg
Lane 3: Untreated NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg
Lane 4: NIH/3T3 starved overnight, then treated with 10ng/ml EGF and 10uM MG-132 for 2 hours whole cell lysate at 20 µg
Secondary
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/20000 dilution
Predicted band size: 57 kDa
Observed band size: 80 kDa
Exposure time: 180s